Māori and Pacific Islander people are a priority population originating from Australasia. Māori and Pacific Islander children exhibit greater risk of obesity and associated morbidities compared to children of other descent, secondary to unique cultural practices and socioeconomic disadvantage. Despite these known risk factors, there is limited synthesised evidence for preventing and treating childhood obesity in this unique population. The objective of this systematic review was to identify and evaluate global prevention or treatment interventions for overweight or obesity that targeted Māori and Pacific Islander children and adolescents (aged 2–17 years).The Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines were followed. The databases PubMed, EMBASE, Scopus, Web of Science and CINAHL were searched from inception to August 2018. Study quality and risk of bias was assessed using a modified Downs and Black Quality Checklist for Health Care Intervention Studies. Studies were included if RCT/intervention/case control/ or prevention study designs. The study group was defined under the search term ‘Oceanic Ancestry Group’.Of the initial 94 articles identified, six were included describing two prevention and three treatment interventions. Interventions were heterogenous in setting, design, length and outcomes. Four interventions were implemented in New Zealand. Most studies were of ‘fair’ quality. One study recruited an exclusive population of Māori and Pacific Islander participants. In the five studies that recruited mixed populations, one performed sub-group analysis on Māori and Pacific Islander participants. No study reported an improvement in anthropometric outcomes post-intervention in complete or sub-group analysis. Improvements in cardiometabolic or psychological secondary outcomes were inconsistent across all studies.There is a lack of evidence to recommend specific intervention characteristics to optimise obesity prevention or treatment outcomes for Māori and Pacific Islander children. Future research requires greater consideration of cultural values and beliefs, community engagement, exclusive targeting of Māori and Pacific Islander children and families, and sub-group analyses for mixed-population studies. Incorporating co-design principles during study design and implementation can maximise the cultural specificity of interventions and may contribute to improved health and weight-related outcomes for this at-risk, priority population.CRD42019121790 (26 March 2019).PROSPERO  The authors concluded that a co-design approach empowered the target population by enabling active, participatory action throughout each phase of design [In their recently published article, Verbiest et al.  describef design .Whilst standalone trials are essential in building an evidence-base for appropriate childhood overweight/obesity prevention and intervention characteristics in Māori and Pacific Islander peoples, a coordinated whole-of-government, policy-level and healthcare systems response is paramount in contributing to a reduction in prevalence. In response to the growing prevalence of obesity and congruent with the World Health Organization (WHO) Report of the Commission on Ending Childhood Obesity , New ZeaA localised example of a culturally-tailored, integrated health system approach to improving Māori and Pacific Islander child and adolescent health is the ‘Good Start Program’, a statewide, schools-based, nutrition and physical activity initiative embedded within the Queensland Health system, Australia. A qualitative evaluation of its implementation concluded that building a system-wide workforce of Māori and Pacific Islander Health Workers was a significant contributor to enabling high-level community engagement, cultural specificity of interventions, community member satisfaction and trust with the Good Start Program, and ultimately positively shifting Māori and Pacific Islander community members’ attitudes to and knowledge of health .There were a number of observed strengths of included studies. Project Energize, the New Zealand schools-based prevention intervention described by Rush et al. , 26 in tn = 18) and the participants of SWITCH trial of Maddison et al. [n = 116), which may have limited the generalisability of findings to the larger population of Hawaii. Most interventions were described as stand-alone, and there was little evidence of attempted integration within a broader, health system to maximise reach, impact, and sustainability.The primary limitation of the current literature is the lack of exclusive recruitment of Māori and Pacific Islander participants and limited sub-group analysis in mixed participant demographics. Descriptions of intervention intensity and intervention fidelity could have been strengthened; this may have contributed to the modesty of reported results, especially anthropometric outcomes. The intervention of Chansavang et al.  was impln et al.  were offn et al. . Anothern et al. , 25–27 an et al.  and 5-yen et al.  analysedThe primary strength of this review is its novelty, being the first known to synthesise global evidence of prevention and treatment interventions for the unique, at-risk population of Māori and Pacific Islander children and adolescents. Findings of this review are potentially transferable to priority populations from various developed countries, including the USA and Canada, where young Alaska Native and American Indian children, and First Nations and Inuit peoples, respectively, exhibit high obesity prevalence rates . As an en = 6) to prevent or treat obesity in Māori and Pacific Islander children and adolescents generated minimal impact in improving anthropometric indicators of weight or improving cardiometabolic or psychological secondary outcomes. There is a lack of evidence to recommend specific intervention characteristics that will optimise overweight/obesity prevention or treatment interventions in Māori and Pacific Islander children and adolescents. These results are possibly secondary to a lack of intervention intensity and specificity to Māori and Pacific Islander peoples. The authors propose the following recommendations for future research:Cultural-tailoring of interventions, preferably utilising a co-design approach, with adequate methodological reporting;Implementation of interventions that exclusively target Māori and Pacific Islander children and adolescents; fostering community engagement, leadership and ownership at every stage of the proposed intervention i.e. from conception to evaluation;Performing intervention sub-group analysis on Māori and Pacific Islander participants in mixed-population studies; andIntegrating and evaluating, where possible, long-term, mixed-methods interventions within an existing healthcare system to maximise reach and sustainability for policy- and population-level impactOverall, previously reported studies (Consideration of these recommendations in future research will optimise interventions to tackle childhood overweight/obesity in the unique priority population of Māori and Pacific Islanders, who exhibit a significantly higher prevalence of childhood overweight/obesity in Australia and New Zealand, as well as demonstrate substantial socioeconomic and health disadvantage that inherently increases population risk for long-term overweight/obesity and its comorbidities.Additional file 1. PRISMA checklist.docxAdditional file 2. Search strategy in EMBASE according to the PICO format.docxAdditional file 3. Individual quality assessment and risk of bias results for included studies.docx


Cannabis Sativa plant, interacts with the endocannabinoid system by inhibiting fatty acid amide hydrolase (FAAH) activity (the rate-limiting enzyme for anandamide hydrolysis), allosterically modulating CB1 and CB2 receptors, and activating components of the “extended endocannabinoid system.” Congruently, CBD has shown prominent pro-neurogenic effects, and, unlike Δ9-tetrahydrocannabinol, it has the advantage of being devoid of psychotomimetic effects. Here, we first review pre-clinical studies supporting the facilitating effects of CBD on adult hippocampal neurogenesis and available data disclosing cannabinoid mechanisms by which CBD can induce neural proliferation and differentiation. We then review the respective implications for its neuroprotective, anxiolytic, anti-depressant, and anti-reward actions. In conclusion, accumulating evidence reveals that, in rodents, adult neurogenesis is key to understand the behavioral manifestation of symptomatology related to different mental disorders. Hence, understanding how CBD promotes adult neurogenesis in rodents could shed light upon translational therapeutic strategies aimed to ameliorate psychiatric symptomatology dependent on hippocampal function in humans.During the last decades, researchers have investigated the functional relevance of adult hippocampal neurogenesis in normal brain function as well as in the pathogenesis of diverse psychiatric conditions. Although the underlying mechanisms of newborn neuron differentiation and circuit integration have yet to be fully elucidated, considerable evidence suggests that the endocannabinoid system plays a pivotal role throughout the processes of adult neurogenesis. Thus, synthetic, and natural cannabinoid compounds targeting the endocannabinoid system have been utilized to modulate the proliferation and survival of neural progenitor cells and immature neurons. Cannabidiol (CBD), a constituent of the  During the last half-century, considerable progress has been made to understand, prevent, and treat such conditions. However, treatment options are still far from optimal in terms of efficacy and specificity, and there remain important untreatable maladaptive phenotypes and treatment-resistant patients. To solve this issue, basic and applied research has tried to identify new altered neuropsychological mechanisms suitable to promote new therapeutic strategies , CBD is devoid of psychotomimetic and rewarding effects , transient potential vanilloid 1 (TRPV1), G-protein 55 (GPR55) and peroxisome proliferator-activated gamma (PPARγ) receptors, as well as the antagonism of adenosine reuptake  of the dentate gyrus . Moreover, the authors reported an interesting opposition to the effects of THC on this measure. Months later, Demirakca et al. (Considering that the endocannabinoid (eCB) system exerts important functions in the regulation of neuronal generation and survival  and BrdU/NeuN. Due to the prolonged presence of both markers in different stages of the neurogenesis process (for a review see Kempermann et al., in vitro, Campos et al. (in vivo. Inverted U-shaped dose-response curves usually suggest the participation of multiple pharmacological mechanisms. In this way, it has already been described that CBD also exerts a similar anxiolytic dose-response curve (for a review see Jurkus et al., 1A and TRPV1 mechanisms (Campos and Guimarães, 1A receptors. However, CBD-induced proliferation in HiB5 hippocampal progenitor cells was not blocked by a 5-HT1A antagonist (Campos et al., CBD pro-neurogenesis also shows great consistency across doses. Literature findings report increases in neuronal proliferation and differentiation after CBD doses ranging from 3 to 30 mg/kg, usually after prolonged treatments (≥10 days; s et al.  describes et al.  showed t−/− mice (Aguado et al., The eCB system stands out as a key regulator of newborn neuron generation, survival, maturation, and functional integration in the adult hippocampus. Neural progenitor cells, and their descendants, express a functional eCB system and are subject to the effects of endocannabinoid signaling (Prenderville et al., in vitro and in vivo evidence has suggested such interplay. The first evidence was given by Wolf et al. (−/− mice. The seminal work of