CELLULAR TARGETS FOR ABUSABLE DRUGS IN HUMAN PLACENTA

描述：(申请人摘要) 怀孕期间滥用非法药物目前是一个严重的问题。 与妇女的健康有关，因为这些药物有有害的影响 对母亲和胎儿都有影响。当前项目的目标 探讨胎盘在并发症发病机制中的作用 因在怀孕期间滥用可卡因和安非他明而引起的 确定胎盘是这些药物的直接靶点。人胎盘 表达5-羟色胺转运体和去甲肾上腺素转运体，两者都是 其中一些是可卡因和/或安非他明的目标。这些传送器 最有可能在清除血管活性单胺类5-羟色胺中起作用 以及绒毛间隙中的去甲肾上腺素。干扰他们的 可卡因和安非他明的作用会对 发育中的胎儿。目前的项目将调查正常的功能， 胎盘中这两种转运蛋白的调控和个体发育。研究 将包括可卡因和可卡因相互作用的详细特征 安非他命和这两个传送器。这些研究将在 从胎盘分离的质膜囊泡和完整的 滋养层细胞和绒毛膜癌细胞。胎盘 合体滋养层细胞是一种极化细胞，免疫定位研究将 以确定是否有5-羟色胺的表达 去甲肾上腺素转运体是两极化的 面向母体的刷状缘膜和面向胎儿的基底膜。 拟议的研究还将描绘出不同的作用 激素/细胞因子在这些转运蛋白的调节中使用初级 滋养层细胞培养和绒毛膜癌细胞。这些研究将包括 分析激素/细胞因子引起的转运活性的变化， 转运蛋白密度和转运蛋白特异性mRNAs的稳态水平。 介导激素/细胞因子效应的特定信号通路将 被识别以及转录和翻译的参与 法规中的流程和翻译后修改将是 勾勒出来的。研究还将检验这样一种假设，即大脑功能受损 这些转运蛋白中的一个或两个都是先兆子痫的致病因素， 临床上在许多方面与并发症相似的情况 由怀孕期间使用可卡因和安非他明引起的。这个 5-羟色胺和去甲肾上腺素转运体的个体发育 胎盘将以大鼠为实验模型进行研究。

DESCRIPTION: (Applicant's Abstract) Abuse of illicit drugs during pregnancy is currently a serious concern pertaining to women's health because of the deleterious effects these drugs have on the mother and on the fetus. The objective of the current project is to investigate the role of placenta in the pathogenesis of complications arising from abuse of cocaine and amphetamines during pregnancy and to establish that placenta is a direct target for these drugs. Human placenta expresses the serotonin transporter and the norepinephrine transporter, both of which are targets for cocaine and/or amphetamines. These transporters most likely function in the clearance of the vasoactive monoamines serotonin and norepinephrine from the intervillous space. Interference with their functions by cocaine and amphetamines would have had effects on the developing fetus. The current project will investigate the normal function, regulation, and ontogeny of these two transporters in the placenta. Studies will include detailed characterization of the interaction of cocaine and amphetamines with these two transporters. These studies will be done with isolated plasma membrane vesicles from the placenta and also with intact trophoblast cells and choriocarcinoma cells. The placental syncytiotrophoblast is a polarized cell and immunolocalization studies will be performed to determined whether or not the expression of the serotonin and norepinephrine transporters is polarized with regard to the maternal-facing brush border membrane and the fetal-facing basal membrane. Proposed studies will also delineate the differential role of hormones/cytokines in the modulation of these transporters using primary trophoblast cultures and choriocarcinoma cells. These studies will include analysis of hormone/cytokine-induced changes in the transport activity, transporter density and steady state levels of transporter-specific mRNAs. The specific signalling pathways mediating the hormone/cytokine effects will be identified and the involvement of transcriptional and translational processes and posttranslational modifications in the regulation will be delineated. Studies will also test the hypothesis that impaired function of one or both of these transporters is a pathogenic factor in preeclampsia, a condition which clinically resembles in many respects the complications arising from use of cocaine and amphetamines during pregnancy. The ontogenic development of the serotonin and norepinephrine transporters in placenta will be studied using the rat as the experimental model.