Department of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo, Tokyo 113-8657, Japan.
Gen Comp Endocrinol. 2010 May 15;167(1):27-34. doi: 10.1016/j.ygcen.2010.03.004. Epub 2010 Mar 11.
Prolactin (PRL)-releasing peptide (PrRP) has been reported as a strong candidate for a stimulating factor of both PRL secretion and expression in teleost species; however, there is no information available on its receptor. Here we report cDNA cloning and characterization of PrRP receptor expressed in the pituitary of Mozambique tilapia Oreochromis mossambicus. The deduced amino acid sequence of cDNA for tilapia PrRP receptor shared 50-83% homology with other vertebrate homologs. Intracellular calcium mobilization assay revealed that PrRP receptor responded to as low as 1nM order of tilapia PrRP, indicating its high affinity to PrRP. The expression of PrRP receptor was detected in the brain, pituitary, heart, spleen, kidney and rectum of freshwater (FW)- and seawater (SW)-adapted fish. There was no significant difference between FW and SW fish in transcription levels of PrRP receptor in the rostral pars distalis (RPD) of the pituitary. Similarly, the PrRP expression level in the whole brain was not changed by environmental salinity. Immunohistochemistry with a specific antibody showed that PrRP receptor was mainly localized in the cells of the RPD and neurohypophysis in the pituitary of both FW and SW tilapia. We also examined the effects of PrRP on PRL expression in primary-incubated PRL cells of FW tilapia; PrRP failed to stimulate PRL expression in PRL cells in vitro. These results suggest that in vivo stimulatory effects of PrRP on PRL gene expression reported in teleosts are presumably mediated by an unknown regulator secreted from the neurohypophysis expressing PrRP receptor.
催乳素(PRL)释放肽(PrRP)已被报道为刺激鱼类 PRL 分泌和表达的强有力候选因子;然而,目前尚无关于其受体的信息。在这里,我们报道了摩拉氏鲶 Oreochromis mossambicus 垂体中表达的 PrRP 受体的 cDNA 克隆和特性。cDNA 推导的鲶鱼 PrRP 受体的氨基酸序列与其他脊椎动物同源物具有 50-83%的同源性。细胞内钙动员测定表明,PrRP 受体对低至 1nM 浓度的鲶鱼 PrRP 有反应,表明其对 PrRP 具有高亲和力。PrRP 受体的表达在淡水(FW)和海水(SW)适应鱼的大脑、垂体、心脏、脾脏、肾脏和直肠中均有检测到。在垂体的前叶(RPD)中,FW 和 SW 鱼之间 PrRP 受体的转录水平没有显著差异。同样,环境盐度对全脑 PrRP 表达水平没有影响。用特异性抗体进行免疫组织化学染色显示,PrRP 受体主要定位于 FW 和 SW 鲶鱼垂体的 RPD 和神经垂体细胞中。我们还检查了 PrRP 对 FW 鲶鱼原代孵育 PRL 细胞中 PRL 表达的影响;PrRP 未能刺激体外 PRL 细胞中 PRL 的表达。这些结果表明,在鱼类中报道的 PrRP 对 PRL 基因表达的体内刺激作用可能是由表达 PrRP 受体的神经垂体分泌的未知调节剂介导的。