Campos et al. (in vitro. CB1 and CB2 antagonists prevented the pro-neurogenic effect of CBD in hippocampal HiB5 progenitor cells. Furthermore, CB1 and CB2 receptor agonists, as well as eCB degradation inhibitors mimicked the pro-neurogenic effects of CBD. Interestingly, CBD effects were abrogated when the FAAH was inhibited. Combined, these results imply that the pro-neurogenic effects of CBD depending on the increase of AEA concentration. Crucially, CBD is an inhibitor of the FAAH and is well known to increase AEA concentration (Bisogno et al., in vivo. After a CBD treatment in chronically stressed mice, neuronal differentiation, and late survival were found to be increased in CBD-treated mice (Fogaça et al., Given the mechanistic interactions between CBD and eCB system, a plausible hypothesis originated stating that CBD increases adult neurogenesis by modulating the eCB system. Accordingly, f et al. . In theis et al.  further A considerable number of studies have reported the pro-neurogenic effects of CBD, and some among them have even related these with an eCB mechanism of action. But, can the pro-neurogenic effects of CBD account for some of its therapeutic applications? Answering this question requires specialized experimental strategies designed to rule out CBD pro-neurogenesis, leaving intact its other pharmacological mechanisms and so, fewer experiments have been conducted. Nonetheless, a handful of studies have addressed this question, presenting evidence for a potential implication in the protection against neurodegenerative diseases (Esposito et al., Neurodegenerative and ischemic conditions are among the circumstances in which hippocampal function can manifest greater impairments (Shah et al., A significant amount of animal and human data has emerged relating the neuro-modulatory role of adult hippocampal neurogenesis, its interactions with broader hippocampal circuits, and its implications on altered behaviors in different neuropsychiatric disorders (Beckervordersandforth and Rolando, ML and OV were responsible for the study concept and design. Both authors drafted the manuscript and approved the final version for publication.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Cell death (CD) is a fundamental biological process that is indispensable in all living organisms Ameisen, . Phloem + outward rectifying conductances (KORC) was recorded in response to various CD-inducing microbe-derived molecules, such as harpins , cell shrinkage (Yekkour et al., + efflux channel were also shown to have fewer autophagosomes compared to the wild-type plant upon ROS-induced CD (Demidchik, + efflux through KORC is supposed to result in dramatic K+ loss from plant cells and promotes CD (Demidchik et al., + loss was effectively shown to be involved in tobacco cell death induced by palmitoleic acid and ceramide (Peters and Chin, Nicotiana benthamiana plants undergoing oxidative stress and transiently expressing CED-9, an anti-apoptotic gene from the bcl-2 family (Craig, + efflux and maintaining intracellular K+ homeostasis (Shabala et al., An increase in Ky Craig, , are capIn response to ozone or DON, plant cells rapidly activate anion currents, followed by a delayed activation of KORC (Tran et al., In plant the vacuolar collapse seems a key step in cell shrinkage but to our knowledge the role of vacuolar conductances is poorly documented. Therefore, further studies are needed to decipher the role of vacuolar channels in this process.+ channels and is prevented by application of blockers of Cl− or K+ channels (Okada and Maeno, As a whole, these data are reminiscent of those described in numerous studies in animal cells and show that plant CD could involve specific modifications of ion transporter activities that could be significant and crucial in the successful propagation of CD . ActivatFB, DR, DT, and PL conceived and wrote the manuscript. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Anorexia nervosa and bulimia nervosa are two severe eating disorders associated with high premature mortality, suicidal risk and serious medical complications. Transition between anorexia nervosa and bulimia nervosa over the illness course and familial co-aggregation of the two eating disorders imply aetiological overlap. However, genetic and environmental liabilities to the overlap are poorly understood. Quantitative genetic research using clinical diagnosis is needed.N = 782 938) of randomly selected full-sisters and maternal half-sisters born in Sweden between 1970 and 2005. Structural equation modelling was applied to quantify heritability of clinically diagnosed anorexia nervosa and bulimia nervosa and the contributions of genetic and environmental effects on their overlap.We acquired a clinical diagnosis of anorexia nervosa  and bulimia nervosa  in a large population-based sample  (36–50%)] and 41% (31–52%), respectively, in the study population, with the remaining variance explained by variance in unique environmental effects. We found statistically significant genetic  and unique environmental correlations [0.55 (0.43–0.66)] between the two clinically diagnosed eating disorders; and their overlap was about equally explained by genetic and unique environmental effects [co-heritability 47% (35–58%)].Our study supports shared mechanisms for anorexia nervosa and bulimia nervosa and extends the literature from self-reported behavioural measures to clinical diagnosis. The findings encourage future molecular genetic research on both eating disorders and emphasize clinical vigilance for symptom fluctuation between them. Anorexia nervosa is characterized by significantly low body weight and intense fear of weight gain  of bulimia nervosa compared to the same types of relatives of individuals without anorexia nervosa, and In the current work, we used clinical diagnoses of anorexia nervosa and bulimia nervosa in a large Swedish female cohort containing full-sisters  and maternal half-sisters . The application of diagnostic information considerably expands the clinical significance of the results. Using non-twin siblings significantly increased sample size and improved precision in estimating the underlying morbidity liabilities. We quantified the contribution of genetic and environmental effects on anorexia nervosa and bulimia nervosa and their overlap and estimated the genetic and environmental correlations between the two eating disorders.The current study was approved by the Regional Ethics Review Board in Stockholm, Sweden.et al., et al., et al., et al., et al., We acquired information from several Swedish national registers linked by the unique individual identification number. The Swedish Population Register provided birth year and month, death date and migration information  from the treatment quality registers. We were unable to distinguish between restricting and binge-eating and purging anorexia nervosa from the ICD diagnosis in the NPR. Bulimia nervosa diagnosis was identified with ICD-10 codes F50.2 or F50.3 from the NPR, or meeting DSM-IV-TR criteria for bulimia nervosa or sub-threshold bulimia nervosa in EDNOS from the treatment quality registers. Bulimia nervosa was not an independent eating disorder category in the Swedish version of ICD-9 . Therefos.d.) = 9.3]. We then randomly selected one pair of full-sisters per mother in the remaining mothers after excluding mothers of the selected maternal half-sisters. We excluded twin pairs from full-sister pairs, resulting in 334 433 pairs of full-sisters for analysis . No individual was included in more than one pair. The selected full-sisters and maternal half-sisters comprised the study population, where 6104 (0.91%) individuals among the full-sisters and 938 (0.82%) among the maternal half-sisters had been diagnosed with anorexia nervosa; 3142 (0.47%) among the full-sisters and 579 (0.51%) among the maternal half-sisters had been diagnosed with bulimia nervosa; 679 (0.10%) among the full-sisters and 122 (0.11%) among the half-sisters had been diagnosed with both disorders  and maternal half-sisters (sisters born to the same mother but different fathers) who were born in Sweden between 1970 and 2005. We excluded individuals who emigrated or died before age 6, adopted individuals and individuals whose biological parents were not identifiable from the registers. We did not have access to information on race and ethnicity; however, being born in Sweden might suggest that our study population was comprised primarily of individuals of Scandinavian/Nordic ancestry. We then randomly selected one pair of maternal half-sisters per mother  in full-sisters and 0.03 (s.e. = 0.06) in maternal half-sisters; the cross-sister-within-trait correlation for bulimia nervosa was estimated as 0.20 (s.e. = 0.03) in full-sisters and 0.13 (s.e. = 0.07) in maternal half-sisters . The AE model was selected for interpretation because it had the lowest AIC , illustrating the diagnostic transition between the two eating disorders within individuals regardless of the type of siblings. The cross-sister-cross-trait correlation further characterizes the nature of the association between the disorders across the individuals in a pair. A higher cross-sister-cross-trait correlation was observed in full-sister pairs  than in maternal half-sisters , suggesting genetic effects on the overlap between the two eating disorders ] of the phenotypic correlation of the two disorders was explained by their genetic covariance and the remaining 54% [95% CI (42–65%)] was explained by their environmental covariance . We founet al., s.e. 1%) in the most updated genome-wide association study (GWAS) ] and environmental correlations [0.55 (0.43–0.66)] suggest an aetiological overlap between anorexia nervosa and bulimia nervosa and underscore the importance of vigilance for transitions between the two eating disorders during treatment. Using large population data, we significantly improved the precision of the estimates. We reported heritability for clinically diagnosed anorexia nervosa as 43% (36–50%) and bulimia nervosa as 41% (31–52%), which was slightly lower than, but within the confidence interval of, the heritability found in the twin study based on self-reported questionnaire data [57% (0–81%) for anorexia nervosa and 62% (8–70%) for bulimia nervosa] ] and between anorexia nervosa and major depression (around 0.5 in multiple studies) (Wade et al., et al., et al., et al., In addition, we reported significant unique environmental effects, both disorder-specific and common to the two eating disorders. Previous research suggested various potential environmental risk factors for eating disorders, such as dieting and events that trigger mood change and thin-ideal internalization (Chua et al., et al., et al., Using extended familial relatedness drastically increased the available sample size of the models in our study, and the use of clinical diagnostic information increased the clinical significance of the results. Our results are consistent with the prior twin study on the overlap between anorexia nervosa and bulimia nervosa based on self-reported measures of eating behaviours (Bulik et al., et al., et al., et al., et al., et al., et al., et al., et al., et al., et al., et al., Limitations of the current study would arise if there are violations of basic assumptions of the models, such as the assumption of equal shared environmental factors across different types of siblings [equal environment assumption, EEA (Plomin To our knowledge, this is the first quantitative genetic study on clinically diagnosed anorexia nervosa and bulimia nervosa and their overlap. The moderate and statistically significant genetic and environmental correlations between the two eating disorders might suggest aetiological overlaps and emphasize clinical vigilance of the transition between the two disorders.


