MicroRNAs (miRNAs) are small non-coding RNAs that are involved in various biological processes, including apoptosis, by regulating gene expression. This study was designed to test the hypothesis that ethanol-induced downregulation of miR-135a contributes to ethanol-induced apoptosis in neural crest cells (NCCs) by upregulating Siah1 and activating the p38 mitogen-activated protein kinase (MAPK)/p53 pathway. We found that treatment with ethanol resulted in a significant decrease in miR-135a expression in both NCCs and zebrafish embryos. Ethanol-induced downregulation of miR-135a resulted in the upregulation of Siah1 and the activation of the p38 MAPK/p53 pathway and increased apoptosis in NCCs and zebrafish embryos. Ethanol exposure also resulted in growth retardation and developmental defects that are characteristic of fetal alcohol spectrum disorders (FASD) in zebrafish. Overexpression of miRNA-135a significantly reduced ethanol-induced upregulation of Siah1 and the activation of the p38 MAPK/p53 pathway and decreased ethanol-induced apoptosis in NCCs and zebrafish embryos. In addition, ethanol-induced growth retardation and craniofacial defects in zebrafish larvae were dramatically diminished by the microinjection of miRNA-135a mimics. These results demonstrated that ethanol-induced downregulation of miR-135a contributes to ethanol-induced apoptosis in NCCs by upregulating Siah1 and activating the p38 MAPK/p53 pathway and that the overexpression of miRNA-135a can protect against ethanol-induced apoptosis in NCCs and craniofacial defects in a zebrafish model of FASD.

Fetal alcohol spectrum disorders (FASD) are a group of preventable and nongenetic birth defects caused by prenatal alcohol exposure that can result in a range of cognitive, behavioral, emotional, and functioning deficits, as well as craniofacial dysmorphology and other congenital defects. During embryonic development, neural crest cells (NCCs) play a critical role in giving rise to many cell types in the developing embryos, including those in the peripheral nervous system and craniofacial structures. Ethanol exposure during this critical period can have detrimental effects on NCC induction, migration, differentiation, and survival, leading to a broad range of structural and functional abnormalities observed in individuals with FASD. This review article provides an overview of the current knowledge on the detrimental effects of ethanol on NCC induction, migration, differentiation, and survival. The article also examines the molecular mechanisms involved in ethanol-induced NCC dysfunction, such as oxidative stress, altered gene expression, apoptosis, epigenetic modifications, and other signaling pathways. Furthermore, the review highlights potential therapeutic strategies for preventing or mitigating the detrimental effects of ethanol on NCCs and reducing the risk of FASD. Overall, this article offers a comprehensive overview of the current understanding of the impact of ethanol on NCCs and its role in FASD, shedding light on potential avenues for future research and intervention.

Prenatal ethanol exposure is known to cause neurodevelopmental disorders. While high prevalence of epilepsy is observed among the children whose mothers abused alcohol during pregnancy, the results from animal studies are conflicting. Here, we investigated whether embryonic exposure to ethanol can increase the susceptibility to pentylenetetrazole (PTZ)-induced seizures in larval zebrafish. Embryos at 3 hours post-fertilization (hpf) were exposed to ethanol at the concentrations ranging from 0.25% to 1% for 21 hours. Control and ethanol-exposed larvae were challenged with PTZ at 7 days post-fertilization (dpf) at the concentrations of 2.5, 5 or 15 mM. The seizure behavior of larvae was recorded and analyzed using EthoVision XT 11. We found that embryonic ethanol exposure increased the percentage of larvae exhibiting typical stage II and III seizure and resulted in a significant reduction in stage I, II and III seizure latency in an ethanol concentration-dependent manner. Embryonic exposure to ethanol also significantly increased the severity of PTZ-induced seizures in larvae, as demonstrated by increased total distance traveled and the duration of mobility. This is the first demonstration that ethanol exposure during early embryonic stage can reduce the threshold for chemically induced seizures and increase the severity of seizure behavior in larval fish.