CONTROL AND INTEGRATION OF GNRH NEURAL PATHWAYS

GNRH 神经通路的控制和整合

• 批准号: 3311445
• 负责人: PAUL C GOLDSMITH
• 金额: $ 11.38万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 1977
• 资助国家: 美国
• 起止时间: 1977-04-01 至 1986-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3311445
• 关键词: catecholamines; dendrites; dopamine; electron microscopy; epinephrine; gonadotropin releasing factor; haloperidol; histochemistry /cytochemistry; immunochemistry; immunoperoxidase; median eminence; neuroendocrine system; sectioning; sex behavior disorder; sex development disorder; silver impregnation; stainings; synapses; synaptosomes; tyrosine 3 monooxygenase

Gonadotropin releasing hormone (GnRH) containing neurons in the primate medial basal hypothalamus (MBH), and their relationship to other peptidergic, catecholaminergic, and estrogen-synthesizing neurons in the MBH and median eminence (ME) of the monkey and rat are investigated. 1. Apparent contacts between GnRH neurons in the baboon MBH and its ventral hypothalamic tract (VHT) are identified by immunocytochemical (ICC) staining on vibratome sections, observed with light microscopy (LM), and then examined with electron microscopy (EM). Evidence for electrotonic (gap) junctions between MBH neurons is obtained using retrograde Lucifer Yellow (LY) labelling in the stalk-sectioned rhesus monkey. LY fluorescent, dye-coupled cells are ICC stained for GnRH using the PAP technique, and examined by EM for gap junctions. Other evidence for gap junctions is sought using freeze fracture of the primate VHT. 2) Cell bodies of origin of the VHT and tuberoinfundibular pathways are identified with retrograde True Blue (TB) labelling in the stalksectioned rhesus monkey. TB fluorescent cells are stained by ICC to determine the GnRH component. Results with TB are compared to those with LY, and the cell body distribution mapped. 3) Localization of aromatase (ARO) in the rat and baboon brain is determined using ICC for specific ARO enzyme subunits at the LM level. Special attention is given to the medial preoptic area, the location of the sexually dimorphic nucleus (SDN-POA) in rats, and the MBH, the two brain regions with highest ARO activity. If the rat SDN-POA is ARO-positive, we will ICC stain for ARO to identify the equivalent of the SDN-POA in the baboon. 4) Relationships between receptors for dopamine (DA), b-endorphin (END), and prolactin (PRL), and the ME nerve terminals on which they occur is investigated. Using ME synaptosomes, DA, END, PRL receptors are identified with ICC and GaRIgG-colloidal gold. The content of labelled synaptosomes is determined using ICC for GnRH, END, or tyrosine hydroxylasle (TH, as a marker for DA neurons) and the PAP technique. 5) Interactions between GnRH neurons and others belived to affect GnRH secretion are examined in the rat and baboon MBH. Dual-label ICC is performed for GnRH followed by ICC for END, ARO, or TH. Interactions identified by LM are examined by EM. This information regarding GnRH neuronal systems, interconnections, and afferent input will enhance our knowledge and understanding of neural control of reproduction in primates, including humans.

灵长类动物中含有促性腺激素释放激素（GnRH）的神经元 内侧基底下丘脑（MBH），以及它们与其他 肽能、儿茶酚胺能和雌激素合成神经元 对猴和大鼠的正中隆起（ME）和正中高（MBH）进行了研究。 1. 狒狒MBH及其腹侧的GnRH神经元之间的表观接触 免疫细胞化学（ICC）鉴定下丘脑束（VHT） 在振动切片上染色，用光学显微镜（LM）观察，和 然后用电子显微镜（EM）检查。 电紧张性证据 (gap)使用逆行Lucifer获得MBH神经元之间的连接 黄色（LY）标记的茎切片恒河猴。 LY 使用PAP对荧光染料偶联的细胞进行GnRH ICC染色 技术，并通过EM检查间隙连接。 差距的其他证据 使用灵长类动物VHT的冷冻断裂来寻找连接。 2)细胞 VHT和结节漏斗径路的起源体被识别 用真蓝（TB）逆行标记法， 猴子. TB荧光细胞用ICC染色以测定GnRH 成分 将TB的结果与LY的结果进行比较， 身体分布图 3)芳香化酶（ARO）在大鼠体内的定位 而狒狒脑则用ICC测定特定的ARO酶亚基 LM级别。 特别注意内侧视前区， 大鼠性二形核（SDN-POA）的位置， MBH是ARO活性最高的两个大脑区域。 如果大鼠SDN-POA 是ARO阳性，我们将ICC染色的ARO，以确定相当于 狒狒体内的SDN-POA 4)多巴胺受体之间的关系 (DA)，b-内啡肽（END）和催乳素（PRL），以及ME神经末梢 他们所发生的事情正在调查中。 使用ME突触体、DA、END、PRL 用ICC和GaRIgG-胶体金鉴定受体。 内容 标记的突触体的数量使用针对GnRH、END或酪氨酸的ICC来确定。 羟化酶（TH，作为DA神经元的标记物）和PAP技术。 第五章） GnRH神经元和其他被认为影响GnRH的神经元之间的相互作用 在大鼠和狒狒MBH中检查分泌。 双标签ICC是 进行GnRH检查，然后进行END、ARO或TH的ICC检查。 相互作用 由LM识别的，由EM检查。 关于GnRH的信息 神经元系统、相互连接和传入输入将增强我们的 对灵长类动物生殖的神经控制的认识和理解， 包括人类