Signal Transduction Mechanisms in Neuroendocrine Neurons

描述(申请人提供)：下丘脑室旁核(PVH)中含有促肾上腺皮质激素释放激素(CRH)的神经元对于适应应激反应至关重要，这种反应的中断可能与应激相关的病理有关。该培训计划的长期目标是开始建立引起应激反应的刺激之间的精确关系，将该信息传递到PVH的神经通路，以及PVH神经元对此类信息的协调反应。特定的目标都集中在后脑起源的儿茶酚胺(CA)能传入如何帮助在啮齿动物PVH的CRH神经内分泌神经元中触发基因表达，以响应代谢应激源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追踪CRH神经元活性的细胞标记物(如MAP激酶、CREB、CRH hnRNA)；(2)静脉注射2-DG或PVH分别选择性刺激或损害CA传入系统；(3)PVH微量注射受体拮抗剂或酶抑制剂以探索CRH神经元的信号机制。这次培训将进一步推进我的职业目标，即使用活体方法研究细胞信号。

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.