Excess ethanol drinking after prenatal exposure to ethanol: chemokine signaling and orexigenic neuropeptides

产前接触乙醇后过量饮酒：趋化因子信号传导和促食欲神经肽

• 批准号: 9082977
• 负责人: SARAH F LEIBOWITZ
• 金额: $ 38.14万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-10 至 2021-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9082977
• 关键词: Adolescence; Adolescent; Adolescent Behavior; Adult Children; Alcohol abuse; Alcohol consumption; Alcoholism; Animals; Anxiety; Appetite Stimulants; Behavior; Birth; Brain; CCL2 gene; Cells; Clinical; Clinical Research; Consumption; Development; Dose; Embryo; Embryonic Development; Ethanol; Exposure to; Fetal Alcohol Exposure; Foundations; Gene Mutation; Genes; Hypothalamic structure; Immune system; Inflammatory; Injection of therapeutic agent; Intake; Label; Lateral; Life; Link; Lipopolysaccharides; Mediating; Mental Depression; Messenger RNA; Modeling; Molecular; Mutation; Neuroepithelial; Neuroglia; Neurons; Neuropeptides; Peptides; Peripheral; Population; Pregnancy; Radial; Rattus; Reporting; Research; Risk; Role; Signal Transduction; Small Interfering RNA; Source; Substance abuse problem; System; Testing; Third ventricle structure; Ventricular; adolescent offspring; alcohol consumption during pregnancy; alcohol effect; alcohol exposure; alcohol response; base; chemokine; density; emotional behavior; fetal; hypocretin; in utero; melanin-concentrating hormone; migration; monocyte chemoattractant protein 1 receptor; nerve stem cell; neuroepithelium; neurogenesis; neuron development; neutralizing antibody; offspring; preference; prenatal; prenatal exposure; public health relevance; research study; tool; underage drinking

 DESCRIPTION (provided by applicant): Clinical studies demonstrate that exposure to ethanol early in life, particularly during gestation, increases the consumption of and preference for ethanol and the risk for alcoholism. The molecular and cellular mechanisms involved in this phenomenon have yet to be characterized. Our recent studies in rats, focusing on the hypothalamic orexigenic neuropeptides, melanin-concentrating hormone (MCH) and orexin (OX), that increase ethanol intake and related emotional behaviors, demonstrate a stimulatory effect of embryonic exposure to ethanol, at relatively low levels and only during peak hypothalamic neurogenesis, on the density and long-term expression of neurons that produce these peptides in the perifornical lateral hypothalamus (PF+LH). Building on evidence that the inflammatory agent lipopolysaccharide, like ethanol, increases peripheral chemokine levels, peptide expression, and drinking of ethanol, we investigated and demonstrated in our most recent study a close link between these orexigenic peptides and local chemokine system, CCL2, and its receptor, CCR2. Consistent with evidence that mutations of CCL2 or CCR2 genes markedly reduce ethanol intake, we found that prenatal ethanol exposure, at low doses from embryonic day 10 (E10) to E15, increases circulating CCL2 levels, density in pre-pubertal offspring of MCH neurons that contain CCR2 but not CCL2, and intake of and preference for ethanol during adolescence. This evidence, along with our extensive preliminary findings, provides the foundation for our hypothesis, to be tested in 4 aims, that: Activation of the CCL2/CCR2 signaling system in the embryo as well as dam contributes to the stimulatory effects of prenatal ethanol exposure, at low levels and for a brief period, on development and migration of orexigenic peptide neurons in the PF+LH, resulting in their increased density and expression after birth that subsequently promote ethanol intake and related behaviors during adolescence. In Aim 1, we will examine in adolescent offspring whether prenatal exposure to CCL2 acts similarly to ethanol and mediates its stimulatory effects on neuronal development and behaviors induced by the peptides. In Aim 2, we will directly compare in the embryo effects of prenatal exposure to ethanol and CCL2 and determine whether CCL2 mediates ethanol-induced proliferation, differentiation and migration of peptide neurons that co-express CCR2. Building on our preliminary finding that radial glia in the hypothalamic neuroepithelium (NEP) contain CCL2 and are highly responsive to ethanol, we propose in Aim 3 to test in the embryo, from E12 to E19, the effects of ethanol or CCL2 on development of these neuroprogenitor cells and peptide neurons in NEP and the role of these CCL2 cells in mediating ethanol's effects. In Aim 4, we will test, with direct manipulations of the embryo involving third ventricular injections of CCL2 or CCL2 siRNA, whether neuroepithelial CCL2 is involved in ethanol's effects on neuronal development and adolescent drinking. This research should provide fundamentally new information on how gestational exposure to ethanol acts through the neuroimmune system to control the development of peptide neurons that contribute to substance abuse.

 描述（由申请人提供）：临床研究表明，在生命早期暴露于乙醇，特别是在妊娠期间，会增加乙醇的消耗和偏好以及酒精中毒的风险。这一现象所涉及的分子和细胞机制尚待确定。我们最近对大鼠的研究，重点是下丘脑食欲神经肽，黑色素浓缩激素（MCH）和食欲素（OX），增加乙醇摄入量和相关的情绪行为，证明胚胎暴露于乙醇的刺激作用，在相对低的水平，只有在高峰期下丘脑神经发生，对穹窿周围外侧下丘脑（PF+LH）中产生这些肽的神经元的密度和长期表达的影响。基于炎症因子脂多糖（如乙醇）增加外周趋化因子水平、肽表达和饮用乙醇的证据，我们在最近的研究中调查并证明了这些食欲肽与局部趋化因子系统CCL 2及其受体CCR 2之间的密切联系。与CCL 2或CCR 2基因突变显著降低乙醇摄入量的证据一致，我们发现，产前乙醇暴露，在胚胎第10天（E10）至E15的低剂量下，增加循环CCL 2水平，MCH神经元的青春期前后代的密度，其中含有CCR 2但不含CCL 2，以及青春期期间乙醇的摄入和偏好。这一证据，沿着我们广泛的初步发现，为我们的假设提供了基础，将在4个目标中进行测试，即：胚胎和母体中CCL 2/CCR 2信号系统的激活有助于产前低水平和短暂乙醇暴露对PF+LH中食欲肽神经元的发育和迁移的刺激作用，导致出生后其密度和表达增加，随后促进青春期的乙醇摄入和相关行为。在目标1中，我们将在青少年后代中研究产前暴露于CCL 2是否与乙醇类似，并介导其对肽诱导的神经元发育和行为的刺激作用。在目标2中，我们将直接比较胎儿期暴露于乙醇和CCL 2的影响，并确定CCL 2是否介导乙醇诱导的共表达CCR 2的肽神经元的增殖、分化和迁移。基于我们的初步发现，放射状胶质细胞在下丘脑神经上皮细胞（NEP）含有CCL 2，并高度响应于乙醇，我们建议在目标3测试胚胎，从E12到E19，乙醇或CCL 2对这些神经祖细胞和肽神经元在NEP的发展和这些CCL 2细胞在介导乙醇的作用的作用。在目标4中，我们将测试，直接操作胚胎，包括第三脑室注射， CCL 2或CCL 2 siRNA，神经上皮CCL 2是否参与乙醇对神经元发育和青少年饮酒的影响。这项研究应该提供关于妊娠期暴露于乙醇如何通过神经免疫系统控制导致药物滥用的肽神经元发育的新信息。