CHARACTERIZATION OF HORMONE CONCENTRATING NEURONS: THEI

许多性腺类固醇依赖性神经内分泌事件和行为是 被认为是由类固醇激素通过视前区起作用介导的， 下丘脑和边缘系统神经元含有类固醇特异性 受体蛋白 本提案中的实验旨在 揭示，在一个细胞的基础上（IA）下丘脑外轴突 连接的性腺类固醇激素集中神经元，通过使用 类固醇放射自显影结合逆行神经解剖追踪。 这些实验将揭示类固醇集中神经元如何适应 行为相关神经回路的研究。 其他 设计实验（IB）以检测化学成分， 神经递质或神经肽相关的、性腺类固醇 也是一个细胞一个细胞地使用类固醇 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 下丘脑释放的神经递质或神经肽调节 激素，随后影响垂体前叶分泌。 实验（II）旨在检查神经内分泌特性的 POA或内侧基底下丘脑（MBH）的内在神经元将 确定类固醇浓缩能力，或神经递质或 神经肽含量，神经元存活的岛屿手术隔离 或MBH。 下丘脑固有神经元的生理学意义 神经元可能是相当大的;细胞对它们的属性进行分析， 还有待执行。 类固醇激素对含有特异性 类固醇激素受体是最好的记录机制， 类固醇影响细胞。 实验（三）优化使用设计 用cDNA探针对脑细胞进行原位杂交， 使用针对1）催产素原; 2）加压素原和3） 酪氨酸羟化酶 然后，这些程序将被用于一个范例 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 神经元，原位检测的cDNA探针。 有限的尝试（四） 分析下丘脑神经元雌二醇受体（E-R）含量， 使用E-R的一级抗体的免疫细胞化学方法将是 贯彻 所有这些实验的长期目标是揭示， 一个细胞一个细胞的基础上，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.