Morin S Michelle, Gehlert Donald R
Neuroscience, Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN 46285, USA.
J Mol Neurosci. 2006;29(2):109-14. doi: 10.1385/JMN:29:2:109.
The pharmacology and brain mRNA distribution of the neuropeptide Y (NPY) rat Y5 (rY5) receptor has led to the hypothesis that this receptor might mediate the hypothalamic feeding response to NPY in addition to many other physiologic functions. However, through the use of autoradiographic techniques, only very low levels of Y5-like immunoreactive (Y5-ir) binding are detected in the rat brain. To localize the Y5 protein in the rat brain, polyclonal antibodies were raised to the carboxyl terminus of the rY5 receptor. The resulting antisera were affinity purified and characterized by specific binding to HEK293 cells that had been stably transfected with the rY5 receptor. Utilizing immunohistochemical techniques, we found a discrete pattern of Y5-ir in the rat brain. In initial studies, very low levels of Y5-ir were detected, and TSA amplification was required to visualize the staining. Areas with the highest levels of expression include the piriform cortex, supraoptic nucleus, and hippocampus. Areas with moderate levels of expression include the lateral septum, amygdala, arcuate nucleus, paraventricular hypothalamic nucleus, locus coeruleus, and cerebellum. With several exceptions, this pattern of distribution is consistent with earlier reports of rY5 mRNA and receptor protein expression.
神经肽Y(NPY)大鼠Y5(rY5)受体的药理学及脑内mRNA分布情况引发了这样一种假说,即除了许多其他生理功能外,该受体可能介导下丘脑对NPY的进食反应。然而,通过放射自显影技术,在大鼠脑中仅检测到极低水平的Y5样免疫反应性(Y5-ir)结合。为了在大鼠脑中定位Y5蛋白,制备了针对rY5受体羧基末端的多克隆抗体。所得抗血清经亲和纯化,并通过与稳定转染了rY5受体的HEK293细胞的特异性结合进行表征。利用免疫组织化学技术,我们在大鼠脑中发现了Y5-ir的离散分布模式。在初步研究中,检测到极低水平的Y5-ir,需要TSA扩增才能使染色可视化。表达水平最高的区域包括梨状皮质、视上核和海马体。表达水平中等的区域包括外侧隔、杏仁核、弓状核、下丘脑室旁核、蓝斑和小脑。除了几个例外情况,这种分布模式与早期关于rY5 mRNA和受体蛋白表达的报道一致。