绵羊下丘脑Ghrlein通过神经元投射参与繁殖调控的研究

胃肠道产生的肽类激素Ghrelin可通过雌二醇受体(ERα)激活下丘脑内NPY/AgRP神经元促进采食，已知能量代谢平衡是生殖系统的重要物质基础，但绵羊下丘脑Ghrelin神经元通过能量代谢平衡参与繁殖调控的作用方式尚有诸多问题有待澄清。为此本研究首先通过PRV病毒逆行追踪确定下丘脑与卵巢间是否存在与Ghrelin相关的神经作用通路，免疫荧光三标检测发情前、中、后Ghrelin/AgRP/ERα和NPY/GHSR/ERα在下丘脑内的神经投射差异，分析血液内Ghrelin和E2间的作用关系，并将下丘脑神经元体外培养通过正调和负调检测第二信使Ca2+和cAMP的变化，以及ERα/UCP1/pERK-MAPK/pAMPK/pAKT/CaMKII分子和蛋白水平的变化，以期从神经通路、神经元间投射、电生理和信号通路的角度，解释Ghrlein在“能量代谢-神经内分泌”联动中的作用及与繁殖间的关系。

Ghrelin from gastrointestinal peptide hormone induces food intake and is associated with increased NPY/AgRP expression by ERα in the hypothalamus. It is well known fact that reproductive function is highly dependent on food intake, body status or metabolic disorders. However, it remains unclear the relationship of neurons of ghrelin involve the reproduction by metabolism balance within sheep hypothalamus. we therefore investigate connection with hypothalamus and ovary using pseudorabies virus (PRV) for the retrograde transsynaptic. Then confirm co-expression neurons of Ghrelin/AgRP/ERα and NPY/GHSR/ERα by triple fluorescence immunohistochemistry at different estrus cycle, also plasma concentrations of Ghrelin and E2 were compared. Based on above evidences we next will evaluate the level of Ca2+ and cAMP in different treatment after culturing the neurons from hypothalamus, and ERα/UCP1/pERK-MAPK/pAMPK/pAKT/CaMKII were analyzed in vitro and vivo to understand the how the ghrelin neurons within hypothalamus involve the regulation of never pathway. Then make sure the distribution net and potential roles of ghrelin in ovine reproduction and provided some theories for the hypothesis that the relationship between energy metabolism and neuroendocrinology mediate the reproduction.