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令人严重关切的问题 由于这些药物的有害影响， 对母亲和胎儿产生的影响。 本项目的目标 目的是研究胎盘在并发症发病机制中的作用 滥用可卡因和安非他明引起的精神疾病和 确定胎盘是这些药物直接靶点。 人胎盘 表达血清素转运体和去甲肾上腺素转运体， 其中可卡因和/或安非他明的目标。 这些转运蛋白 最可能的功能是清除血管活性单胺血清素 和去甲肾上腺素 干涉他们的 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功能， 调节和胎盘中这两种转运蛋白的个体发育。 研究 将包括可卡因和 安非他命和这两种转运蛋白 这些研究将与 分离的质膜囊泡从胎盘，也与完整的 滋养层细胞和绒毛膜癌细胞。 胎盘 合体滋养层是极化细胞，免疫定位研究将 以确定血清素的表达是否 和去甲肾上腺素转运蛋白是极化的， 面向母体的刷状缘膜和面向胎儿的基底膜。 拟议中的研究还将描述以下方面的不同作用： 激素/细胞因子在这些转运蛋白的调节中的作用 滋养层培养物和绒毛膜癌细胞。 这些研究将包括 分析激素/精氨酸诱导的转运活性变化， 转运蛋白密度和转运蛋白特异性mRNA的稳态水平。 介导激素/细胞因子效应的特定信号通路将 并参与转录和翻译 过程和翻译后修饰的调控将是 描绘的。 研究还将验证这一假设，即受损的功能， 这些转运蛋白中的一种或两种是先兆子痫的致病因素， 临床上在许多方面类似于并发症的情况 怀孕期间使用可卡因和安非他明引起的。 的 5-羟色胺和去甲肾上腺素转运体的个体发育 将使用大鼠作为实验模型研究胎盘。

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.