CHARACTERIZATION OF HORMONE CONCENTRATING NEURONS

许多性腺激素依赖的神经内分泌事件和行为 被认为是由类固醇激素通过视前症起作用的， 含有类固醇特异性的下丘脑和边缘系统神经元 受体蛋白。这项提案中的实验旨在 在逐个细胞的基础上(IA)显示下丘脑外轴突 肾上腺皮质类固醇激素集中神经元的联系 类固醇放射自显影结合逆行神经解剖示踪。 这些实验将准确地揭示集中类固醇的神经元如何适应 在行为相关神经回路的神经内分泌学中。其他 实验(IB)旨在检测化学成分， 性腺类固醇的神经递质或神经肽相关 激素集中的神经元，也是以细胞为基础，使用类固醇 放射自显影结合免疫细胞化学。这些神经元可能是 性腺类固醇激素影响的部分机制 神经递质或神经肽对下丘脑释放的调节 荷尔蒙，随后影响脑下垂体前叶的分泌。 实验(II)旨在检查神经内分泌特性 POA或下丘脑内侧基底核(MBH)的内源性神经元 确定类固醇集中能力，或神经递质或 手术隔离岛后存活的神经细胞的神经肽含量 POA或MBH。内源性下丘脑的生理学意义 神经元可能是相当多的；对其属性的逐个细胞分析 还没有实施。 类固醇激素对含有特定基因的细胞基因组的作用 类固醇激素受体是最有证据的机制，通过它 类固醇会影响细胞。旨在优化使用的实验(III) 与cdna探针在脑细胞上的原位杂交将是 使用探针1)催产素；2)加压素和3) 酪氨酸羟基酶。然后，这些程序将在一个范例中使用 旨在定性和定量地证明 生理状态或类固醇激素对下丘脑mRNA水平的影响 用cDNAs探针进行原位检测。有限的尝试(IV) 用免疫组织化学方法分析下丘脑神经元雌二醇受体(E-R)含量 使用E-R初级抗体的免疫细胞化学方法将是 被执行。所有这些实验的长期目标是揭示，在 以细胞为基础，性腺的特点。

Many gonadal-steroid dependent neuroendocrine events and behaviors are thought to be mediated by steroid hormones acting through preoptic, hypothalamic and limbic system neurons which contain steroid-specific receptor proteins. The experiments in this proposal are designed to reveal, on a cell-by-cell basis (IA) the extrahypothalamic axonal connections of gonadal steroid hormone-concentrating neurons by using steroid autoradiography combined with retrograde neuroanatomical tracing. These experiments will reveal exactly how steroid concentrating neurons fit in neuroendocrinologically of behaviorally relevant neural circuits. Other experiments (IB) are designed to detect the chemical content, neurotransmitter or neuropeptide-related, of gonadal steroid hormone-concentrating neurons, also on a cell-by-cell basis, using steroid autoradioagraphy combined with immunocytochemistry. These neurons could be part of the mechanism by which gonadal steroid hormones influence neurotransmitter or neuropeptide regulation of hypothalamic releasing hormones, which subsequently effect anterior pituitary secretion. Experiments (II) designed to examine the neuroendocrine properties of intrinsic neurons of the POA or medial basal hypothalamus (MBH) will identify steroid-concentrating capacity, or neurotransmitter or neuropeptide content, of neurons that survive surgical isolation of islands of POA or MBH. The physiologic importance of the intrinsic hypothalamic neurons may be considerable; cell-by-cell analysis of their properties has yet to be carried out. The action of steroid hormones on the genome of cells that contain specific steroid hormone receptors is the best documented mechanism by which steroids influence cells. Experiments (III) designed to optimize the use of in situ hybridization with cDNA probes on cells of the brain will be carried out using probes to 1) prooxytocin; 2) provasopressin and 3) tyrosine hydroxylase. Then, these procedures will be used in a paradigm designed to demonstrate qualitatively and quantitatively the effects of physiological state or steroid hormones on mRNA levels of hypothalamic neurons, detected in situ with the cDNA probes. Limited attempts (IV) to analyze the estradiol receptor (E-R) content of hypothalamic neurons with immunocytochemical methods using primary antibodies to the E-R will be carried out. The long term goal of all these experiments is to reveal, on a cell-by-cell basis, the characteristics of gonadal.