In modern biotechnological and medical research, RNA-guided nucleases (RGNs) continue to be highly effective in targeted modification of genomes and the manipulation of gene expression (Sander and Joung CRISPR-Cas proteins are non-specific endonucleases that bind a protospacer adjacent motif (PAM) located in the proximity of the genomic target  nuclease concentration and features and (ii) target site accessibility, functionality, and uniqueness. The occurrence of complementary sites within the genome, which form highly stable duplexes to guide oligonucleotide, is one of the important determinants of off-target activity. Cas9 and Cas12a (Cpf1) activity can be modulated by chromatin states to varying degrees. Off-targeting is expected to be context dependent because chromatin states and DNA accessibility is tissue/cell- or condition-dependent  RISC complex formation, several key measurements including association kinetics, equilibrium binding energies, and single turnover cleavage rates allows revealing of important rules for binding and cleavage of targets. Becker and co-authors (Becker et al. A comparison of Cas9- and Cas12a-binding experiments showed variable binding kinetics responses to target sequence mutations, which explained why Cas12a enables the selection of DNA sequences more precisely than Cas9 (Strohkendl et al. For Cas nucleases, targeting rules were empirically established by different groups (Klein et al. Several models and approaches based upon thermodynamics and kinetics have the potential to explain off-targeting patterns for CRISPR-Cas and AGO2, as well as for other systems. To go beyond binding energetics, Klein and co-authors (Klein et al. Bisaria and co-authors (Bisaria et al. Attenuating DNA cleavage kinetics can be successfully applied for enhancing gene editing specificity and reducing off-targeting not only to CRISPR systems but also to different engineered nucleases (Becker et al.


Ichihashi et al reported that 43% of patients had antipsychotic polypharmacy. Number of antipsychotics used in patients with schizophrenia in Japan was the greatest among Asian countries. However, the antipsychotic polypharmacy rate in Japan decreased gradually. Recent systematic review, meta‐analysis and meta‐regression analysis demonstrated that antipsychotic augmentation was superior to monotherapy. However, several cohort studies have suggested a significant association between antipsychotic daily dose and mortality. In addition, most pharmacokinetic interactions with antipsychotics occur at the metabolic level and usually involve changes in the activity of the major drug‐metabolizing enzymes involved in their biotransformation. Thus, avoidance of unnecessary polypharmacy, knowledge of the interaction profiles of individual agents, and careful individualization of dosage based on close evaluation of clinical response and possibly plasma drug concentrations are essential to prevent and minimize potentially adverse drug interactions in patients receiving antipsychotics. Ichihashi et alAntipsychotics are not always effective in schizophrenia patients. There are still patients with treatment‐resistant schizophrenia (TRS), which has led to a growing trend of resorting to atypical antipsychotic polypharmacy due to fewer side effects.Number of antipsychotics used and psychotropic drug loading in patients with schizophrenia in Japan was the greatest among Asian countries.A recent systematic review, meta‐analysis and meta‐regression analysis demonstrated that antipsychotic augmentation was superior to monotherapy regarding total symptom reduction, although the superiority was only apparent in open‐label and low‐quality trials and not in double‐blind and high‐quality trials.On the other hand, several guidelines consistently recommend monotherapy of antipsychotic agents. For example, the Maudsley Prescribing Guidelines in Psychiatry states that we should concern ourselves with the addition of an antipsychotic to another antipsychotic solely to increase efficacy. From a theoretical point of view, since all antipsychotics block D2 receptors , there is limited rationale for additional antipsychotics. In addition, many clinical trials studying the side effects of polypharmacy have associated polypharmacy with an increased incidence of side effects, although most have been uncontrolled studies or observational studies.Several cohort studies have suggested a significant association between antipsychotic daily dose and mortality.Mortality associated with antipsychotic polypharmacy has not yet been concluded. Previous reports have suggested that significant mortality risks are 2.46 Thus, although evident disadvantage of antipsychotic polypharmacy has not yet been concluded, avoidance of unnecessary polypharmacy, knowledge of the interaction profiles of individual agents, and careful individualization of dosage based on close evaluation of clinical response and possibly plasma drug concentrations are essential to prevent and minimize potentially adverse drug interactions in patients receiving antipsychotics.Norio Yasui‐Furukori has been a speaker for Otsuka Pharmaceutical Co., Ltd., Mochida Pharmaceutical Co., Ltd., Dainippon‐Sumitomo Pharmaceutical Co., and MSD Co. Kazutaka Shimoda has received research support from Novartis Pharma KK, Dainippon Sumitomo Pharma Co., Astellas Pharma Inc, Meiji Seika Pharma Co., Ltd., Eisai Co., Ltd., Pfizer Inc, Otsuka Pharmaceutical Co., Ltd., Daiichi Sankyo Co., and Takeda Pharmaceutical Co., Ltd., and honoraria from Eisai Co., Ltd., Mitsubishi Tanabe Pharma Corporation, Takeda Pharmaceutical Co., Ltd., Meiji Seika Pharma Co., Ltd., Janssen Pharmaceutical KK, Shionogi & Co., Ltd., Dainippon Sumitomo Pharma Co., Daiichi Sankyo Co., and Pfizer Inc The funders did not have any role in data collection or in the study design, analysis, decision to publish, or preparation of the manuscript. The remaining authors declare that they have no competing interests to report.NYF designed the study and wrote the initial draft of the manuscript. KS contributed to supervision of the draft.


On page 288, ‘recent reports have shown that CAII can also reduce nitrite (NO2−) to nitric oxide (NO), and thus, may play a role in vasodilation and regulation of blood pressure  to nitric oxide (NO), and thus, may play a role in vasodilation and regulation of blood pressure  On pages 288–299, ‘However, when dialyzed with ethylenediaminetetraacetic acid (EDTA), the enzyme retained its carbonic anhydrase activity yet lost its nitrite reductase activity  And on page 291, ‘... indicating that a metal cofactor within the bovine blood was needed for the CAII-dependent nitrite reductase activity (Andring et al., 2018et al., 2018et al. (2018et al., 2018( al. 2018 and shou


A discussion of recent discoveries in the etiolation/de-etiolation field, focusing on post-transcriptional processes and ultrastructural changes, along with comments on usage of the term ‘etiolation’, and of common etiolation/de-etiolation systems. The state of etiolation is generally defined by the presence of non-green plastids (etioplasts) in plant tissues that would normally contain chloroplasts. In the commonly used dark-grown seedling system, etiolation is coupled with a type of growth called skotomorphogenesis. Upon illumination, de-etiolation occurs, marked by the transition from etioplast to chloroplast, and, at the seedling level, a switch to photomorphogenic growth. Etiolation and de-etiolation systems are therefore important for understanding both the acquisition of photosynthetic capacity during chloroplast biogenesis and plant responses to light—the most relevant signal in the life and growth of the organism. In this review, we discuss recent discoveries (within the past 2–3 years) in the field of etiolation and de-etiolation, with a particular focus on post-transcriptional processes and ultrastructural changes. We further discuss ambiguities in definitions of the term ‘etiolation’, and benefits and biases of common etiolation/de-etiolation systems. Finally, we raise several open questions and future research possibilities. Etiolation involves prolonged growth in the absence of light that results in the development of etioplasts in tissue that would have chloroplasts if subjected to light. Etioplasts do not contain chlorophyll or stacked thylakoid membranes, but rather have a paracrystalline lipid–pigment–protein structure known as the prolamellar body (PLB). The PLB consists largely of the plastid lipids monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG), and an association of the chlorophyll precursor protochlorophyllide (Pchlide), the light-dependent protochlorophyllide oxidoreductase (LPOR) that is responsible for its conversion, and the cofactor NADPH  germination and growth of seedlings in complete darkness, the term ‘etiolated’ is commonly defined additionally by the presence of a skotomorphogenic phenotype of elongated hypocotyls, shortened roots, and small, closed cotyledons . In thesThe majority of the data discussed in this Expert View refer to work undertaken in such seedling-based etiolation/de-etiolation systems. The various limitation of these systems and possible alternative or complementary systems are also discussed (in the section ‘New systems required and new lessons learned’).