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Toll‐like receptor 2 (TLR2) belongs to the TLR protein family that plays an important role in the immune and inflammation response system. While TLR2 is predominantly expressed in immune cells, its expression has also been detected in the brain, specifically in microglia and astrocytes. Recent studies indicate that genomic deletion of TLR2 can result in impaired neurobehavioural function. It is currently not clear if the genomic deletion of TLR2 leads to any alterations in the microstructural features of the brain. In the current study, we noninvasively assess microstructural changes in the brain of TLR2‐deficient (tlr2−/−) zebrafish using state‐of‐the art magnetic resonance imaging (MRI) methods at ultrahigh magnetic field strength (17.6 T). A significant increase in cortical thickness and an overall trend towards increased brain volumes were observed in young tlr2−/− zebrafish. An elevated T2 relaxation time and significantly reduced apparent diffusion coefficient (ADC) unveil brain‐wide microstructural alterations, potentially indicative of cytotoxic oedema and astrogliosis in the tlr2−/− zebrafish. Multicomponent analysis of the ADC diffusivity signal by the phasor approach shows an increase in the slow ADC component associated with restricted diffusion. Diffusion tensor imaging and diffusion kurtosis imaging analysis revealed diminished diffusivity and enhanced kurtosis in various white matter tracks in tlr2−/− compared with control zebrafish, identifying the microstructural underpinnings associated with compromised white matter integrity and axonal degeneration. Taken together, our findings demonstrate that the genomic deletion of TLR2 results in severe alterations to the microstructural features of the zebrafish brain. This study also highlights the potential of ultrahigh field diffusion MRI techniques in discerning exceptionally fine microstructural details within the small zebrafish brain, offering potential for investigating microstructural changes in zebrafish models of various brain diseases. [ABSTRACT FROM AUTHOR]

Epimedium is thought to enhance the integrity of tendons and bones, ease joint discomfort and rigidity and enhance kidney function. Although glucocorticoids are commonly used in clinical practice, the mechanism by which the active compound Epimedin C (EC) alleviates glucocorticoid‐induced osteoporosis (GIOP) is not well understood. The therapeutic potential of EC in treating GIOP was evaluated using alizarin red S staining, calcein immersion and fluorescence imaging, and bone mineralization, bone mass accumulation and bone density in zebrafish larvae were determined. Using the Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, the key signalling pathways related to bone development were identified. A protein–protein interaction network (PPIN) was constructed to identify osteoclast characteristic genes and the findings were verified using real‐time quantitative PCR (RT‐qPCR). The bone tissue damage caused by prednisolone was reduced by EC. It also altered physiological processes, improved bone density, boosted mineralization and increased bone mass and activity. Subsequent empirical investigations showed that EC impacted the major signalling pathways involved in bone development, such as osteoclast differentiation, oestrogen, MAPK, insulin resistance, PPAR and AMPK signalling pathways. It also decreased the expression of genes typical of osteoclasts. The results of our study uncover a previously unknown function of EC in controlling bone formation and emphasize the potential of EC as a therapeutic target. The osteoprotective effect of EC indicates its potential as a cost‐effective strategy for treating GIOP. [ABSTRACT FROM AUTHOR]

Platinum‐based antineoplastic drugs, including cisplatin, carboplatin, and oxaliplatin, are widely used in the treatment of various cancers. Ototoxicity is a common adverse effect of platinum‐based drugs. Ototoxicity leads to irreversible hearing impairment. We hypothesize that different platinum‐based drugs exhibit varying ototoxic concentrations, time effects, and ototoxic mechanisms. We tested this hypothesis by using a zebrafish model (pvalb3b: TagGFP) to assess the viability of hair cells collected from zebrafish larvae. Cisplatin, carboplatin, and oxaliplatin were administered at dosages of 100, 200, or 400 μM, and the ototoxic effects of these drugs were assessed 1, 2, or 3 h after administration. Fm4‐64 and a TUNEL assay were used to label the membranes of living hair cells and to detect cell apoptosis, respectively. We observed that >50% of hair cells were damaged at 1 h after cisplatin (100 μM) exposure, and this ototoxic effect increased at higher dosages and over time. Owing to the smaller ototoxic effects of carboplatin and oxaliplatin, we conducted higher‐strength and longer‐duration experiments with these drugs. Neither carboplatin nor oxaliplatin was obviously ototoxic, even at 1600 μM and after 6 h. Moreover, only cisplatin damaged the membranes of the hair cells. Cell apoptosis and significantly increased antioxidant gene expression were observed in only the cisplatin group. In conclusion, cisplatin significantly damages sensory hair cells and has notable dosage and time effects. Carboplatin and oxaliplatin are less ototoxic than cisplatin, likely due to having different ototoxic mechanisms than cisplatin. [ABSTRACT FROM AUTHOR]

