Prenatal Alcohol Exposure Inhibits Transient Expression of Autophagy and Synaptic Proteins in Developing Brain.

Obstetrics and gynecology research Pub Date : 2025-01-01 Epub Date: 2025-01-22 DOI:10.26502/ogr0173
Monica Hampe, Nune Darbinian, Nana Merabova, Armine Darbinyan, Jamal Hamze, Uday Bharai, Yuri Persidsky, Mary F Morrison, Shohreh Amini, Laura Goetzl, Michael E Selzer
{"title":"Prenatal Alcohol Exposure Inhibits Transient Expression of Autophagy and Synaptic Proteins in Developing Brain.","authors":"Monica Hampe, Nune Darbinian, Nana Merabova, Armine Darbinyan, Jamal Hamze, Uday Bharai, Yuri Persidsky, Mary F Morrison, Shohreh Amini, Laura Goetzl, Michael E Selzer","doi":"10.26502/ogr0173","DOIUrl":null,"url":null,"abstract":"<p><strong>Introduction: </strong>Neuronal apoptosis and consequent inhibition of autophagy, with loss of synaptic connections are central events in the genesis of fetal alcohol spectrum disorders (FASD). However, studies of molecular mechanisms of autophagy in human fetal brain are limited. Recently, prenatal exposure to EtOH was associated with reduced miRNA-9 levels in fetal brain-derived exosomes (FB- Es) isolated from maternal plasma, which correlated with small eyes, an anatomical hallmark of fetal alcohol syndrome (FAS). Since miR-9 targets several genes that regulate synaptic plasticity, EtOH-induced inhibition of miR-9 could potentially result in dysregulation of genes involved in synaptogenesis/plasticity.</p><p><strong>Methods: </strong>Rats were fed a 6.7% EtOH liquid diet from E16 to birth. Human studies: Fetal brain tissues from elective pregnancy terminations were collected at 9-23 weeks gestational age (GA; n=20 EtOH-exposed and 20 GA- and fetal sex-matched unexposed controls). EtOH consumption was assessed by questionnaire (adapted from NIAAA PASS). Expressions of 84 genes in a synaptic plasticity microarray were assessed in human fetal brain samples, verified by qRT-PCR, and for some mRNAs, copy number was determined in FB-Es by droplet digital PCR. Protein expression was measured in brain by qWestern blot assays or with a MAP kinase microarray. Exosomal protein levels were measured by ELISA.</p><p><strong>Results: </strong>Levels of pro-apoptotic caspase-3 and Bax were significantly increased in the brains of EtOH-exposed rat pups, while early expressions of anti-apoptotic Bcl2 and biphasic Bag3 were inhibited. Phosphorylation of GSK3β was increased, and during Bag3 inhibition, the GSK3β target β-catenin also was increased. EtOH-exposed P8 and P15 rats showed motor abnormalities during low Bag3 expression. EtOH exposure reduced expression of synaptophysin and synapsin. In most synaptic plasticity pathways, levels of mRNAs were reduced. Several immediate-early genes were upregulated, but SYNPO, which is involved late in synaptic plasticity was downregulated 78%. Genes involved in Long Term Potentiation (LTP) and long-term depression (LTD) were downregulated, but the neurotoxic cytokine TNF⍺ was upregulated.</p><p><strong>Conclusions: </strong>Prenatal exposure to EtOH was associated with reduced expression of autophagy genes in the fetal brains of rats and humans. Inhibition of Bag3 correlated with upregulation of GSK3β and its downstream targets, suggesting dysregulation of β-catenin signaling. Synaptic proteins, including those implicated in LTP and LTD also were inhibited by EtOH. The results in FB-E mirrored those in brain tissue. Reduced expression of miR-9 target synaptic genes in FB-Es might serve as novel biomarkers to predict FASD.</p>","PeriodicalId":74336,"journal":{"name":"Obstetrics and gynecology research","volume":"8 1","pages":"10-24"},"PeriodicalIF":0.0000,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11905422/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Obstetrics and gynecology research","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.26502/ogr0173","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2025/1/22 0:00:00","PubModel":"Epub","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0

Abstract

Introduction: Neuronal apoptosis and consequent inhibition of autophagy, with loss of synaptic connections are central events in the genesis of fetal alcohol spectrum disorders (FASD). However, studies of molecular mechanisms of autophagy in human fetal brain are limited. Recently, prenatal exposure to EtOH was associated with reduced miRNA-9 levels in fetal brain-derived exosomes (FB- Es) isolated from maternal plasma, which correlated with small eyes, an anatomical hallmark of fetal alcohol syndrome (FAS). Since miR-9 targets several genes that regulate synaptic plasticity, EtOH-induced inhibition of miR-9 could potentially result in dysregulation of genes involved in synaptogenesis/plasticity.