étiolier (i.e. straw), is still used as a descriptor for a range of pale or yellowing phenotypes. These include nitrogen-deficient rice  albostrians mutant  are 20–24 nt-long molecules that regulate gene expression via RNA-dependent DNA methylation, translation inhibition, or mRNA cleavage (reviewed in Zea mays) to understand the establishment of photosynthesis in C3 versus C4 species. These studies, which defined several specific sRNA roles, such as the repression of photomorphogenic growth by miR396 via members of the Growth Regulating Factors family  is an evolutionarily conserved protein kinase that acts as a central hub to control cellular- and organism-level development : RNA–protein complexes that are conserved in eukaryotes and regulate gene expression by degradation or translational arrest of mRNA (reviewed in Genomes uncoupled (gun) mutants defective in plastid-to-nucleus retrograde signaling (gun2–6) have defects in genes for enzymes involved in tetrapyrrole biosynthesis. More recently, a role for the enigmatic GUN1 in regulating protein import via the cytosolic heat shock protein 90 (HSP90) chaperone was clarified using a de-etiolation system  is produced early during greening as a by-product of tetrapyrrole biosynthesis  and lipid to protein ratios change with greening . The rolin vitro system recently used to show the requirement for MGDG and charged lipids in regulating LPOR complex formation and activity . These issues argue for alternative systems, such as the ues e.g. .Phaseolus coccineus) and pea (Pisum sativum) (Landoltia punctate), a flat-leafed aquatic monocot  requires limited growth time and space yet provides sufficient material compared with other experimental systems such as the shoot apical meristem, and (ii) is highly customizable by use of different timing and lighting regimes and introduction of different substances to the growth medium (sativum) . PLBs hasativum) . The pro monocot , and in Cucumis sativus; Persea americana; Phaseolus vulgaris) (Nicotiana tabacum)  thylakoid membranes, early studies in cucumber (ulgaris) , varioustabacum) . These fThe benefits of the standard seedling etiolation and/or de-etiolation systems means that they have been used often in recent years to study diverse topics including gravitropism  changes during greening in Arabidopsis , acts downstream of the COP1–auxin cascade during de-etiolation  in the dark . Soluble1O2; 1O2 retrograde signaling mediates de-etiolation via the EXECUTER1 pathway (The early assembly of the PSII oxygen evolving complex results in the (damaging) formation of singlet oxygen (Three recent studies investigated the effect of decreased MGDG (


Recent years have seen the idea of a close association between nutrition and the modulation of cancer development/progression reinforced. In fact, an increasing number of experimental and epidemiological evidence has been produced, supporting the concept that many different bioactive components of food  may be implicated in either the promotion of or the protection against carcinogenesis. At the cellular level, such compounds can have an impact on different but sometimes intertwined processes, such as growth and differentiation, DNA repair, programmed cell death, and oxidative stress. In addition, compelling evidence is starting to build up of the existence of primary epigenetic targets of dietary compounds, such as oncogenic/oncosuppressor miRNAs or DNA-modifying enzymes, which in turn impair gene expression and function. This editorial aims to summarize the themes of the 31 papers  published in the Special Issue “Role of Natural Bioactive Compounds in the Rise and Fall of Cancers” presenting the latest findings on the intracellular pathways and mechanisms affected by selected natural molecules influencing the fine-tuning of cancer phenotype.Plant polyphenols have been among the most studied natural compounds by the contributors to this issue.In the original article group, Polonio-Alcalà et al.  showed tOther natural phenolic compounds whose activity is discussed in the original articles of this issue are:–Gallic acid , widely distributed in natural plants, fruits, and green tea, whose tumor-suppressive effect via the p53-mediated downregulation of the transmembrane protein PD-L1 was demonstrated by Kang et al. [–Olea europaea L., whose presence in enriched extracts from olive leaves was proven to reduce the glycolytic rate of a wide range of solid and liquid tumor cells via the downregulation of the three key effectors of the glycolytic pathway, GLUT-1, PKM2 and MCT4, likely resulting in a decreased glucose entrance and biomass production [Oleuropein, the main bioactive phenolic component of oduction ;–Oleacein , the main secoiridoid contained in extra virgin olive oil, able to elicit significant anti-tumor activity by promoting cell cycle arrest and apoptosis in multiple myeloma cells due to its histone deacetylase inhibitory properties ;–Dadzein , present in soybeans, whose 4-sulphate metabolite produced by gut microbiota was found to exert an anti-estrogenic effect on ERα-positive breast cancer cells via the downregulation of the anti-apoptotic neuroglobin protein thus rendering cells more prone to the paclitaxel treatment [–Gigantol, a bibenzyl compound from orchid species, whose ability to destabilize tumor integrity via the suppression of the PI3K/AKT/mTOR and JAK/STAT pathways was demonstrated by Losuwannarak et al.  in non-s–Lonchocarpus sericeus, proven to be a powerful inhibitor of the Wnt/β-catenin pathway able to selectively suppress the migration and proliferation of a panel of colorectal cancer cell lines in vitro and in a preclinical colorectal cancer mouse model [Lonchocarpin, a chalcone isolated from se model ;–Isorhamnetin, , a flavonol aglycone found in some medicinal plants, able to exert an anti-proliferative effect on human bladder cancer cells via the induction of cell cycle arrest during the G2/M phase and apoptosis, accompanied by the activation of the AMPK signaling pathway and ROS overproduction ;–Piper genus plants, whose ability to inhibit XIAP protein, involved in the regulation of caspase-dependent/independent cell death pathways, was reported by Muñoz et al. [Erioquinol, eriopodol A and gibbilimbol B, derived from z et al.  in breas–Calocedrus formosana Florin leaves extract, proven to interfere with cell cycle and microtubule dynamics in lung adenocarcinoma cells, also inhibiting tumor growth in a xenograft mouse model [Vatein, isolated from se model .Magnolia-derived polyphenol honokiol based upon its ability to impair cell cycle progression, inhibit epithelial–mesenchymal transition, and suppress cell motility, invasion, metastasis and angiogenesis. Zhou et al. [In the Review section, Perrone et al.  discusseu et al.  summarizu et al.  reviewedu et al.  focused –Malva pseudolavatera leaves, which showed a promising selective anti-proliferative and pro-apoptotic effect on acute myeloid leukemia cell lines, determining PARP cleavage, cytochrome-c release, Bax/Bcl-2 ratio increase and ROS overproduction [The methanolic extract of oduction ;–Eicosapentaenoic acid, an ω-3 polyunsaturated fatty acid, which played a protective role, both alone and in combination with angiotensin-converting enzyme inhibitors, in attenuating adipocyte-induced proinflammatory cytokine expression and the migration of breast cancer cells in an in vitro model of obesity-induced breast cancer ;–Fucoidan, a sulphated polysaccharide derived from brown seaweed, whose combination with gemcitabine determined an enhanced pro-apoptotic and cell cycle-inhibitory activity on selected uterine carcinosarcoma and stromal sarcoma cell lines ;–Nicotin, whose mechanisms underlying the promotion of melanoma cell proliferation and migration mediated through α9-nAChR-initiated carcinogenic signaling and PD-L1 expression were reported by Nguyen et al. ;–Streptomyces sp. MUM256, isolated from mangrove soil in Malaysia, and the cyclic dipeptides contained whose ability to induce cell cycle arrest and apoptosis was demonstrated by Tan et al. [The ethyl acetate fraction of the crude extract of n et al.  in colon–Luffariella variabilis, which preferentially inhibits the proliferation of oral cancer cells inducing apoptosis and DNA damages via oxidative stresses, such as intracellular ROS and MitoSOX/MitoMP [Manoalide, an antibiotic sesterterpenoid isolated from the marine sponge X/MitoMP ;–λ-carrageenan, a family of linear sulfated polysaccharides, proven to enhance the effect of radiotherapy by suppressing the survival and invasiveness of different cancer cell lines in vitro and in vivo through the Rac GTPase-activating protein 1 pathway ;–Ethanol, which was found to trigger a pro-survival autophagic response following the induction of oxidative and endoplasmic reticulum stress in colon cancer cells, and the activation of Nrf2 and HO-1 also leading to the acquisition of a more aggressive phenotype ;–Colchicum autumnale, whose enhanced anticancer effects and reduced cytotoxicity on colon cancer cells when delivered in the nanoformulated form was reported by AbouAitah et al. [Colchicine, an alkaloid present in the medicinal plant h et al. .The other articles and reviews addressed further cancer-related issues relevant to types of compounds of a different nature, specifically:In the Review section, Del Cornò et al.  discusseThe number of manuscripts published in this Special Issue indicates an active interest in research about the molecular/pharmacological mechanisms used by natural products exerting anti-tumoral effects which deserve further and deeper studies. I wish to thank all the contributors of this issue for sharing with us their experimental or reviewed data which will surely attract readers’ attention and encourage the publication of other high-quality papers in this field.


This Special Issue containing seminal contributions from international experts highlights the current understanding of Rho GTPases in cancer, with an emphasis on recognizing their central importance as critical targets for cancer therapy and for chemosensitization of current therapeutic strategies. A comprehensive review by Jung et al.  gives anNew regulatory mechanisms for Rho GTPases are presented in a review by Humphries et al. , who desNovel directions for Rho GTPase guanine nucleotide exchange factors (GEFs) are also highlighted in a number of peer-reviewed research articles. Using in silico analyses and in vitro experimental studies in keratinocytes, Lorenzo-Martin et al.  implicatTwo articles by Dr. Hendrick Ungefroren’s group contribute to the role of the constitutively active Rac1B splice variant in transforming growth factor (TGFβ) signaling ,9. The aFinally, Dyberg et al. implicate the downstream effector of Rho, Rho kinase (ROCK), in EMT and medulloblastoma growth by demonstrating that ROCK mRNA is preferentially expressed in metastatic tumors . They usIn summary, herein, you will find timely articles on the ubiquitous role of Rho GTPases in cancer and how understanding their mechanisms of action can lead to the design and development of targeted therapeutic strategies.