Missense mutations in MYOT encoding the sarcomeric Z‐disk protein myotilin cause three main myopathic phenotypes including proximal limb‐girdle muscular dystrophy, spheroid body myopathy, and late‐onset distal myopathy. We describe a family carrying a heterozygous MYOT deletion (Tyr4_His9del) that clinically was characterized by an early‐adult onset distal muscle weakness and pathologically by a myofibrillar myopathy (MFM). Molecular modeling of the full‐length myotilin protein revealed that the 4‐YERPKH‐9 amino acids are involved in local interactions within the N‐terminal portion of myotilin. Injection of in vitro synthetized mutated human MYOT RNA or of plasmid carrying its cDNA sequence in zebrafish embryos led to muscle defects characterized by sarcomeric disorganization of muscle fibers and widening of the I‐band, and severe motor impairments. We identify MYOT novel Tyr4_His9 deletion as the cause of an early‐onset MFM with a distal myopathy phenotype and provide data supporting the importance of the amino acid sequence for the structural role of myotilin in the sarcomeric organization of myofibers. [ABSTRACT FROM AUTHOR]

Giant axonal neuropathy (GAN) is a fatal neurodegenerative disorder for which there is currently no treatment. Affecting the nervous system, GAN starts in infancy with motor deficits that rapidly evolve toward total loss of ambulation. Using the gan zebrafish model that reproduces the loss of motility as seen in patients, we conducted the first pharmacological screening for the GAN pathology. Here, we established a multilevel pipeline to identify small molecules restoring both the physiological and the cellular deficits in GAN. We combined behavioral, in silico, and high‐content imaging analyses to refine our Hits to five drugs restoring locomotion, axonal outgrowth, and stabilizing neuromuscular junctions in the gan zebrafish. The postsynaptic nature of the drug's cellular targets provides direct evidence for the pivotal role the neuromuscular junction holds in the restoration of motility. Our results identify the first drug candidates that can now be integrated in a repositioning approach to fasten therapy for the GAN disease. Moreover, we anticipate both our methodological development and the identified hits to be of benefit to other neuromuscular diseases. Synopsis: To date, there is no treatment for giant axonal neuropathy (GAN), a fatal neurodegenerative disease leading to loss of ambulation in young patients. Due to the lack of effective rodent models, the gan zebrafish was used to conduct the first pharmacological screening for therapeutic repurposing.The gan zebrafish is the first robust model of the GAN disease, exhibiting the loss of motility found in patients.A multi‐level methodology was designed to integrate behavioral (motility), computational (drug‐target functions) and cellular (NMJ compartments) readouts.A novel high‐content automated imaging analysis was developed to quantify the beneficial effect of the Hits at the NMJ.This tailored screening pipeline identified five Hits restoring the motility of the gan zebrafish through the stabilization of the NMJ. [ABSTRACT FROM AUTHOR]