Methods: Rats were fed a 6.7% EtOH liquid diet from E16 to birth. Human studies: Fetal brain tissues from elective pregnancy terminations were collected at 9-23 weeks gestational age (GA; n=20 EtOH-exposed and 20 GA- and fetal sex-matched unexposed controls). EtOH consumption was assessed by questionnaire (adapted from NIAAA PASS). Expressions of 84 genes in a synaptic plasticity microarray were assessed in human fetal brain samples, verified by qRT-PCR, and for some mRNAs, copy number was determined in FB-Es by droplet digital PCR. Protein expression was measured in brain by qWestern blot assays or with a MAP kinase microarray. Exosomal protein levels were measured by ELISA.

Results: Levels of pro-apoptotic caspase-3 and Bax were significantly increased in the brains of EtOH-exposed rat pups, while early expressions of anti-apoptotic Bcl2 and biphasic Bag3 were inhibited. Phosphorylation of GSK3β was increased, and during Bag3 inhibition, the GSK3β target β-catenin also was increased. EtOH-exposed P8 and P15 rats showed motor abnormalities during low Bag3 expression. EtOH exposure reduced expression of synaptophysin and synapsin. In most synaptic plasticity pathways, levels of mRNAs were reduced. Several immediate-early genes were upregulated, but SYNPO, which is involved late in synaptic plasticity was downregulated 78%. Genes involved in Long Term Potentiation (LTP) and long-term depression (LTD) were downregulated, but the neurotoxic cytokine TNF⍺ was upregulated.

Conclusions: Prenatal exposure to EtOH was associated with reduced expression of autophagy genes in the fetal brains of rats and humans. Inhibition of Bag3 correlated with upregulation of GSK3β and its downstream targets, suggesting dysregulation of β-catenin signaling. Synaptic proteins, including those implicated in LTP and LTD also were inhibited by EtOH. The results in FB-E mirrored those in brain tissue. Reduced expression of miR-9 target synaptic genes in FB-Es might serve as novel biomarkers to predict FASD.

产前酒精暴露抑制发育中的大脑自噬和突触蛋白的瞬时表达。
导读:神经元凋亡和随之而来的自噬抑制,以及突触连接的丧失是胎儿酒精谱系障碍(FASD)发生的中心事件。然而,对人类胎儿大脑自噬的分子机制研究有限。最近,产前暴露于EtOH与从母体血浆中分离的胎儿脑源性外泌体(FB- Es)中miRNA-9水平降低有关,这与小眼睛有关,小眼睛是胎儿酒精综合征(FAS)的解剖学标志。由于miR-9靶向调节突触可塑性的几个基因,因此etoh诱导的miR-9抑制可能潜在地导致参与突触发生/可塑性的基因失调。方法:大鼠从16岁起至出生时饲喂含6.7%乙醇的液体饲料。人类研究:收集9-23周胎龄(GA;n=20例etoh暴露组和20例GA和胎儿性别匹配的未暴露组)。采用问卷(改编自NIAAA PASS)评估EtOH消耗。在人胎儿脑样本中,我们评估了突触可塑性芯片中84个基因的表达,并通过qRT-PCR进行了验证,对一些mrna,我们用微滴数字PCR测定了FB-Es中的拷贝数。用western blot或MAP激酶芯片检测脑内蛋白表达。ELISA法检测外泌体蛋白水平。结果:etoh暴露大鼠脑内促凋亡caspase-3和Bax水平显著升高,抗凋亡Bcl2和双相Bag3的早期表达受到抑制。GSK3β磷酸化增加,在Bag3抑制期间,GSK3β靶蛋白β-catenin也增加。etoh暴露的P8和P15大鼠在Bag3低表达时出现运动异常。EtOH降低了synaptophysin和synapsin的表达。在大多数突触可塑性通路中,mrna水平降低。几个直接早期基因上调,但参与突触可塑性晚期的SYNPO基因下调78%。参与长期增强(LTP)和长期抑制(LTD)的基因下调,但神经毒性细胞因子TNF上调。结论:产前暴露于EtOH与大鼠和人类胎儿大脑中自噬基因表达降低有关。Bag3的抑制与GSK3β及其下游靶点的上调相关,提示β-catenin信号通路失调。突触蛋白,包括LTP和LTD的相关蛋白也被EtOH抑制。FB-E的结果反映了脑组织的结果。FB-Es中miR-9靶突触基因的表达降低可能作为预测FASD的新生物标志物。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
自引率
0.00%
发文量
0
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信