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characterization of the novel duplicated PRLR gene at the late-feathering K locus in Lohmann chickens.

Characterization of the novel duplicated PRLR gene at the late-feathering K locus in Lohmann chickens.

机构信息

Key Laboratory of Bio-resources and Eco-environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu 610065, People's Republic of China.

出版信息

J Mol Endocrinol. 2013 Sep 13;51(2):261-76. doi: 10.1530/JME-13-0068. Print 2013 Oct.

Abstract

A partial duplication of the prolactin (PRL) receptor gene (designated as dPRLR) has been identified at the late-feathering (LF) K locus on chromosome Z of some chicken strains recently, implying that dPRLR is probably a candidate gene associated with LF development in chickens. However, little is known about the structure, functionality, and spatiotemporal expression of the dPRLR gene in chickens. In this study, using 3'-RACE and RT-PCR, the full-length cDNA of the dPRLR obtained from the kidneys of male Lohmann layer chickens carrying a K allele was cloned. The cloned dPRLR is predicted to encode a membrane-spanning receptor of 683 amino acids, which is nearly identical to the original PRLR, except for its lack of a 149-amino acid C-terminal tail. Using a 5× STAT5-Luciferase reporter system and western blot analysis, we demonstrated that dPRLR expressed in HepG2 cells could be potently activated by chicken PRL and functionally coupled to the intracellular STAT5 signaling pathway, suggesting that dPRLR may function as a novel receptor for PRL. RT-PCR assays revealed that similar to the original PRLR gene, dPRLR mRNA is widely expressed in all embryonic and adult tissues examined including the skin of male Lohmann chickens with a K allele. These findings, together with the expression of PRL mRNA detected in the skin of embryos at embryonic day 20 and 1-week-old chicks, suggest that skin-expressed dPRLR and PRLR, together with plasma and skin-derived PRL, may be involved in the control of the LF development of chicks at hatching. Moreover, the wide tissue expression of dPRLR implies that dPRLR may regulate other physiological processes of chickens carrying the K allele.

摘要

最近,在一些鸡品系的 Z 染色体晚期羽毛(LF)K 基因座上发现了催乳素(PRL)受体基因(命名为 dPRLR)的部分重复,这表明 dPRLR 可能是与鸡 LF 发育相关的候选基因。然而,关于鸡 dPRLR 基因的结构、功能和时空表达知之甚少。在这项研究中,使用 3'-RACE 和 RT-PCR,从携带 K 等位基因的雄性洛曼层鸡肾脏中克隆得到 dPRLR 的全长 cDNA。克隆的 dPRLR 预测编码一个跨膜受体,由 683 个氨基酸组成,与原始 PRLR 几乎相同,只是缺少 149 个氨基酸的 C 端尾部。使用 5×STAT5-Luciferase 报告系统和 Western blot 分析,我们证明了在 HepG2 细胞中表达的 dPRLR 可以被鸡 PRL 强烈激活,并与细胞内 STAT5 信号通路功能偶联,表明 dPRLR 可能作为 PRL 的新型受体发挥作用。RT-PCR 检测表明,与原始 PRLR 基因相似,dPRLR mRNA 在所有胚胎和成年组织中广泛表达,包括携带 K 等位基因的雄性洛曼鸡的皮肤。这些发现,以及在胚胎 20 天和 1 周龄雏鸡皮肤中检测到的 PRL mRNA 的表达,表明皮肤表达的 dPRLR 和 PRLR 与血浆和皮肤来源的 PRL 一起,可能参与控制孵化时小鸡的 LF 发育。此外,dPRLR 的广泛组织表达表明,dPRLR 可能调节携带 K 等位基因的鸡的其他生理过程。

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