ZC4H2 (MIM# 300897) is a nuclear factor involved in various cellular processes including proliferation and differentiation of neural stem cells, ventral spinal patterning and osteogenic and myogenic processes. Pathogenic variants in ZC4H2 have been associated with Wieacker‐Wolff syndrome (MIM# 314580), an X‐linked neurodevelopmental disorder characterized by arthrogryposis, development delay, hypotonia, feeding difficulties, poor growth, skeletal abnormalities, and dysmorphic features. Zebrafish zc4h2 null mutants recapitulated the human phenotype, showed complete loss of vsx2 expression in brain, and exhibited abnormal swimming and balance problems. Here we report 7 new patients (four males and three females) with ZC4H2‐related disorder from six unrelated families. Four of the 6 ZC4H2 variants are novel: three missense variants, designated as c.142T>A (p.Tyr48Asn), c.558G>A (p.Met186Ile) and c.602C>T (p.Pro201Leu), and a nonsense variant, c.618C>A (p.Cys206*). Two variants were previously reported : a nonsense variant c.199C>T (p.Arg67*) and a splice site variant (c.225+5G>A). Five patients were on the severe spectrum of clinical findings, two of whom had early death. The male patient harboring the p.Met186Ile variant and the female patient that carries the p.Pro201Leu variant have a relatively mild phenotype. Of note, 4/7 patients had a tethered cord that required a surgical repair. We also demonstrate and discuss previously under‐recognized phenotypic features including sleep apnea, arrhythmia, hypoglycemia, and unexpected early death. To study the effect of the missense variants, we performed microinjection of human ZC4H2 wild‐type or variant mRNAs into zc4h2 null mutant zebrafish embryos. The p.Met186Ile mRNA variant was able to partially rescue vsx2 expression while p.Tyr48Asn and p.Pro201Leu mRNA variants were not. However, swimming and balance problems could not be rescued by any of these variants. These results suggest that the p.Met186Ile is a hypomorphic allele. Our work expands the genotypes and phenotypes associated with ZC4H2‐related disorder and demonstrates that the zebrafish system is a reliable method to determine the pathogenicity of ZC4H2 variants. [ABSTRACT FROM AUTHOR]

In vivo toxicological studies are currently necessary to analyze the probable dangers of quantum dots (QDs) to the environment and human safety, due to the fast expansion of QDs in a range of applications. Because of its high fecundity, cost‐effectiveness, well‐defined developmental phases, and optical transparency, zebrafish has long been considered the "gold standard" for biosafety assessment of chemical substances and pollutants. In this review, the advantages of using zebrafish in QD toxicity assessment were explored. Then, the target organ toxicities such as developmental toxicity, immunotoxicity, cardiovascular toxicity, neurotoxicity, and hepatotoxicity were summarized. The hazardous effects of different QDs, including cadmium‐containing QDs like CdTe, CdSe, and CdSe/ZnS, as well as cadmium‐free QDs like graphene QDs (GQDs), graphene oxide QDs (GOQDs), and others, were emphasized and described in detail, as well as the underlying mechanisms of QDs generating these effects. Furthermore, general physicochemical parameters determining QD‐induced toxicity in zebrafish were introduced, such as chemical composition and surface coating/modification. The limitations and special concerns of using zebrafish in QD toxicity studies were also mentioned. Finally, we predicted that the utilization of high‐throughput screening assays and omics, such as transcriptome sequencing, proteomics, and metabolomics will be popular topic in nanotoxicology. In this review, the advantages of using zebrafish in QD toxicity assessment were explored. Then, the target organ toxicities were summarized. The hazardous effects of different QDs, including cadmium‐containing QDs and cadmium‐free QDs, were emphasized and described in detail, as well as the underlying mechanisms of QDs generating these effects. Furthermore, general physicochemical parameters determining QD‐induced toxicity in zebrafish were introduced. The limitations and special concerns of using zebrafish in QD toxicity studies were also mentioned. [ABSTRACT FROM AUTHOR]