The worldwide increase of human life expectancy (“lifespan”) along with the concomitant rapid population aging represents the major social phenomena of the last century. This has substantially impacted our societies by virtue of the huge economic implications and public health challenges  definition of reference values; (2) identification of reference standards specific for the disease of interest ; (3) proper inclusion and contextualization within the diagnostic process; and (4) statistical processes supporting the whole framework. They concluded that various methodological issues remain to be addressed in order to perform an adequate and complete clinical validation of candidate CSF biomarkers for AD.Using cerebrospinal fluid (CSF) biomarkers for AD as a paradigmatic example, D'Anca et al. explored the current understanding of role of exosomes in physiological aging and age-related neurodegenerative diseases such as AD, Parkinson's Disease (PD) and frontotemporal dementia. Insights from the study of exosomes and their genetic cargo  have elevated their role beyond mere waste disposal function to fundamental mediators of intercellular communication, akin to the proverbial double-edge sword serving as Trojan horses of neurodegeneration vis-à-vis providing neuroprotection from neurodegeneration. Exosomes can be detected in many biological fluids, enhancing their appeal as potential sources of biomarkers suitable for use in clinical practice.Five papers in this Research Topic shed precious insights into emerging biomarkers from readily available biological samples. In their comprehensive review, Tay et al. prospectively studied serum levels of Dickkopf-1 (Dkk-1) in older adults with mild cognitive impairment (MCI) and mild-to-moderate AD. The findings revealed that Dkk-1 increased significantly from baseline amongst progressors, while non-progressors exhibited decremental Dkk-1 at 1 year, alluding to the putative role in MCI and AD progression of dysfunctional Wnt signaling through Dkk-1 antagonism. Sun et al. demonstrated that cofilin 2 expression was significantly increased in AD patients and different AD models , with good discriminatory ability that distinguish AD from healthy subjects and in differential diagnosis of AD from vascular dementia. By studying 391 cognitively normal subjects aged 23–91 years from Asia, USA and Europe, Lue et al. characterized the relationship between age and three plasma AD core biomarkers , provided the normal ranges of Aß species and t-Tau in plasma, and explicated the development of a dynamic relationship between these biomarkers from middle to old age. Lastly, Falconi et al. investigated the transcriptional regulation of the Adenosine A2A receptors (A2ARs) gene in human peripheral blood mononuclear cells obtained from PD patients and in the striatum of the 6-hydroxydopamine-induced PD mouse model. They reported an increase in A2AR mRNA expression and protein levels in both human cells and mice that is accompanied by histone acetylation and DNA methylation, paving the way for therapeutic interventions in future.Similarly, Li et al. shed light on the neural mechanisms of working memory deficits in MCI. By combining static and temporal dynamic examination of amplitude of low-frequency fluctuations (ALFF) from functional magnetic resonance imaging, they reported background network changes especially in the parietal and temporal lobes during working memory states in MCI. Similarly, Pur et al. enhanced understanding by demonstrating the moderating effect of cortical thickness on blood oxygen level-dependent (BOLD) signal variability age-related changes and highlight the importance of considering these effects when evaluating BOLDSD alternations across the lifespan. Kobayashi et al. examined whether dementia with Lewy Bodies (DLB) follows an AD-type trajectory whereby amyloid-ß deposition begins considerably before onset of dementia. They observed a low amyloid load in REM sleep behavioral disorder, a prodromal symptom of DLB, suggesting that this phenomenon does not always precede the onset of cognitive decline in DLB. Lastly, through the use of MRI voxel studies to examine neural substrates, Wu, Geng et al. implicated the left-precentral cortex and left inferior frontal gyrus areas that accounted for olfactory impairment in a cohort of AD and MCI patients in the Chinese Han population.Another prominent theme in this Research Topic was the contribution of advanced neuroimaging biomarkers. For instance, in terms of explicating mechanisms that underpin underlying pathophysiological processes, Wang, Hatashita et al. examined [18F]-flutemetamol (FMM), a fluorinated derivative of the prototypic [11C]-Pittsburgh Compound B (PIB), and demonstrated that [18F]-FMM PET imaging can track longitudinal changes in Aβ deposition across the AD spectrum, similarly to [11C]-PIB PET. Notably, they reported that the increase in Aβ deposition is not constant across the AD spectrum but faster in the predementia stage . Using dynamic [18F] florbetapir PET, Verfaillie et al. investigated the possibility that self-perceived cognitive decline (SCD), generally associated with a three- to six-fold increased risk of AD, may reflect an early symptom of Aβ related pathology. They concluded that Aβ load was associated with SCD related worries rather than subjective cognitive functioning per se .Other papers in this Research Topic illuminated the knowledge gap in our understanding about the role of misfolded protein accumulation in neurodegenerative diseases. The extracellular accumulation of amyloid beta (Aβ) peptide is the paradigmatic example, with recent neuroimaging capabilities in tracking Aβ deposition a fascinating way to distinguish if the observed Aβ accumulation is characteristic of the aging process or conversely, an etio-pathogenetic mechanism of AD . Furthermore, the study by Wu, Xu et al. provides proof-of-concept evidence that a topological examination of the structural connectivity networks with different parcellation schemes can provide important complementary AD-related information and thus contribute to a more accurate and earlier diagnosis of AD. Another promising technique involved amide proton transfer (APT) imaging as an imaging modality to detect tissue protein. In fact, Wang, Chen et al. adopting a modified APT method  in animal models, demonstrated that APT imaging could potentially provide molecular biomarkers for non-invasive diagnosis of AD. Using direct non-invasive brain network electrophysiological imaging, Zifman et al. established that this new technique can be used both to monitor brain aging and for early detection of abnormal changes leading to neurodegeneration.Other papers highlight advances in neuroimaging techniques that pave the way for fresh perspectives in early diagnosis of AD. Utilizing sophisticated measurements of cortical thickness with 3.0-Tesla MRI in a large population of cognitively normal individuals and patients with AD continuum, Lorio et al. described how the combination of these imaging modalities can be used as biomarkers of PD severity and prognosis which can be potentially useful for clinical trials. Likewise, Pelizzari et al. proposed the concomitant use of DTI to detect brain tissue microstructural alterations, together with arterial spin labeling (ASL) MRI to analyse abnormal cerebral perfusion patterns in PD. The study showed that DTI is a more sensitive technique than ASL to detect alterations in the basal ganglia in the early phase of PD, suggesting that a relationship between microstructural integrity and perfusion changes in the caudate may be present .Bioimaging techniques are also useful for the differential diagnosis of PD. Dopamine-transporter SPECT (DAT-SPECT), diffusion tensor imaging (DTI), and structural magnetic resonance imaging (sMRI) provide unique information about neurotransmitters and microstructural properties in PD. The longitudinal study of Chen et al. developed an innovative predictive model to evaluate white matter burden in hypertensive patients using metrics in 24-h ambulatory BP monitoring (systolic blood pressure [SBP] and daytime SBP standard deviation) and carotid ultrasound IMT. Leveraging upon the diagnostic potential of diabetic retinopathy (DR) afforded by its time-course which precedes the occurrence of T2DM cognitive impairment, Lu et al. verified the correlation between DR from fundus examination and T2DM cognitive impairment. They further established using magnetic resonance spectroscopy (1H-MRS) that this may be attributed to bilateral changes in hippocampal brain metabolism, alluding to the potential role of 1H-MRS for early diagnosis of T2DM cognitive impairment .Papers in this Research Topic also addressed the issue of age-associated co-morbidities, particularly cardiovascular diseases, and related risk factors such as Type II diabetes mellitus (T2DM), which often confound the measurement and interpretation of neurodegenerative biomarkers. Exploring the association between white matter hyperintensities with higher Intima-media thickness (IMT) and blood pressure variability, Bandera et al. suggested an omics approach  to accelerate the discovery of reliable biomarkers of HIV-associated neurocognitive disorders in the current era of virological suppression with modern anti-retroviral therapy. Similarly, persistent inflammation has been implicated as a cardinal mechanism of neurocognitive impairment and neurodegenerative features in Multiple Sclerosis (MS). Jakimovski et al. reported the novel finding that almost half the elderly subjects with MS are impaired on tests of cognitive processing speed or verbal fluency. Since the deficit in verbal fluency is not a typical hallmark of the cognitive profile associated with MS, it may represent a unique trait of old persons with MS neurocognitive profile .A major pathogenetic influence of co-morbidities or their treatment that have emerged in recent years is that of concomitant inflammatory processes that may alter disease manifestation and confound biomarker interpretation. An example is the cognitive decline observed in some HIV subjects during ART-mediated viral suppression, a phenomena which has been ascribed to cytokine-mediated inflammation. Barha et al. underlined the role of biological sex as a potential moderator in the relationship between physical activity and brain health in older adults. They suggested that different neurobiological mechanisms  as well as physiological adaptations to physical activities could be responsible for the differences in trajectories of decline observed in men and women . This highlights the importance of taking into consideration the potent moderating influence of sex differences for both cognitive and neural outcomes in future therapeutic and rehabilitative interventions involving physical activity in older adults.Lastly, there is growing interest in the role of physical activity and exercise as “medicine” that can be prescribed for brain health. In their comprehensive review, As guest editors for this Research Topic, we commend this collection of 23 articles to our readers as a timely addition and important contribution to the field. We are confident that this will spur further discourse and open avenues for further research into the rapidly evolving and fascinating area of biomarkers to disentangle physiological from pathological brain aging.FG, WL, and BA conceived the manuscript. FG and WL drafted the paper. BA critically appraised and edited the manuscript. All authors read and approved the final version of the paper.