CONTROL AND INTEGRATION OF GNRH NEURAL PATHWAYS

GNRH 神经通路的控制和整合

• 批准号: 2196756
• 负责人: PAUL C GOLDSMITH
• 金额: $ 24.04万
• 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 1996-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2196756
• 关键词: Macaca fascicularis; afferent nerve; animal puberty; dopamine; endorphins; estrogen receptors; excitatory aminoacid; gamma aminobutyrate; gonadotropin releasing factor; hormone regulation /control mechanism; immunocytochemistry; immunoelectron microscopy; median eminence; neural information processing; neuroanatomy; neuroendocrine system; neurotransmitters; norepinephrine; peroxidases; pituitary gonadal axis; progesterone receptors; serotonin; synapses

Scattered GnRH neurons in the primate hypothalamus perform different functions and engage in selective interactions. To establish which interactions are relevant to neuroendocrine regulation, (1) We will identify neuroendocrine GnRH neurons in the primate, and determine the afferent inputs involved in their control. We will identify neuroendocrine GnRH neurons in pre-ovulatory female primates by retrograde labeling from the median eminence (ME) and immunocytochemical (ICC) staining for GnRH with. DAB. We will use ICC staining with colloidal gold (red) to identify their specific afferents, e.g. dopamine (DA), norepinephrine (NE), and serotonin (5-HT), the peptide beta-endorphin (b-E), gamma-aminobutyric acid (GABA), and the excitatory amino acids (EAAs) glutamate and aspartate. Synapses will be characterized at the electron microscopic (EM) level and quantified. (2) We will determine which afferents mediate estrogen (E2) and progesterone (P4) feedback on neuroendocrine GnRH neurons. Since GnRH neurons lack E2 receptors, inhibitory feedback is thought to occur through afferent DA, b-E, or GABA neurons, which concentrate E2 and in some cases P4. We will use ICC staining with heavy metal-intensified DAB (black) to reveal neuronal nuclei containing E2 receptors in pre-ovulatory female primates, and identify neuroendocrine GnRH neurons with standard DAB (brown). We will perform ICC staining for DA, b-E, or GABA neurons with colloidal gold to identify which afferents mediate E2 feedback. Types and numbers of synapses will be evaluated by EM and quantified. Using the same protocol, we will perform ICC staining for P4 receptors and determine which afferents mediate P4 feedback. We will repeat these studies in early follicular and mid-luteal phase animals to examine feedback sites and effects of changing steroid levels. Puberty in primates involves reactivation of the dormant prepubertal GnRH pulse generator. While this implies that a competent GnRH neuronal system is present in juveniles, neuroanatomical changes which ma\, enhance pulsatile GnRH secretion have yet to be assessed. (3) We will determine if GnRH neuronal connections or afferent synapses are altered during the peripubertal period. We will perform double EM ICC staining for GnRH. neurons (with DAB) and their afferents (with colloidal gold) in prepubertal and adult male monkeys. We will quantify and compare the types, characteristics, and numbers of GnRH neuronal interactions and afferent synapses. Using serial plasma LH measurements (by RIA), we will identify peripubertal (nocturnal LH surge) animals, perform ICC staining, and evaluate participation of the neural changes observed. In subsequent studies, we will examine prepubertal and peripubertal female monkeys and quantify their GnRH neurointeractions. We will compare the data to that previously obtained in adult, cycling females, and correlate the observed changes with associated peripubertal events. Together, these studies will elucidate the nature of GnRH neuronal control, the generation of pulsatile GnRH secretion, and assess neuroanatomical changes contributing to the onset of puberty.

灵长类动物下丘脑中分散的GnRH神经元执行不同的 功能，并参与选择性互动。 确定哪些 相互作用与神经内分泌调节有关，（1）我们将 识别灵长类动物中的神经内分泌GnRH神经元，并确定 参与其控制的传入输入。 我们将确定 排卵前雌性灵长类动物的神经内分泌GnRH神经元 正中隆起逆行标记和免疫细胞化学 (ICC)GnRH染色。 民建联 我们将使用ICC染色， 胶体金（红色），以确定其特定的传入，如多巴胺 (DA)去甲肾上腺素（NE）和5-羟色胺（5-HT）， β-内啡肽（b-E）、γ-氨基丁酸（GABA）和兴奋性 氨基酸（EAA）谷氨酸和天冬氨酸。 突触将是 在电子显微镜（EM）水平表征并定量。（二） 我们将确定哪些传入介导雌激素（E2）和孕激素 (P4)反馈给神经内分泌GnRH神经元。 由于GnRH神经元缺乏E2， 受体，抑制反馈被认为是通过传入DA发生， b-E或GABA神经元，其浓缩E2，在某些情况下浓缩P4。 我们将 用重金属增强的DAB（黑色）进行ICC染色， 排卵前雌性灵长类动物中含有E2受体的神经元核， 用标准DAB（棕色）鉴定神经内分泌GnRH神经元。 我们 将进行ICC染色DA，b-E，或GABA神经元与胶体 金，以确定哪些传入介导E2反馈。 类型和数量 将通过EM评估突触的数量并进行量化。 使用相同的 方案，我们将对P4受体进行ICC染色，并确定 哪些传入神经介导P4反馈。 我们将重复这些研究， 早期卵泡和中期黄体期动物检查反馈部位 以及改变类固醇水平的影响。 灵长类动物的青春期包括重新激活处于休眠状态的青春期前 GnRH脉冲发生器。 虽然这意味着有能力的GnRH神经元 系统存在于青少年，神经解剖学变化， 增强脉冲式GnRH分泌还有待评估。(3)我们将 确定GnRH神经元连接或传入突触是否改变 在青春期前后。 我们将进行双重EM ICC染色 GnRH的。神经元（DAB）及其传入（胶体金）， 青春期前和成年雄性猴子。 我们将量化和比较 GnRH神经元相互作用的类型、特征和数量， 传入突触 使用系列血浆LH测量（RIA），我们将 鉴定青春期（夜间LH峰）动物，进行ICC染色， 并评估所观察到的神经变化的参与。 在随后 研究中，我们将检查青春期前和青春期前后的雌性猴子， 量化它们的GnRH神经相互作用。 我们将把数据与 先前在成年骑自行车的女性中获得，并将观察到的 与青春期事件相关的变化。 这些研究一起 将阐明GnRH神经元控制的性质， 脉冲式GnRH分泌，并评估神经解剖学变化 到青春期的开始