Humans are exposed to obesity causing Bisphenol A in various ways, especially through diet and food containers. Bioactive peptides are already reported to have antioxidant, antidiabetic, and antiobesity properties, which can mimic the role of mediators involved in obesity prevention. The protective effect of a short molecule or peptide, WL15 from cysteine and glycine‐rich protein 2 of a teleost of aquatic resource on Bisphenol A (BPA)‐induced lipid accumulation in zebrafish larvae was investigated. BPA exposure disrupted the antioxidant enzymes, apoptosis, and nitric oxide and led to changes in biochemical markers including alkaline phosphatase, lactate dehydrogenase, lipid peroxidation, glutathione S‐transferases, glutathione peroxidase, and reduced glutathione. However, WL15 inhibited the overproduction of oxidative stress, which correlates with its lipid‐lowering potential. BPA‐induced lipid accumulation in zebrafish showed an increase in triglyceride, cholesterol, and glucose level; simultaneously, WL15 treatment significantly reduced such accumulation in zebrafish. Evidenced by Oil red O staining and Nile red assay, WL15 inhibited lipid accumulation. At the same time, WL15 at 50 µM increases 2‐(N‐[7‐nitrobenz‐2‐oxa‐1,3‐diazol‐4‐yl]amino)‐2‐deoxy‐d‐glucose (2NBDG) glucose uptake in zebrafish. In addition, gene expression studies in zebrafish larvae demonstrated that the WL15 peptide could play a crucial role in preventing lipid accumulation by downregulating the expression of lipogenesis‐specific genes. These results revealed an interesting and novel property of WL15, suggesting its potential application in preventing lipid accumulation through the hypolipidemic and antioxidant properties. [ABSTRACT FROM AUTHOR]

Aims: TFG‐related axonal Charcot–Marie–Tooth (CMT) disease is a late‐onset, autosomal dominant, hereditary motor, and sensory neuropathy characterized by slowly progressive weakness and atrophy of the distal muscles. The objective of this study was to determine the common pathogenic mechanism of TFG‐related CMT type 2 (CMT2) caused by different mutations and establish a direct association between TFG haploinsufficiency and neurodegeneration. Methods: Three individuals carrying the TFG p.G269V mutation but with varying disease durations were studied. The effect of the p.G269V mutation was confirmed by analyzing protein samples extracted from the blood of two individuals. The functional consequences of both CMT2 mutant gene products were evaluated in vitro. The effect of TFG deficiency in the nervous system was examined using zebrafish models and cultured mouse neurons. Results: Overexpression of p.G269V TFG failed to enhance soluble TFG levels by generating insoluble TFG aggregates. TFG deficiency disrupted neurite outgrowth and induced neuronal apoptosis both in vivo and in vitro and further impaired locomotor capacity in zebrafish, which was consistent with the phenotype in patients. Wnt signaling was activated as a protective factor in response to TFG deficiency. Conclusion: CMT2‐related TFG mutation induces TFG haploinsufficiency within cells and drives disease by causing progressive neurite degeneration. [ABSTRACT FROM AUTHOR]

Background: Fruit acids have long been recognized as highly effective actives with world‐wide popularity, covering skin peeling, anti‐acne, anti‐wrinkle applications, and skin depigmentation. Aims: There are complicated interconnections between the fruit acid formula and skin pigmentation behaviors. However, the lack of systematic researches on multi‐ingredient formula restricted our understanding on its mechanism. Therefore, it is of great necessity to study the interactions and cascades among components, potential gene targets, signaling pathways, and biological processes via a system biology approach. Methods: We used system biology, molecular fingerprint, and structural biology to calculate and collect target information of the functional formula. Gene Ontology (GO) enrichment analysis was further applied to study the biological processes and pathways of the formula, and a multi‐level network model of "component ‐ molecular target ‐ signaling pathway ‐ skin disease" was established. Besides, the zebrafish model was utilized to verify the formula. Results: We obtained 69 hub targets by constructing a protein–protein interaction (PPI) network based on the intersection between multi‐fruit acids formula (mandelic acid, lactobionic acid, niacinamide, and hydroxytyrosol) and skin indications targets (whitening‐sebum balance). In vitro zebrafish models, including pigmentation, antioxidant, and radiation protection models, showed that the current formula significantly alleviated pigmentation intensity and intracellular free radical content, thus proving the efficacy of skin brightening and UV irradiation protection. Conclusions: This research uncovered the underlying mechanism of multi‐fruit acids formula and predicted its function in skin brightening and UV irradiation prevention. [ABSTRACT FROM AUTHOR]

