Pfizer Global Research and Development, Autism Research Unit, Eastern Point Road, Groton, CT 06340, USA.
Neuroscience. 2011 Jan 13;172:153-63. doi: 10.1016/j.neuroscience.2010.10.020. Epub 2010 Oct 13.
The neuropeptide S receptor (NPSR) is a G-protein coupled receptor that is potently activated by the linear 20 amino acid peptide, neuropeptide S (NPS). Central administration of NPS promotes arousal and anxiolytic-like effects in rodents, and fails to promote such effects in NPSR knockout animals or in the presence of NPSR-selective antagonists. In situ hybridization (ISH) studies in rat brain have revealed that the mRNAs encoding the NPS precursor and the NPS receptor are expressed at high levels in discrete regions of the rat CNS. The distribution of the NPSR protein in brain has not been reported due to a lack of available antibodies. We have generated and validated a NPSR-specific antibody and used it to determine the distribution of the NPSR in male Sprague-Dawley (SD) rat brain. The anti-NPSR antibody identified a single protein by Western blot with an estimated molecular weight of 65 kD, which was prevented by pre-incubation of the antibody with the immunizing peptide. The protein distribution identified with this antibody in rat brain was consistent both with the mRNA distribution identified by in situ hybridization, and to the localization pattern identified by a second NPSR-specific antibody against a distinct NPSR epitope. NPSR protein was identified in the medial amygdala (MeA), substantia nigra pars compacta, subiculum, dorsal raphe, and several hypothalamic and thalamic regions. Additionally, NPSR protein was localized in the pyramidal cell layer of the ventral hippocampus, the medial habenula (MHb), and was widely distributed in the cortex. The distribution of NPSR protein provides further insight into the organization of the NPS system and may guide future studies on the role of the NPSR in brain.
神经肽 S 受体（NPSR）是一种 G 蛋白偶联受体，能被线性 20 个氨基酸肽神经肽 S（NPS）强烈激活。NPS 的中枢给药能促进啮齿动物的觉醒和抗焦虑样效应，而在 NPSR 敲除动物或存在 NPSR 选择性拮抗剂时，NPS 则不能促进此类效应。大鼠脑内原位杂交（ISH）研究显示，编码 NPS 前体和 NPS 受体的 mRNAs 在大鼠中枢神经系统的离散区域高水平表达。由于缺乏可用的抗体，NPSR 蛋白在脑中的分布尚未报道。我们已经产生并验证了一种 NPSR 特异性抗体，并使用它来确定雄性 Sprague-Dawley（SD）大鼠脑中 NPSR 的分布。该抗 NPSR 抗体通过 Western blot 鉴定出一种估计分子量为 65 kD 的单一蛋白，该蛋白可通过将抗体与免疫肽预孵育来阻止。该抗体在大鼠脑中鉴定出的蛋白分布与原位杂交鉴定出的 mRNA 分布一致，也与针对不同 NPSR 表位的第二种 NPSR 特异性抗体确定的定位模式一致。NPSR 蛋白在中杏仁核（MeA）、黑质致密部、下托、背侧中缝核和几个下丘脑和丘脑区域被鉴定出。此外，NPSR 蛋白定位于腹侧海马的锥体细胞层、中脑缰核（MHb），并广泛分布于皮质。NPSR 蛋白的分布为 NPS 系统的组织提供了进一步的深入了解，并可能指导未来关于 NPSR 在大脑中的作用的研究。
