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
复制标题
罗曼鸡晚羽 K 位点新型重复 PRLR 基因的表征。
DOI：
10.1530/jme-13-0068
复制
发表时间：
2013-10-01
影响因子：
3.5
通讯作者：
Wang, Yajun
中科院分区：
文献类型：
--
作者：
Bu, Guixian;Huang, Guian;Wang, Yajun
最近在一些鸡的Z染色体上的晚羽化（LF） K位点发现了部分重复的催乳素（PRL）受体基因（dPRLR），这表明dPRLR可能是鸡晚羽化（LF）发育相关的候选基因。然而，人们对鸡体内dprr基因的结构、功能和时空表达知之甚少。本研究利用3′-RACE和RT-PCR技术，克隆了携带K等位基因的雄性罗曼蛋鸡肾脏dPRLR的全长cDNA。预测克隆的PRLR编码683个氨基酸的跨膜受体，除了缺乏149个氨基酸的c端尾部外，与原始PRLR几乎相同。通过5 × STAT5-荧光素酶报告系统和western blot分析，我们证明了在HepG2细胞中表达的dPRLR可以被鸡PRL有效激活，并与细胞内STAT5信号通路功能偶联，这表明dPRLR可能是一种新的PRL受体。RT-PCR分析显示，与原PRLR基因相似，PRLR mRNA在所有胚胎和成体组织中广泛表达，包括携带K等位基因的雄性罗曼鸡的皮肤。上述结果，结合20日龄和1周龄雏鸡胚皮中PRL mRNA的表达，提示皮肤表达的dPRLR和PRLR以及血浆和皮肤源性PRL可能参与了雏鸡孵化时LF发育的控制。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。
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 x 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.