Effects of casein on the stability, antioxidant activity, and bioavailability of lotus anthocyanins were investigated. Casein could inhibit the unsatisfactory pH‐induced color change of lotus anthocyanins, and improved their photo, oxidation, and thermal stabilities. During the simulated digestion, the anthocyanin retention increased from 65.39 to 76.14 mg C3G/L with the protection of casein, while the DPPH and ABTS scavenging activities of lotus anthocyanins with casein increased to 62.33% and 46.58%, respectively. However, casein with lower concentration showed a better protective effect on lotus anthocyanins due to its self‐aggregation tendency at high dose. The zebrafish model further verified that casein could enhance the bioavailability of lotus anthocyanins. Furthermore, molecular docking revealed that casein could interact with anthocyanin by hydrogen bond and hydrophobic interaction, which led to the stronger stability and bioavailability of lotus anthocyanins. The results conveyed that casein could be used as a wall material to protect anthocyanins. Practical applications: Anthocyanins are natural colorants with multiple biological activities, but the poor stability during processing and digestion limits their application in food industry. In the present research, casein exhibited conspicuous ability to enhance the stability of lotus anthocyanins toward detrimental conditions. Additionally, casein could preserve anthocyanins from degradation during digestion and thus improve the bioavailability. These findings indicated that casein could serve as a potential carrier for encapsulating and delivering anthocyanins. The better stability and bioavailability would promote the application of anthocyanins in food products and human health. [ABSTRACT FROM AUTHOR]

Human health may benefit from the study of natural compounds and phytoconstituents that can protect from inflammation. We investigated Nimbin (N1), a member of the ring C Seco‐tetranortriterpenoids family, and its semi‐natural analog deacetyl Nimbin namely N2 and N3 for their anti‐inflammatory properties. As key findings, N1, N2, and N3 were able to improve wound healing by cell proliferation in a period of 24 h and were able to reduce the reactive oxygen species (ROS) production in Madin–Darby Canine Kidney cells which were screened using dichloro‐dihydro fluorescein diacetate (DCF‐DA) staining. When the zebrafish larvae were subjected to DCF‐DA assay N1, N2, and N3 were able to substantially reduce the ROS levels in a dose‐dependent manner. In zebrafish larvae, the cell death indicates the fluorescent intensity due to acridine orange staining that was found to be dramatically decreasing upon the treatment of N1, N2, and N3. The cell membrane lipid peroxidation levels were also reduced in a dose‐dependent manner upon the treatment of Nimbin and its analogs indicating lesser blue fluorescent levels. Among the Nimbin and its analogs, N2 was subjected to have better activity. To confirm the activity of N1, N2, and N3, in silico characterization was performed using Density functional theory and molecular docking. As a result, N2 exhibited the lowest electronegative value and highest binding energy when docked with anti‐inflammatory and antioxidant proteins CAT, COX, GP, IL‐1, and MPO. Furthermore, the therapeutic potential of N2 must be explored at the molecular level as well as in clinical studies for the treatment of inflammation‐associated diseases. [ABSTRACT FROM AUTHOR]

Diphlorethohydroxycarmalol (DPHC) is a marine polyphenolic compound derived from brown alga Ishige okamurae. A previously study has suggested that DPHC possesses strong mushroom tyrosinase inhibitory activity. However, the anti‐melanogenesis effect of DPHC has not been reported at cellular level. The objective of the present study was to clarify the melanogenesis inhibitory effect of DPHC and its molecular mechanisms in murine melanoma cells (B16F10) and zebrafish model. DPHC significantly inhibited tyrosinase activity and melanin content dose‐dependently in α‐melanocyte stimulating hormone (α‐MSH)‐stimulated B16F10 cells. This polyphenolic compound also suppressed the expression of phosphorylation of cAMP response element‐binding protein (CREB) by attenuating phosphorylation of cAMP‐dependent protein kinase A, resulting in decreased MITF expression levels. Furthermore, DPHC downregulated MITF protein expression levels by promoting the phosphorylation of extracellular signal‐regulated kinase. It also inhibited tyrosinase, tyrosinase‐related protein 1 (TRP‐1), and TRP‐2 in α‐MSH stimulated B16F10 cells. In in vivo studies using zebrafish, DPHC also markedly inhibited melanin synthesis in a dose‐dependent manner. These results demonstrate that DPHC can effectively inhibit melanogenesis in melanoma cells in vitro and in zebrafish in vivo, suggesting that DPHC could be applied in fields of pharmaceutical and cosmeceuticals as a skin‐whitening agent. Significance of study: The present study showed for the first time that DPHC could inhibit a‐MSH‐stimulated melanogenesis via PKA/CREB and ERK pathway in melanoma cells. It also could inhibit pigmentation in vivo in a zebrafish model. This evidence suggests that DPHC has potential as a skin whitening agent. Taken together, DPHC could be considered as a novel anti‐melanogenic agent to be applied in cosmetic, food, and medical industry. [ABSTRACT FROM AUTHOR]