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Background: Growing evidence points out that a disturbance of gut microbiota may also disturb the gut–brain communication. However, it is not clear to what extent the alteration of microbiota composition can modulate brain function, affecting host behaviors. Here, we investigated the effects of gut microbiota depletion on emotional behaviors.Methods: Mice in the experimental group were orally administered ceftriaxone sodium solution  for 11 weeks. The open-field test and tail-suspension test were employed for the neurobehavioral assessment of the mice. Fecal samples were collected for 16s rDNA sequencing. The serum levels of cytokines and corticosterone were quantified using enzyme-linked immunosorbent assays. The immunohistochemistry method was used for the detection of brain-derived neurotrophic factor (BDNF) and c-Fos protein.Results: The gut microbiota for antibiotic-treated mice showed lower richness and diversity compared with normal controls. This effect was accompanied by increased anxiety-like, depression-like, and aggressive behaviors. We found these changes to be possibly associated with a dysregulation of the immune system, abnormal activity of the hypothalamic-pituitary-adrenal axis, and an alteration of neurochemistry.Conclusions: The findings demonstrate the indispensable role of microbiota in the gut–brain communication and suggest that the absence of conventional gut microbiota could affect the nervous system, influencing brain function. Gut microbiota, known as a reservoir of bacteria, not only plays an essential role in host digestion and energy metabolism but shapes host immunity  and given either sterile saline solution  or ceftriaxone sodium solution   intragastrically once a day for 11 consecutive weeks  were maintained (ten mice per cage) under a specific-pathogen-free (SPF) condition at 22–26°C, 40–60% humidity, and 12-h light-dark cycle. The mice were given 1 week to acclimate. All mice were fed with adequate food and clean water. At the end of adaptive phase, all mice  were randomly divided into two groups : open-field test (OFT) → tail-suspension test (TST). The OFT, which involves a low stress level, preceded the TST, which involves a high stress level  was used to extract fecal DNA. Then, the extraction was eluted using elution buffer and stored at −80°C until PCR amplification detection by LC-Bio . The V3-V4 region of the prokaryotic 16S rRNA gene was amplified with primers 338F (5′-ACTCCTACGGGAGGCAGCAG-3′) and 806R (5′-GGACTACHVGGGTWTCTAAT-3′)  axis in rodents. Rising corticosterone levels suggest increases in HPA axis activity  in the hippocampus and c-Fos in the amygdala. The whole process consisted of brain collection, sectioning, and immunolabeling. The details referred to previous literatures . The staining extent was divided into five grades according to the percentage of positive cells in the region: negative, 0–25, 26–50, 51–75, and 76–100%   regions of the hippocampus . Similarly, semi-quantification of c-Fos was performed by calculating the intensity score and fraction score in the CeC, CeL, and CeM regions of the amygdala. The immunohistochemical analysis was performed blind.P-values less than 0.05 were considered statistically significant.Data were expressed as the mean ± standard deviation or median (IQR) and analyzed by one-way ANOVA or Wilcoxon rank sum test in SPSS 22.0 software . p < 0.05)  .OFT is usually performed to assess locomotor activity and exploratory behavior  after gavage for 11 weeks  in the AB group   . In addiA slight decrease of BDNF in the CA1, CA3, and DG regions of the hippocampus was observed in the AB group compared to the CT group , Table 3The gut, a vulnerable but vital organ, is affected by different factors easily. Antibiotics are one of the common causes leading to gut disturbance, especially given the broad spectrum of antibiotics. Consistently, little is known about adverse effects of these antibiotics on health except for drug resistance. But, recently, medications with antibiotic have been reported to enhance the risk of allergies, inflammatory bowel diseases, obesity, and even mental diseases (Harris and Baffy, Ceftriaxone administration caused significant weight loss in the study. This is consistent with the previous finding that the weight gain of mice was delayed significantly following the ceftriaxone treatment (Miao et al., Proteobacteria became a dominant phylum, and the abundance of Bacteroidetes, Firmicutes, Actinobacteria, and Deferribacteres decreased. This result is supported by studies that ceftriaxone could characteristically decrease the alpha-diversity of the fecal microbiota accompanied with more Proteobacteria and less Bacteroidetes (Cheng et al., Proteobacteria is perceived as a diagnostic characteristic since it is closely related to colon epithelial oxygenation as well as the disruption of the gut anaerobic environment (Zhu et al., Firmicutes has become a controversial strain as some studies identified an increase in Firmicutes after ceftriaxone treatment, but others have demonstrated a declined Firmicutes in the ceftriaxone group (Cheng et al., Firmicutes (Huang et al., Firmicutes led to a reduction in short-chain fatty acids, which are an important physiological basis for low-level inflammation during depression (Huang et al., Bacteroidetes, as an important microbe for short-chain fatty acids, almost disappeared from the feces of the mice during exposure to ceftriaxone (Miao et al., Porphyromonadaceae, Escherichia, and Parabacteroides dominated the gut microbiota of the AB group mice, while Lactobacillus, Clostridiales, Acetatifactor, Bacteroidetes, Barnesiella, Helicobacter, Prevotella, Bacteroidales, and Alistipes were lowered. In line with this, some researchers have proposed that decreased Barnesiella after ceftriaxone gavage is a common and sensitive gut microbiota of the BALB/c mice and can be used as an indicator for assessing the balance of the gut microbiota (Zhao et al., Bacteroidetes is closely associated with digestion and interacts with the host's immune system, affecting the growth of other bacteria (Karlsson et al., Escherichia prevalence after oral antibiotic treatment has been reported for vancomycin and imipenem (Stokes, Escherichia could be beneficial or harmful as Escherichia is both a commensal and pathogenic inhabitant of a host's gastrointestinal tract. But most of the time, Escherichia is considered a potential pro-inflammatory bacteria (Liu et al., Porphyromonadaceae associates with mental deficits and cognitive disorders as well as anxiety-like behaviors in mice (Scott et al., Lactobacillus is known as a protective species against long-lasting metabolic disturbances and prevents gut dysbiosis, but was suppressed by ceftriaxone (Robles-Vera et al., Parabacteroide relates to the etiology of depression (Cheung et al., Ceftriaxone could result in a significant gut microbiota dysbiosis by killing most of the normal flora and providing the living space for other potential pathogens (Cheng et al.,  Stokes, , amoxici Stokes, , and met Stokes, . It is dBifidobacterium and Lactobacillus (Logan and Katzman, Lactobacillus fermentum strain NS9 administration not only normalized the composition of gut microbiota but reduced the anxiety-like behavior induced by ampicillin (Wang et al., Bifidobacterium infantis has also been identified in the rat separation model of depression (Desbonnet et al., Alterations in gut microbiota were accompanied by behavioral changes in the mice, including anxiety-like, depression-like, and aggressive behaviors. These behavioral changes cannot necessarily be a result of the direct toxic effect of ceftriaxone on the brain, since ceftriaxone is a non-absorbable antibiotic and usually given by injection. Previous studies have demonstrated the complex interaction between gut microbiota and the CNS; this is what is known as the MGB axis (Wang and Wang, Schistosoma mansoni showed a reduction in behaviors such as exploration and grooming (Sulaiman et al., Immune dysregulation was demonstrated by high levels of serum cytokines. This is supported by the evidence that inflammatory factors associate with a profile of behavioral changes (Capuron and Miller, Bifidobacterium species, have been demonstrated to be efficient in restoring HPA axis function (Moya-Pérez et al., Elevated corticosterone, one marker of HPA axis activation, was observed in the mice of the AB group (Borrow et al., Bifidobaterium adolescentis shows a promising anxiolytic and antidepressant property as it up-regulated BDNF expression by restoring the balance of gut microbiota (Guo Y. et al., Campylobacter jejuni (Goehler et al., The BDNF level showed a decreasing trend in the hippocampus of the AB group. According to the previous study, BDNF can maintain and promote development, differentiation and regeneration of neurons as well as affect learning and memory (Bercik et al., In general, we found that mice exposed to 11 weeks of ceftriaxone sodium treatment had a lower diversity and abundance of gut microbiota and showed more behavioral changes as compared to mice that were given normal saline. Dysregulation of the nerve-endocrine-immunological network may be a potential mechanism underlying abnormal behaviors induced by impaired gut microbiota. The study revealed the unknown side effects of antibiotics to a certain extent. Follow-up studies rebalancing the gut dysbacteriosis are required to further confirm the relationship between gut microbiota and brain function.PRJNA592623.The datasets generated for this study can be found in the NCBI, SRA accession: The animal study was reviewed and approved by Animal Care Advisory Committee of Sichuan University.ZZ and HY put forward the hypothesis, and BW, CT, LZ, and ML guided and supervised the experimental investigation and practices. LM, HW, and JL were responsible for data collection and analysis. YY provided pathologic diagnosis. ZZ was the main executer and involved in the entire research process, from proposal, planning and execution to implementation and composition. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Alzheimer's disease (AD) is the most common form of dementia. Its prevalence will significantly increase in the coming decades, whereas no efficient treatment is currently available. AD has two main neuropathological lesions: amyloid plaques and neurofibrillary tangles (NFTs). Amyloid plaques are composed of amyloid ß (Aß) peptides cleaved from the Amyloid Precursor Protein (APP) . Although genetic factors are estimated to represent 60% of the risk to develop LOAD, they remained largely unknown for a long time except for APOE  accounts for <5% of AD cases and is well characterized by mutations in three genes (The endolysosomal proteins listed above play critical roles at various steps of dynamin-dependent endocytosis and further steps . EndocytDynamin-dependent endocytosis is regulated by the interplay of the interactions between PRD of dynamin and SH3 domain-containing proteins (Ferguson and De Camilli, in vitro (Talaga et al., post-mortem brain tissues of AD and other tauopathies (Nishikawa et al., In vitro phosphorylation of tau by GSK3ß, a kinase abnormally activated in AD brains (Leroy et al., Given that endocytic proteins such as SHIP1, SHIP2, and SYNJ1 are PI-5-phosphatases involved in PI metabolism (Ramos et al., Many of the endocytic machinery proteins implicated in AD risk possess SH2, SH3 domains, and/or PRD and are involved in actin dynamics as well as in regulation of PIs. Because the endocytic machinery needs fine-tuned regulation of PIs and endocytic protein-protein interactions, the endocytic pathway must be highly vulnerable. Dysregulation of even one of these endocytic proteins could lead to significant endocytic abnormalities. Hyperphosphorylation of tau may further accelerate endocytic dysregulation. Genetic risk factors and tau pathology might well have profound impacts on synaptic functions, endolysosomal/autophagic pathways, and APP processing via dysfunction of endocytosis, actin network, and PI metabolism . EndolysKA constructed the main concept of the manuscript by exchanging opinions with the other authors. All authors participated in writing the manuscript. All authors contributed to manuscript revision, read and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Ion channels are expressed throughout nervous system development. The type and diversity of conductances and gating mechanisms vary at different developmental stages and with the progressive maturational status of neural cells. The variety of ion channels allows for distinct signaling mechanisms in developing neural cells that in turn regulate the needed cellular processes taking place during each developmental period. These include neural cell proliferation and neuronal differentiation, which are crucial for