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鉴定猪 G 蛋白偶联受体 41 和 43 基因及其在不同组织和发育阶段的表达模式。

Identification of the porcine G protein-coupled receptor 41 and 43 genes and their expression pattern in different tissues and development stages.

机构信息

Laboratory of Gastrointestinal Microbiology, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, Jiangsu, China.

Laboratory of Gastrointestinal Microbiology, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, Jiangsu, China; Key Lab of Animal Physiology and Biochemistry, Ministry of Agriculture, Nanjing, Jiangsu, China.

出版信息

PLoS One. 2014 May 19;9(5):e97342. doi: 10.1371/journal.pone.0097342. eCollection 2014.

Abstract

Short-chain fatty acids (SCFAs) are not only an important energy source, but they also play a regulatory role in various physiological processes in humans and rodents. Current studies, mostly in humans and rodents, have revealed that SCFAs acted as endogenous ligands for G protein-coupled receptor GPR41 and GPR43. Whether proteins similar to human GPR41 and GPR43 mediate the regulatory effects of SCFAs in swine remains unclear to date. The aims of this study were to determine whether GPR41 and GPR43 genes are expressed in porcine different tissues; and whether the expression of GPR41 and GPR43 is tissue-specific and/or time-associated. The alignment results showed that pig chromosome 6 contained GPR41 and GPR43 genes. Reverse transcription polymerase chain reaction (RT-PCR) indicated that GPR41 and GPR43 were expressed in porcine various tissues. The 2218 bp and 1908 bp nucleotide sequence representing the full-length cDNA sequence of porcine GPR41 and GPR43 was obtained from the ileum and spleen using rapid amplification of cDNA ends (RACE), which were capable of encoding 335 and 329 amino acid sequences, respectively. The structure prediction revealed that porcine GPR41 and GPR43 proteins had seven putative trans-membrane domains. The real-time PCR results indicated that GPR41 and GPR43 were expressed throughout the developmental stages in a tissue-specific and time-associated manner. GPR41 and GPR43 were most highly expressed in the ileum (P<0.01) and the spleen (P<0.01), respectively. Western blot results showed that porcine GPR41 and GPR43 proteins were expressed in a variety of porcine tissues, including the spleen, ileum, colon, and adipose tissue. In situ GPR41 and GPR43 immunoreactivities were observed through immunohistochemistry in the spleen, ileum, colon, and adipose tissue. In conclusion, the pig genome encoded GPR41 and GPR43 genes, and these two genes were detected in a variety of porcine tissues and expressed in tissue-specific and time-associated manner.

摘要

短链脂肪酸(SCFAs)不仅是重要的能量来源,而且在人和啮齿动物的各种生理过程中发挥调节作用。目前的研究主要在人和啮齿动物中表明,SCFAs 作为 G 蛋白偶联受体 GPR41 和 GPR43 的内源性配体发挥作用。目前尚不清楚是否类似于人类 GPR41 和 GPR43 的蛋白质介导 SCFAs 在猪中的调节作用。本研究的目的是确定 GPR41 和 GPR43 基因是否在猪的不同组织中表达;以及 GPR41 和 GPR43 的表达是否具有组织特异性和/或时间相关性。序列比对结果表明,猪 6 号染色体包含 GPR41 和 GPR43 基因。逆转录聚合酶链反应(RT-PCR)表明 GPR41 和 GPR43 在猪的各种组织中表达。使用快速扩增 cDNA 末端(RACE)从回肠和脾脏中获得代表全长 cDNA 序列的 2218 bp 和 1908 bp 核苷酸序列,它们分别能够编码 335 和 329 个氨基酸序列。结构预测表明,猪 GPR41 和 GPR43 蛋白具有 7 个潜在的跨膜结构域。实时 PCR 结果表明,GPR41 和 GPR43 在组织特异性和时间相关性方面在整个发育阶段表达。GPR41 和 GPR43 在回肠(P<0.01)和脾脏(P<0.01)中表达最高。Western blot 结果表明,猪 GPR41 和 GPR43 蛋白在多种猪组织中表达,包括脾脏、回肠、结肠和脂肪组织。通过免疫组织化学观察到猪 GPR41 和 GPR43 免疫反应性在脾脏、回肠、结肠和脂肪组织中存在。总之,猪基因组编码 GPR41 和 GPR43 基因,并且在多种猪组织中检测到这两个基因,并以组织特异性和时间相关性的方式表达。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2b97/4026140/d72e0e4ab1f1/pone.0097342.g001.jpg

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