Identification, localization and functional Implication of 26RFa ortholog peptide in the brain of zebra finch (Taeniopygia guttata)
Identification, localization and functional Implication of 26RFa ortholog peptide in the brain of zebra finch (Taeniopygia guttata)
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斑胸草雀 (Taeniopygia guttata) 大脑中 26RFa 直系同源肽的鉴定、定位和功能意义
DOI：
10.1111/j.1365-2826.2011.02179.x
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发表时间：
2011
期刊：
影响因子：
--
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et al
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作者：
Y.Tobari;K.Tsutsui;et al
在多种脊椎动物的下丘脑中发现了几种含有Arg-Phe-NH2(RFA)序列的神经肽。本研究旨在从斑雀脑中分离出新的RFA多肽。用一种识别禽类RFA多肽的抗体，通过免疫亲和纯化来分离多肽。分离的多肽由25个氨基酸组成，C-末端有RFA。序列为SGTLGNLAEEINGYNRRKGGFTFRFa。与脊椎动物26RFa的比对表明，所鉴定的多肽为斑雀26RFa。我们还克隆了编码该多肽的前体基因。对该基因的同源性分析表明，该基因在脊椎动物中高度保守。此外，我们还在斑雀脑中克隆了编码26RFa受体G蛋白偶联受体103(GPR103)的基因。GPR103基因编码一个432个氨基酸的蛋白质，有7个跨膜区。在脑中的原位杂交分析表明，26RFa mRNA的表达仅限于下丘脑前内侧区、下丘脑腹内侧核和下丘脑外侧区，这三个区域参与了摄食行为的调节，而GPR103 mRNA除分布在下丘脑核外，还分布在大脑各区。在自由喂养的雄性斑马雀体内集中给药时，26RFa在注射后24 h增加了摄食量，而体重没有变化。禁食10 h间脑GPR103mRNA表达上调，提示下丘脑26RFa-ITS受体系统在斑雀摄食量和能量平衡的中枢调控中起重要作用。
Several neuropeptides with the C‐terminal Arg‐Phe‐NH2(RFa) sequence have been identified in the hypothalamus of a variety of vertebrates. The present study was conducted to isolate novel RFa peptides from the zebra finch brain. Peptides were isolated by immunoaffinity purification using an antibody that recognises avian RFa peptides. The isolated peptide consisted of 25 amino acids with RFa at its C‐terminus. The sequence was SGTLGNLAEEINGYNRRKGGFTFRFa. Alignment of the peptide with vertebrate 26RFa has revealed that the identified peptide is the zebra finch 26RFa. We also cloned the precursor cDNA encoding this peptide. Synteny analysis of the gene showed a high conservation of this gene among vertebrates. In addition, we cloned the cDNA encoding a putative 26RFa receptor, G protein‐coupled receptor 103 (GPR103) in the zebra finch brain. GPR103 cDNA encoded a 432 amino acid protein that has seven transmembrane domains.In situhybridisation analysis in the brain showed that the expression of 26RFa mRNA is confined to the anterior‐medial hypothalamic area, ventromedial nucleus of the hypothalamus and the lateral hypothalamic area, the brain regions that are involved in the regulation of feeding behaviour, whereas GPR103 mRNA is distributed throughout the brain in addition to the hypothalamic nuclei. When administered centrally in free‐feeding male zebra finches, 26RFa increased food intake 24 h after injection without body mass change. Diencephalic GPR103 mRNA expression was up‐regulated by fasting for 10 h. Our data suggest that the hypothalamic 26RFa‐its receptor system plays an important role in the central control of food intake and energy homeostasis in the zebra finch.