developmental events ranging from the earliest steps of morphogenesis of the neural tube through the establishment of neuronal circuits. Here, we compile studies assessing the ontogeny of ionic currents in the developing nervous system. We then review work demonstrating a role for ion channels in neural tube formation, to underscore the necessity of the signaling downstream ion channels even at the earliest stages of neural development. We discuss the function of ion channels in neural cell proliferation and neuronal differentiation and conclude with how the regulation of all these morphogenetic and cellular processes by electrical activity enables the appropriate development of the nervous system and the establishment of functional circuits adapted to respond to a changing environment. Nervous system development is a complex process in which neural cells undergo a transformation from neural stem cells to highly specialized neurons and glia to form different brain structures and spinal cord and establish circuitry that facilitates simple to advanced neural functions.Many cues have been recognized as drivers of the first steps in nervous system development. Morphogenetic proteins and growth factors regulate the number and type of neural cells as well as the morphogenesis of the neural tube. These include Sonic hedgehog (Shh), Bone Morphogenetic Proteins (BMPs), Wnts and trophic factors such as EGF, IGF, NGF, BDNF to mention few. Most of these factors are not exclusive to the organogenesis of the brain and spinal cord but instead support growth and act as morphogens of every tissue and organ in the developing embryo. Mechanistically, these developmental cues trigger a transcriptional combinatorial code that accompanies cells through their cell cycle progression and exit, differentiation and maturation  and IP3 receptor-operated Ca2+ release from stores in ventral domains of the spinal cord  and CaNeural activity is a feature of the maturing and mature nervous system, which during development facilitates the refinement of neural connections. The expression of ion channels in mature neurons is intrinsic to neuronal function. Diverse ion conductances are indispensable for neurotransmission, thus, the roles of different ion channels in synaptic function and neuronal excitability have been extensively studied. In contrast, the neurophysiological features of neural cells before synapse formation and before neuronal differentiation has not been as strong a focus of attention as those of mature neurons. Nevertheless, studies have argued that other forms of neural activity are present in neural cells throughout nervous system development Spitzer, .This activity may not be structured under a classical chemical synapse, but it is certainly dependent on ion channels gated by diverse mechanisms. Expression of voltage- and neurotransmitter-gated ion channels as well as transient receptor potential (TRP) channels, among others, is apparent in neural stem cells as early as neural plate stages (Abdul-Wajid et al., Here, we review studies addressing the pattern of expression of ion channels during development in neural cells before and during synapse formation. We compile investigations demonstrating a role for ion channels in neural cell proliferation, neural tube formation, and neuronal differentiation and discuss the consequences of having neural activity functioning in the early stages of nervous system development.Xenopus laevis spinal cord neurons are first recorded 8 h after exiting the cell cycle, when, these events manifest spontaneously, are Ca2+-dependent and long in duration (Spitzer and Lamborghini, + channel contributes to shorten the action potential duration and shifts it from Ca2+- to Na+-mediated (Barish, 2+, Na+ and K+ voltage-gated channel subunits for which their expression is developmentally regulated have been investigated (Harris, + current as development advances (Gurantz et al., 2+ currents are dominant at the earliest embryonic stage of chick limb motor neuron development, while later T currents decrease and N and L Ca2+ currents increase (McCobb et al., + and K+ currents in these motor neurons during embryonic development result in changes in action potential amplitude and duration, respectively, which in turn, modify the instructions of motor neurons to the muscle (McCobb et al., The excitable nature of neurons and muscle cells is dependent on the specific expression of ion channels and their subcellular localization in these cells. Seminal studies have investigated the developmental appearance of excitability in neurons and muscle cells through the progressive and differential expression of ion channels. Embryonic spinal cord neurons have served as a powerful model for the study of the ontogeny of excitability during development. Action potentials in  Barish, . The ide Harris, . In partXenopus spinal cord neurons and their activation contributes to the spontaneous Ca2+ spike activity in these cells before and during synapse formation (Root et al., In addition to voltage-gated ion channels and their fundamental role in contributing to the excitability of developing neurons and muscle cells, other types of ion channels are also present at the early stages of embryonic development. These channels are gated by diverse mechanisms, including notably, neurotransmitter-operated channels. GABA and glutamate receptors are expressed in immature Xenopus embryo (Spencer et al., Our recently published study shows that the cold-sensitive channel TRPM8 is expressed in the developing Xenopus neuroectodermal cells exhibit Ca2+ transients (Abdul-Wajid et al., 2+ channels (Abdul-Wajid et al., Expression of these diverse types of channels appears to start at even earlier stages of neural development, before the neural tube is formed. At neural plate stages, Transcripts and proteins for glutamate (Root et al., The process of neural tube formation consists of transforming a flat layer of cells known as the neural plate into a tubular structure from which the brain and spinal cord originate. The cellular events encompassing neural tube morphogenesis, all of which are tightly regulated, include neural plate cell proliferation, apicobasal polarization, apical constriction, elongation, cell intercalation, migration and differentiation (Wallingford et al., Xenopus laevis embryos. We demonstrated that during neural tube formation neural plate cells exhibit Ca2+ transients partly mediated by NMDA receptors. Inhibiting glutamate signaling, through pharmacological inhibition of NMDA receptors or downregulation of the GluN1 subunit, induces NTDs (2+ dynamics in the neural plate to a similar extent as inhibition of NMDA receptors. Moreover, preincubating embryos with NMDA partially rescues both the number of Ca2+ transients in the folding neural plate and the valproic acid-induced NTD phenotype (Sequerra et al., Our recently published study (Sequerra et al., ces NTDs . Valproices NTDs , also in2B interferes with mouse neural tube closure and morphogenesis (Choi et al., Many other neurotransmitter signaling systems have been identified as participants in the process of neural tube formation. Inhibiting serotonin receptors 5HTA and GABAB receptor ligands to pregnant rats alters embryos’ neural tube formation leading to NTDs (Briner, A receptors. Another chloride conductance-mediated neurotransmitter system that affects neural tube formation is glycine. Blocking glycinergic signaling by administering strychnine to pregnant rats results in embryos with anencephaly, anterior NTD (García-Alcocer et al., Administering GABA Briner, . The fac Briner, , which mSimilarly, enhancing or inhibiting NO levels by enhancing BMP signaling or inhibiting NO synthase in chicken embryos induces NTDs (Traister et al., Noradrenaline promotes neuronal differentiation by upregulating expression of N-tubulin in noggin-expressing neural plate cells, which is prevented by inhibiting α-adrenergic receptors (Messenger et al., 2+ signaling is a plausible common denominator for the action of diverse neurotransmitter systems on neural tube formation. Sources of Ca2+ can be intracellular from stores or extracellular through Ca2+ influx. Early studies in cultured rat embryos during cephalic neural fold elevation and neural tube closure assessed the role of Ca2+ influx and found that reducing it causes opening of the elevated neural folds (Smedley and Stanisstreet, 2+ transients in the neural plate, suggesting that additional mechanisms other than these ionotropic glutamate receptors contribute to neural plate cell Ca2+ dynamics during folding (Sequerra et al., Xenopus laevis embryos consisting of a Na+-dependent inward current which is stronger in the mid-lateral neural plate and decreases near the midline of the neural groove (Robinson and Stump, 2+ currents with characteristic kinetic parameters to transduce the signaling required for neural tube morphogenesis. Indeed, T-type Ca2+ channels are involved in neural tube formation and loss of them impairs neural fold closure in Xenopus laevis and Ciona embryos. Ca2+ influx through these channels is necessary for the regulation of cell adhesion during neural tube formation by ephrin signaling (Abdul-Wajid et al., 2+ transients seem to regulate apical actin dynamics in superficial neural plate cells of Xenopus laevis embryos, which, in turn, regulates neural plate cell apical constriction necessary for neural tube formation (Christodoulou and Skourides, Ca2+ signaling and downstream effectors recruited for neural plate folding and neural tube formation. The elucidation of these mechanisms will contribute to the delineation of safe therapies for the treatment of epilepsy during pregnancy.Further investigation is needed to identify the molecular mechanisms eliciting CaThe generation of the appropriate number of neurons and glial cells is essential not only during nervous system development but also in the adult brain where neurogenesis occurs in the hippocampus and olfactory bulb, and the peripheral nervous system during regeneration and remodeling. Thus, this is a highly regulated process because the dysregulated proliferation of neural stem cells can lead from tumors to neurodevelopmental disorders and birth defects like NTDs.2+ channels have all been implicated in regulating neural plate cell proliferation.The expression of ion channels during the early stages of development supports a role for them in the relevant cellular processes pertinent to these stages including neural cell proliferation. Different types of ion channels including voltage-gated, neurotransmitter-gated, TRPC and store-operated CaXenopus embryos, and, likely as a consequence, impairs lateromedial migration leading to NTDs. An increase in neural plate cell proliferation is also apparent by incubating embryos with the AED valproic acid (Sequerra et al., The action of glutamate-mediated regulation of neural plate cell proliferation is apparent as early as neural plate stages. We found that blocking NMDA receptor signaling increases neural plate cell proliferation in 2+ channels (LoTurco et al., In vitro studies on embryonic rat hippocampal neural progenitors reveal that stimulating glutamate signaling enhances neural cell proliferation or neuronal differentiation depending on the temporal pattern of NMDA receptor activation (Joo et al., Regulation of neural plate cell proliferation by glutamate-gated ion channels is present at later developmental stages during corticogenesis in the rodent brain. The effects of glutamate signaling on neural progenitor proliferation vary depending on the nervous system structure, the developmental stage and the type of model system and manipulation used in specific studies. Glutamate decreases the number of proliferating embryonic rat cortical cells through an AMPA/Kainate receptor-dependent mechanism that leads to depolarization of neural progenitors in the ventricular zone and activation of voltage-gated CaA receptor. In vivo studies in the neonatal mouse subventricular zone (Young et al., in vitro studies in cerebellar granule cells (Fiszman et al., A receptor-induced depolarization enhances neural progenitor cell proliferation. In contrast, activation of this receptor in