NEURAL INFLUENCE ON HYPOPHYSEAL-GONADAL FUNCTION

不孕症通常是由于月经周期中断和/或 排卵：已知由特定下丘脑驱动的过程 信号。通过监测自然排卵或 去卵巢(OVX)猕猴，我们观察到 促性腺激素释放激素(GnRH)分泌 17β(E_2)诱导黄体生成素(L H)升高。不少于14 神经递质/神经肽(S)参与促性腺激素释放激素的调节 分泌物。因为在促性腺激素释放激素神经元上发现了肾上腺素能受体，我们 最初关注去甲肾上腺素(NE)和神经肽Y(NPY)，它们 在去甲肾上腺素能细胞中共定位并刺激GnRH基因表达 以及包括猕猴在内的受E2调节的动物的分泌物。目标1a意志 排卵前卵巢E_2升高是否刺激去甲肾上腺素和神经肽Y 分泌与雌激素诱导的促性腺激素释放激素/促黄体生成素激增有关。目标1b将 选择性阻断去甲肾上腺素或神经肽Y受体活性 拮抗剂可阻断促性腺激素释放激素/促黄体素释放激素峰。连续推拉灌流或 微透析将在下丘脑的内侧基底(MBH)进行。 使用或不使用哌唑嗪(NE)治疗的完整猴子和去卵巢猴子 拮抗剂)和(Ac[3-(2，6二氯苯基)酪氨酸(27，36)D-苏氨酸(32)]NPY-(27- 36)酰胺(NPY拮抗剂)。灌流液中去甲肾上腺素、神经肽Y和 GnRH将通过高效液相色谱和 放射免疫分析。目标2解决了形态测量和 E2如何影响NPY/NE/GnRH分泌的细胞内证据。目标 2A将确定雌激素受体是否定位于NE和/或NPY 神经元，如果在NE神经元产生NPY，如果NE/NPY终末 与下丘脑GnRH神经元突触。人脑出血的组织学切片 和脑干(即蓝斑/外侧被盖)将被检查 对于表达雌激素受体mRNAs的神经元的定位(通过 原位杂交)和去甲肾上腺素(即多巴胺-β-羟基酶)、神经肽Y和 GnRH(免疫细胞化学法)。Aim 2b将量化雌二醇诱导的变化 在NPY和NE(即酪氨酸羟基酶)中确定了mRNA的含量 核糖核酸酶保护法测定脑干和下丘脑的基因座。 AIMS 3和4研究突触NE分泌如何与 可能的神经调节剂来调节GnRH的释放。目标3a将决定 如果阻断去甲肾上腺素转运体活性(突触前去甲肾上腺素再摄取) 抗抑郁药地昔帕明强烈刺激去甲肾上腺素，从而释放促性腺激素释放激素。 目标3b将显示持续输注地塞帕明抑制NE/GnRH 分泌，并确定GnRH脉冲是否可以通过以下脉冲重新启动 NE或神经兴奋性氨基酸(即N-甲基-D、L-天冬氨酸)。 将使用植入去卵巢猕猴MBH的微透析套管 对于急性(3a)或持续(3b)地塞帕明治疗，脉冲式NE或 N-甲基-D，L-天冬氨酸输注(3b)和微透析液收集 NE和GnRH测量(3a，b)。Aim 4a将通过现场检查 杂交法：慢性下丘脑内注射地西帕明是否能抑制 退行性NE转运体、酪氨酸羟化酶或NPY mRNAs在 脑干NE细胞。Aim 4b将寻找E2下调的证据 去甲肾上腺素转运体信息在去甲肾上腺素神经元。这些研究将表明 特定神经化学物质对促性腺激素释放激素分泌的重要性 对生育和生殖疾病的生物学和病理学知识 神经抑郁，不育的雌性灵长类动物。

Infertility often results from disruption of menstrual cyclicity and/or ovulation: processes known to be driven by specific hypothalamic signals. By monitoring neurosecretions in either naturally ovulating or ovariectomized (OVX) rhesus macaques, we observed a 10-fold increase in gonadotropin-releasing hormone (GnRH) secretion during an estradiol- 17beta (E2)-induced luteinizing hormone (LH) surge. No less than 14 neurotransmitter/neuropeptide(s) are implicated in the regulation of GnRH secretion. Because adrenergic receptors are found on GnRH neurons, we initially focus on norepinephrine (NE) and neuropeptide Y (NPY), which are colocalized in noradrenergic cells and stimulate GnRH gene expression and secretion in E2-conditioned animals, including macaques. Aim 1a will resolve if the preovulatory rise in ovarian E2 stimulates NE and NPY secretion in conjunction with the E2-induced GnRH/LH surge. Aim 1b will decipher if blockade of either NE or NPY receptor activity by selective antagonists blocks the GnRH/LH surge. Continuous push-pull perfusion or microdialysis will be performed in the mediobasal hypothalamus (MBH) of both intact and OVX monkeys with or without treatment of prazosin (NE antagonist) and (Ac[3-(2,6 dichlorobenzyl)tyr(27,36)D-Thr(32)]NPY-(27- 36)amide (NPY antagonist). Changes in perfusate levels of NE, NPY and GnRH will be analyzed by high-performance liquid chromatography and radioimmunoassays, respectively. Aim 2 addresses the morphometric and intracellular evidence of how E2 influences NPY/NE/GnRH secretion. Aim 2a will determine if estrogen receptors are localized in NE and/or NPY neurons, if NPY is produced in NE neurons, and if NE/NPY terminals synapse with hypothalamic GnRH neurons. Histological sections of the MBH and brainstem (i.e., locus coeruleus/lateral tegmentum) will be examined for the localization of neurons that express estrogen receptor mRNAs (by in situ hybridization) and NE (i.e., dopamine-beta-hydroxylase), NPY and GnRH (by immunocytochemistry). Aim 2b will quantitate E2-induced changes in NPY and NE (i.e., tyrosine hydroxylase) mRNA contents in identified loci of brainstem and hypothalamus by ribonuclease protection assay. Aims 3 and 4 examine how synaptic NE secretion interacts with other putative neuromodulators to regulate GnRH release. Aim 3a will determine if blockade of NE transporter activity (presynaptic NE reuptake) by the antidepressant, desipramine, acutely stimulates NE, hence GnRH, release. Aim 3b will show that continuous desipramine infusion suppresses NE/GnRH secretion, and establish if GnRH pulses can be reinitiated by pulses of NE or neuroexcitatory amino acids (i.e., N-methyl-D,L-aspartate). Microdialysis cannulae implanted in the MBH of OVX monkeys will be used for acute (3a) or continuous (3b) desipramine treatment, pulsatile NE or N-methyl-D,L-aspartate infusion (3b) and microdialysate collection for NE and GnRH measurements (3a,b). Aim 4a will examine by in situ hybridization whether chronic intrahypothalamic desipramine can suppress retrogradely NE transporter, tyrosine hydroxylase, or NPY mRNAs in brainstem NE cells. Aim 4b will search for evidence of E2 downregulation of NE transporter message in NE neurons. These studies will indicate the importance of specific neurochemical on GnRH secretion thereby increasing the knowledge of biological and pathological events in fertile and neurally depressed, infertile female primates.