Characterization of corticotropin-releasing hormone neurons in the paraventricular nucleus of the hypothalamus of Crh-IRES-Cre mutant mice.

Characterization of corticotropin-releasing hormone neurons in the paraventricular nucleus of the hypothalamus of Crh-IRES-Cre mutant mice.
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DOI:
10.1371/journal.pone.0064943
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发表时间:
2013
期刊:
影响因子:
3.7
通讯作者:
Bains JS
Bains JS
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Wamsteeker Cusulin JI;Füzesi T;Watts AG;Bains JS

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下丘脑室旁核(PVN)中的促肾上腺皮质激素释放激素(CRH)神经元启动和控制神经内分泌反应,以应对心理和生理应激。然而,由于缺乏充分靶向或可视化这一细胞群体的工具,对CRH神经元的生理学研究一直受到阻碍。在这里,我们表征了表达td番茄荧光团的小鼠PVN中的CRH神经元,td番茄荧光团是由最近开发的CRH-IRES-Cre驱动程序和Ai14 Cre报告基因小鼠品系杂交产生的。在CRH-IRES-Cre中含有PVN神经元的td番茄;Ai14小鼠在没有二次检测方法的情况下很容易被观察到。这些神经元以神经内分泌为主,大量表达CRH蛋白,但不表达其他PVN表型神经肽。在急性应激后,绝大多数td番茄细胞表达神经元激活标记c-Fos。最后,tdTomato PVN神经元表现出均一的内在生物物理和突触特性,并可以通过病毒Cre驱动的通道视紫红质表达来进行光遗传操作。这些观察突出了突变小鼠CRH神经元的基本细胞类型特征,为其未来在探索内分泌应激反应的神经生理学中的应用提供了验证。
Corticotropin-releasing hormone (CRH)-containing neurons in the paraventricular nucleus of the hypothalamus (PVN) initiate and control neuroendocrine responses to psychogenic and physical stress. Investigations into the physiology of CRH neurons, however, have been hampered by the lack of tools for adequately targeting or visualizing this cell population. Here we characterize CRH neurons in the PVN of mice that express tdTomato fluorophore, generated by crosses of recently developed Crh-IRES-Cre driver and Ai14 Cre-reporter mouse strains. tdTomato containing PVN neurons in Crh-IRES-Cre;Ai14 mice are readily visualized without secondary-detection methods. These neurons are predominantly neuroendocrine and abundantly express CRH protein, but not other PVN phenotypic neuropeptides. After an acute stress, a large majority of tdTomato cells express neuronal activation marker c-Fos. Finally, tdTomato PVN neurons exhibit homogenous intrinsic biophysical and synaptic properties, and can be optogenetically manipulated by viral Cre-driven expression of channelrhodopsin. These observations highlight basic cell-type characteristics of CRH neurons in a mutant mouse, providing validation for its future use in probing neurophysiology of endocrine stress responses.
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