the ventricular zone of the rat embryonic neocortex (LoTurco et al., 2+ channels and inhibits cell cycle progression. Even in the adult mouse brain, the GABAA receptor seems crucial in regulating neurogenesis by controlling hippocampal neural progenitors. GABAA receptor γ2 subunit-mediated signaling is responsible for controlling the experience-dependent transition between quiescence vs. proliferative states of the mouse hippocampal neural stem cell niche (Song et al., A receptor favors the quiescence of adult neural stem cells (Song et al., A receptor signaling (Dumitru et al., Another important neurotransmitter-gated ion channel that participates in neural progenitor cell proliferation is the GABAvia membrane depolarization. In particular, voltage-gated Ca2+ channels are pivotal for the regulation of neural progenitor proliferation and mouse embryonic cortical layer formation (Malmersjö et al., 2+ channels enable Ca2+ transients that propagate through a network of neural progenitors connected by gap junctions. Inhibiting this electrotonic transmission decreases neural progenitor proliferation suggesting that correlated Ca2+ transients are necessary for the regulation of neural progenitor proliferation (Malmersjö et al., 2+ channels in embryonic neural cells increases neural cell proliferation that has detrimental consequences to brain morphogenesis (Pai et al., 2+ channel-mediated signaling has, depends on developmental timing and location.Other ion channels directly involved in regulating neural cell proliferation include the voltage-gated ion channels. Moreover, some actions of the neurotransmitter receptor-gated ion channels seem to converge into the recruitment of voltage-gated ion channels 2+ channels, voltage-gated K+ and Na+ channels are involved in regulating neural cell proliferation. The voltage-gated Na+ channel β1 subunit is necessary for inhibiting granule cell precursor proliferation during the first week of mouse postnatal dentate gyrus development (Brackenbury et al., + channels was discovered recently in Drosophila larvae, where the single pore-forming voltage-gated Na+ channel α subunit, paralytic, regulates neural progenitor proliferation and survival (Piggott et al., + channel regulates neural stem cell proliferation in the subependymal zone and consequently neurogenesis in the mouse olfactory bulb (Petrik et al., + channel α-subunit Kv6.4 regulates the proliferation of cells lining the embryonic brain ventricles (Shen et al., v6.4 increases it. Moreover, the Kv6.4 action appears antagonized by the expression of a homolog to the delayed rectifier K+ channel subunit Kv2.1, for which gain and loss of function manipulations cause the opposite effects on neural progenitor proliferation and ventricular brain development (Shen et al., In addition to Ca2+-dependent mechanism (Fiorio Pla et al., 2+ release-activated Ca2+ channels that enable store-operated Ca2+ entry in these cells mediated by Orai1 and STIM1. Downregulating the expression of these molecules decreases in vitro and in vivo proliferation of neural stem cells (Somasundaram et al., Non-voltage-gated ion channels have also been implicated in regulating neural cell proliferation. For instance, TRPC1 participates in bFGF/FGFR1-mediated proliferation of embryonic rat neural stem cells through a Ca+ channels and membrane depolarization, oligodendrocyte progenitor proliferation is regulated by controlling the progression of the cell cycle at the G1 phase, likely through a cAMP and cyclin-dependent kinase inhibitors p27Kip1 and p21CIP1 mechanism (Ghiani et al., A receptor-triggered hyperpolarization in embryonic and neural crest stem cells, S-phase checkpoint kinases of the phosphatidylinositol-3-OH kinase-related kinase family and, subsequently the histone variant H2AX, regulate proliferation (Andäng et al., + channel by fluid flow elicits Ca2+ dynamics through Ca2+ release-activated channels that activate ERK, regulating neural cell proliferation in the mouse subependymal zone in the olfactory bulb (Petrik et al., in vivo activates ERK in neural plate cells during Xenopus neural tube formation. In turn, constitutive activation of ERK during neural plate folding completely rescues the NTD phenotype induced by dysregulated neural plate cell proliferation due to blocking NMDA receptors or incubation with the AED valproic acid (Sequerra et al., The specific effect that is triggered downstream of all these ion channels on cell proliferation varies among these studies. This is likely due to differences in downstream signaling, recruitment of molecular partners, environmental milieu and maturational status of the cells subjected to these ion channel-triggered signaling mechanisms. There are many downstream signaling effectors reported to mediate ion channel-dependent regulation of neural cell proliferation. For instance, downstream of voltage-gated K+ channel, which in turn assigns a functional neuronal identity to developing nervous system cells (Maue et al., 2+ spikes generated by voltage-gated Ca2+ and Na+ channels in embryonic neurons (Gu et al., Xenopus laevis development (Gu and Spitzer, 2+ spikes that embryonic neurons exhibit is important for the type of neurotransmitter spinal cord neurons express, following a homeostatic rule by which higher spike frequency leads to expression of inhibitory neurotransmitters while suppression of these spikes results in expression of excitatory neurotransmitters (Borodinsky et al., 2+ waves, dependent on both extracellular Ca2+ and Ca2+ release from ion channels present in intracellular Ca2+ stores such as ryanodine receptors, are apparent in growth cones of developing neurons (Gu et al., It has been long recognized that expression of ion channels and prominently voltage-gated ion channels, their density, clustering, and subcellular localization are at the core of what distinguishes a neuron from other cell types. This fundamental question was addressed by pioneering studies from the Mandel lab when they cloned the transcription factor REST and identified it as a silencer element active in nonneuronal cells. In contrast, the absence of REST in neurons allows for the expression of the type II Na+ channels have been at the center of the process of neuronal differentiation in a variety of central nervous system structures and species. Weaver mice carry a mutation in a G-protein coupled inward rectifier K+ channel, GIRK2, that affects the pore-forming domain of the protein and impairs cerebellar granule neuron differentiation immediately after cell cycle exit (Patil et al., + channels, in turn, is a key determinant of the spontaneous Ca2+ activity that developing neurons exhibit. For instance, K+ channels like the small conductance Ca2+-activated K+ channel SK2 are expressed in immature Purkinje cells in the postnatal rat cerebellum and establish a feedback loop with Ca2+ influx and activation of Ca2+ channels to regulate the spatiotemporal features of Ca2+ transients in developing neurons (Cingolani et al., + channel during a critical period of development (Ribera and Spitzer, K2+ activity that differentiating neurons exhibit during development either directly or indirectly. Another instance of direct regulation of Ca2+ activity in developing neurons that connects the process of differentiation with the environment in which embryos develop is represented by the role of TRPM8, cold-sensitive channel, in spinal cord neuron differentiation (Spencer et al., 2+ spike frequency in ventral spinal cord neurons of Xenopus laevis embryos. In turn, Ca2+ spike activity regulates the expression of Hb9 (Spencer et al., +, Ca2+, and K+ channels and the downstream interaction between Ca2+ and cAMP dynamics (Peng et al., While all these studies are focused on specific ion channels and their role in different aspects of neuronal differentiation, they converge on shaping the spontaneous Ca2+ dynamics downstream of neurotransmitter receptor activation. NMDA receptor function is necessary for the development of dendritic arbors in differentiating retinotectal neurons and to establish appropriate retinotectal topographic maps (Cline and Constantine-Paton, 2+ channels to regulate neuritogenesis (Maric et al., Neurotransmitter-gated channels that regulate neuronal differentiation also involve Ca2+ channel-dependent mechanism that results in a higher number of calbindin-expressing mouse hippocampal pyramidal neurons when Neurotrophin-3 signaling is enhanced and lower when it is decreased (Boukhaddaoui et al., 2+]i in growth cones of rat cerebellar granule cells and Xenopus spinal cord neurons through the recruitment of TRPC channel activity (Li et al., Neurotransmitter modulation of neuronal differentiation through direct or indirect regulation of ion channel activity is a function shared by neurotrophic factors. Neurotrophin 3 signaling regulates the specification of neuronal phenotype through a voltage-gated CaXenopus retinal ganglion cells that direct their growth towards softer tissue (Koser et al., Ion channels can be mechanically gated and participate in the differentiation of developing neurons. Piezo 1, a mechanosensitive channel, mediates axonal growth and pathfinding of + channels are involved in regulating axonal outgrowth and morphology. In zebrafish, knockdown of the Nav1.6a alters axonal outgrowth and morphology of dorsally and ventrally projecting secondary motor neurons (Pineda and Ribera, Voltage-gated Na2+ transients recruit activity-dependent transcription factors like CREB (Belgacem and Borodinsky, 2+ channels are recruited downstream of the clustering of the neural cell adhesion molecule 2, inducing submembrane Ca2+ spikes that in turn activate the protein tyrosine kinase c-Src, which results in CaMKII activation and increases in filopodia density, neurite outgrowth and branching (Sheng et al., Downstream of channel activity local and whole-cell effectors are recruited to change, for example, directionality and growth rate of neurites and neurotransmitter specification, respectively. Cav1.2 that translocates to the nucleus and acts as a transcription factor controlling rat neuronal differentiation (Gomez-Ospina et al., 2+ permeation and is dependent on localized RhoA activation (Krey et al., 2+-dependent imbalance in the numbers of subtypes of differentiated cortical neurons (Panagiotakos et al., Alternatively, ion channels may trigger downstream signaling relevant for neuronal differentiation independently of ion permeation. For example, in some instances, the channel itself regulates transcription, as shown for the C-terminal fragment of Ca2+ dynamics in developing neural cells. The participation of neural activity via ion channel expression throughout neural development poses the question of whether this makes the developing nervous system more vulnerable to “hijacking” of the necessary signaling mechanisms by exogenous unwanted factors. For instance, we have shown that incubating embryos with the AED valproic acid interferes with Ca2+ dynamics in neural plate cells and results in NTDs (Sequerra et al., 2+ activity-dependent mechanism operating in embryonic spinal cord neurons, exhibit an enhanced escape locomotor performance compared to siblings grown at warm temperatures when tested at cold temperatures (Spencer et al., The studies presented demonstrate that ion channels are expressed from the very first stages of neural development. Furthermore, the signaling mechanisms triggered by these ion channels participate in all the relevant cellular processes of early development including neural cell proliferation and neuronal differentiation, mostly through imprinting specific spatiotemporal CaRG, KS and LB wrote the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Dear Editor,Hi CD8+ peripheral T-cells for a cytolytic gene signature. Expression of cytotoxic genes in the peripheral blood was associated with worse prognosis, which may be explained by persistent micrometastatic disease that finally leads to recurrence . The aual., 2020). Numerous studies have analyzed the association of breast cancer with outcome, including the spontaneous course of disease and response to chemotherapy (Sparano et al., 2018; Wang etThe author declares no conflict of interest.