Signal Transduction Mechanisms in Neuroendocrine Neurons

描述（由申请人提供）：下丘脑室旁核（PVH）的含促肾上腺皮质激素释放激素（CRH）的神经元对于应激的适应性反应至关重要，其破坏可能与应激相关的病理学有关。本训练计划的长期目标是开始建立引起应激反应的刺激、将该信息传递给PVH的神经通路以及PVH神经元对该信息的协调反应之间的精确关系。具体的目标都围绕着如何后脑起源的儿茶酚胺（CA）传入帮助触发基因表达的CRH神经内分泌神经元的啮齿动物PVH的代谢应激，2-脱氧葡萄糖（2-DG）。建议的培训包括：（1）免疫组织化学和原位杂交，以跟踪CRH神经元活动的细胞标志物（例如，MAP激酶、CREB、crh hnRNA），（2）静脉内注射逆行免疫毒素2-DG或PVH以分别选择性刺激或损伤CA传入系统，和（3）PVH显微注射受体拮抗剂或酶抑制剂以探测CRH神经元的信号传导机制。这次培训将进一步推动我的职业目标，即使用体内方法研究细胞信号。

DESCRIPTION (provided by applicant): Corticotropin-releasing hormone (CRH)-containing neurons of the hypothalamic paraventricular nucleus (PVH) are critical for adaptive responses to stress, disruptions of which may be linked to stress-related pathologies. The long-term objective of this training proposal is to begin establishing the precise relationships among stimuli that evoke stress responses, the neural pathways conveying that information to the PVH, and the coordinated response of PVH neurons to such information. Specific aims all center around how the hindbrain-originating catecholaminergic (CA) afferents help trigger gene expression in CRH neuroendocrine neurons of the rodent PVH in response to the metabolic stressor, 2-deoxyglucose (2-DG). Proposed training includes: (1) immunohistochemistry and in situ hybridization to track cellular markers of CRH neuronal activity (e.g., MAP kinase, CREB, crh hnRNA), (2) intravenous 2-DG or PVH injection of a retrograde immunotoxin to selectively stimulate or lesion, respectively, the CA afferent system, and (3) PVH microinjections of receptor antagonists or enzyme inhibitors to probe the signaling machinery of CRH neurons. This training will further my career goal of studying cellular signaling using an in vivo approach.