红耳龟(Trachemys scripta elegans)中胃饥饿素的结构测定及组织化学定位

Structural determination and histochemical localization of ghrelin in the red-eared slider turtle, Trachemys scripta elegans.

作者信息

Kaiya Hiroyuki, Sakata Ichiro, Kojima Masayasu, Hosoda Hiroshi, Sakai Takafumi, Kangawa Kenji

机构信息

Department of Biochemistry, National Cardiovascular Center Research Institute, 5-7-1 Fujishirodai, Suita, Osaka 565-8565, Japan.

出版信息

Gen Comp Endocrinol. 2004 Aug;138(1):50-7. doi: 10.1016/j.ygcen.2004.05.005.

Abstract

We purified ghrelin peptide and determined the cDNA sequence encoding the precursor protein from the stomach of the red-eared slider turtle, Trachemys scripta elegans. The Trachemys ghrelin is comprised of 25-amino acids and has the sequence GSSFLSPEYQNTQQRKDPKKHTKLN. The third serine residue was modified by n-octanoic (C8:0), decanoic (C10:0) or unsaturated decanoic acid (C10:1). The carboxyl-terminal end of the peptide was not amidated, as seen in the ghrelins of other land vertebrates. Quantitative real-time PCR analysis revealed high levels of gene expression in the stomach and moderate levels in the large intestine and pancreas. Histochemical studies of turtle stomach revealed that ghrelin-immunopositive (ghrelin-ip) cells, which were small and round, were observed in the mucosal layer of the stomach but not in the myenteric plexus, and ghrelin-mRNA-expressing (ghrelin-ex) cells detected by in situ hybridization were scattered in a similar distribution as ghrelin-ip cells. These results indicate that ghrelin is present in reptiles.

摘要

我们从红耳龟(Trachemys scripta elegans)的胃中纯化了胃饥饿素肽,并确定了编码前体蛋白的cDNA序列。红耳龟的胃饥饿素由25个氨基酸组成,序列为GSSFLSPEYQNTQQRKDPKKHTKLN。第三个丝氨酸残基被正辛酸(C8:0)、癸酸(C10:0)或不饱和癸酸(C10:1)修饰。该肽的羧基末端没有酰胺化,这与其他陆地脊椎动物的胃饥饿素情况相同。定量实时PCR分析显示,该基因在胃中表达水平高,在大肠和胰腺中表达水平中等。对龟胃的组织化学研究表明,胃饥饿素免疫阳性(ghrelin-ip)细胞小而圆,在胃黏膜层中可见,但在肌间神经丛中未见,通过原位杂交检测到的胃饥饿素mRNA表达(ghrelin-ex)细胞分布与ghrelin-ip细胞相似。这些结果表明胃饥饿素存在于爬行动物中。

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