Developmental and epileptic encephalopathy (DEE) is a severe encephalopathy in infants and early childhood. In this study we reported a recurrent de novo variant (c.3985C>T, p.R1330W) in HECW2 (HECT, C2 and WW domain containing E3 ubiquitin protein ligase 2) (MIM# 617245) identified by screening 240 patients with DEE and summarized clinical features of published DEE patients with HECW2 variants. Functionally, transcriptional knockdown of zebrafish hecw2a led to early morphological abnormalities in the brain tissues. These results suggest a potential functional link between HECW2 dysfunction and brain development. [ABSTRACT FROM AUTHOR]

Metal and metal oxide nanoparticles have been widely used in catalytic, electronic and biomedical fields. It is necessary to investigate their toxicity and potential hazards to human and aquatic ecosystems. Zebrafish (Danio rerio), as a promising animal model, has been increasingly utilized to assess the toxicity of nanoparticles. Zebrafish has numerous characteristics for toxicity evaluation, such as short life cycle and high fecundity. This review describes the advantages of using zebrafish in the toxicity assessment of metal and metal oxide nanoparticles. Then we focus on the toxic effects, particularly the acute toxicity and the chronic ones, induced by nanoparticles in zebrafish. Target organ toxicities are also mentioned, including immunotoxicity, developmental toxicity, neurotoxicity, reproductive toxicity, cardiovascular toxicity and hepatotoxicity. The toxic effects of selected metal nanoparticles, including Au, Ag, Cu, and metal oxide nanoparticles such as TiO2, Al2O3, CuO, NiO and ZnO, as well as the underlying mechanisms of nanoparticles causing these effects, are also highlighted and described in detail. Furthermore, we introduce the general factors that affect nanoparticle‐induced toxicity in zebrafish. The drawbacks and advantages of using the zebrafish model in nanotoxicity studies are also argued. Finally, we suggest that the application of zebrafish to assess chronic toxicity of metal and metal oxide nanoparticles and the joint toxicity of metal and metal oxide nanoparticles and other pollutants could be hot topics in nanotoxicology. This review describes the advantages of using zebrafish (Danio rerio) in the toxicity assessment of metal and metal oxide nanoparticles. Focus is then on the acute toxicity and the chronic toxicity induced by nanoparticles in zebrafish. Target organ toxicities are also mentioned. The toxic effects and the potential underlying mechanisms of selected metal and metal oxide nanoparticles are highlighted and described in detail. General factors that affected nanoparticle‐induced toxicity are introduced. [ABSTRACT FROM AUTHOR]

Abstract: Mutations in FLNC for a long time are known in connection to neuromuscular disorders and only recently were described in association with various cardiomyopathies. Here, we report a new clinical phenotype of filaminopathy in four unrelated patients with early‐onset restrictive cardiomyopathy (RCM) in combination with congenital myopathy due to FLNC mutations (NM_001458.4:c.3557C>T, p.A1186V, rs1114167361 in three probands and c.[3547G>C; 3548C>T], p.A1183L, rs1131692185 in one proband). In all cases, concurrent myopathy was confirmed by neurological examination, electromyography, and morphological studies. Three of the patients also presented with arthrogryposis. The pathogenicity of the described missense variants was verified by cellular and morphological studies and by in vivo modeling in zebrafish. Combination of in silico and experimental approaches revealed that FLNC missense variants localized in Ig‐loop segments often lead to development of RCM. The described FLNC mutations associated with early‐onset RCMP extend cardiac spectrum of filaminopathies and facilitate the differential diagnosis of restrictive cardiac phenotype associated with neuromuscular involvement in children. [ABSTRACT FROM AUTHOR]

