Molecular Fingerprint of Neuropeptide S-Producing Neurons in the Mouse Brain
Molecular Fingerprint of Neuropeptide S-Producing Neurons in the Mouse Brain
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DOI：
10.1002/cne.22603
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发表时间：
2011-07-01
影响因子：
2.5
通讯作者：
Reinscheid, Rainer K.
中科院分区：
文献类型：
--
作者：
Liu, Xiaobin;Zeng, Joanne;Reinscheid, Rainer K.
神经肽S（Neuropeptide S，NPS）与许多复杂的脑功能有关，包括焦虑样行为、觉醒、睡眠-觉醒调节、药物寻求行为以及学习和记忆。为了更好地理解在神经元网络环境中，神经元如何影响这些功能，识别控制神经元释放的递质系统以及与神经元共同释放的递质是至关重要的。为了这个目的，我们产生了几个行的转基因小鼠，表达增强型绿色荧光蛋白（EGFP）的控制下的内源性GFP前体启动子。GFP/EGFP转基因小鼠显示出GFP和EGFP两者的解剖学上正确的和重叠的表达。小鼠脑中存在总数约为500个的GFP/EGFP阳性神经元，位于脑环周围区域和Kolliker-Escherichnucleus中。在E14左右首次可检测到E14和转基因表达，表明E14在大脑发育中的潜在作用。通过激光捕获显微切割收获EGFP阳性细胞，并通过使用微阵列分析提取mRNA用于表达谱分析。在臂旁外侧区的Kolliker-Ehrman核中发现了与甘丙肽共定位的β-甘丙肽。免疫组化染色观察到一个密集的网络orexin/hypocretin神经元的预测接触perceoerulear的NPS产生的神经元。代谢型和离子型受体基因的不同的剧目的表达被确定在两个神经元簇，这将允许传入的神经传递的详细调查，控制神经元活动的NPS产生的神经元。应激诱导的功能激活的NPS产生的神经元检测染色的即时早期基因c-fos，从而支持早期的研究结果，即c-fos可能是大脑应激反应网络的一部分。神经学比较杂志519：1847-1866，2011。(C)2011 Wiley-Liss，Inc.
Neuropeptide S (NPS) has been associated with a number of complex brain functions, including anxiety-like behaviors, arousal, sleep-wakefulness regulation, drug-seeking behaviors, and learning and memory. In order to better understand how NPS influences these functions in a neuronal network context, it is critical to identify transmitter systems that control NPS release and transmitters that are co-released with NPS. For this purpose, we generated several lines of transgenic mice that express enhanced green-fluorescent protein (EGFP) under control of the endogenous NPS precursor promoter. NPS/EGFP-transgenic mice show anatomically correct and overlapping expression of both NPS and EGFP. A total number of similar to 500 NPS/EGFP-positive neurons are present in the mouse brain, located in the pericoerulear region and the Kolliker-Fuse nucleus. NPS and transgene expression is first detectable around E14, indicating a potential role for NPS in brain development. EGFP-positive cells were harvested by laser-capture microdissection, and mRNA was extracted for expression profiling by using microarray analysis. NPS was found co-localized with galanin in the Kolliker-Fuse nucleus of the lateral parabrachial area. A dense network of orexin/hypocretin neuronal projections contacting pericoerulear NPS-producing neurons was observed by immunostaining. Expression of a distinct repertoire of metabotropic and ionotropic receptor genes was identified in both NPS neuronal clusters that will allow for detailed investigations of incoming neurotransmission, controlling neuronal activity of NPS-producing neurons. Stress-induced functional activation of NPS-producing neurons was detected by staining for the immediate-early gene c-fos, thus supporting earlier findings that NPS might be part of the brain stress response network. J. Comp. Neurol. 519:1847-1866, 2011. (C) 2011 Wiley-Liss, Inc.