Crotamine is defensin-like cationic peptide from rattlesnake venom that possesses anticancer, antimicrobial, and antifungal properties. Despite these promising biological activities, toxicity is a major concern associated with the development of venom-derived peptides as therapeutic agents. In the present study, we used zebrafish as a system model to evaluate the toxicity of rhodamine B-conjugated (RhoB) crotamine derivative. The lethal toxic concentration of RhoB-crotamine was as low as 4 μM, which effectively kill zebrafish larvae in less than 10 min. With non-lethal concentrations (<1 μM), crotamine caused malformation in zebrafish embryos, delayed or completely halted hatching, adversely affected embryonic developmental programming, decreased the cardiac functions, and attenuated the swimming distance of zebrafish. The RhoB-crotamine translocated across vitelline membrane and accumulated in zebrafish yolk sac. These results demonstrate the sensitive responsivity of zebrafish to trial crotamine analogues for the development of novel therapeutic peptides with improved safety, bioavailability, and efficacy profiles. [ABSTRACT FROM AUTHOR]

Background: Neural tube defects (NTDs) are among the most common congenital defects affecting approximately 1 in 1000 live births in North America. Their etiology is complex including environmental and genetic factors. Defects in the planar cell polarity (PCP) signaling pathway have been strongly associated with NTDs in animal models and human cohorts. Protein tyrosine kinase 7 (Ptk7) was shown to cause a very severe form of NTDs called craniorachischisis in a mouse model and genetically interacts with a core PCP member Vangl2 where double heterozygotes suffer from spina bifida. In this study, we examined the role of PTK7 in human NTDs to determine whether variants at this gene predispose to these defects. Methods: We sequenced the coding region and the exon-intron junctions of PTK7 in a cohort of 473 patients affected with various forms of open and closed NTDs. Novel and rare variants(<1%) were genotyped in a cohort of 473 individuals. Their pathogenic effect was predicted in silico and functionally in an overexpression assay in a well-established zebrafish model. Results: We identified in our cohort 6 rare variants, 3 of which were absent in public databases. One variant, p.Gly348Ser, acted as a hypermorph when overexpressed in the zebrafish model. Conclusion: We detected potentially pathogenic PTK7 variants in 1.1% of our NTD cohort. Our findings implicate PTK7 as a risk factor for NTDs and provide additional evidence for a pathogenic role of PCP signaling in these malformations. [ABSTRACT FROM AUTHOR]

ABSTRACT MED13L is a component subunit of the Mediator complex, an important regulator of transcription that is highly conserved across eukaryotes. Here, we report MED13L disruption in a translocation t(12;19) breakpoint of a patient with Pierre- Robin syndrome, moderate intellectual disability, craniofacial anomalies, and muscular defects. The phenotype is similar to previously described patients with MED13L haploinsufficiency. Knockdown of MED13L orthologue in zebrafish, med13b, showed early defective migration of cranial neural crest cells ( NCCs) that contributed to cartilage structure deformities in the later stage, recapitulating craniofacial anomalies seen in human patients. Notably, we observed abnormal distribution of developing neurons in different brain regions of med13b morphant embryos, which could be rescued upon introduction of full-length human MED13L m RNA. To compare with mammalian system, we suppressed MED13L expression by short-hairpin RNA in ES-derived human neural progenitors, and differentiated them into neurons. Transcriptome analysis revealed differential expression of components of Wnt and FGF signaling pathways in MED13L-deficient neurons. Our finding provides a novel insight into the mechanism of overlapping phenotypic outcome targeting NCCs derivatives organs in patients with MED13L haploinsufficiency, and emphasizes a clinically recognizable syndromic phenotype in these patients. [ABSTRACT FROM AUTHOR]