Huang Z, Ritter C, Brown A, Finch J, Abu-Amer Y, Ross P, Slatopolsky E
Department of Medicine, Renal Division, Washington University School of Medicine, Saint Louis, Missouri, 63110, USA.
Biochem Biophys Res Commun. 1999 Feb 24;255(3):645-51. doi: 10.1006/bbrc.1999.0226.
Rab3 proteins are small GTP-binding proteins known to play a role in regulated exocytosis processes. This study examines the expression of Rab3 mRNA and protein in bovine, rat and human parathyroid glands. mRNAs of several Rab3 isoforms were detected in bovine (Rab3A, Rab3B and Rab3C) and rat (Rab3A, Rab3B and Rab3D) parathyroid glands by RT-PCR and sequencing. Rab3A protein was detected in the cytosolic extract from bovine parathyroid gland by Western blotting using a monoclonal antibody for Rab3A. Rab3A protein was localized to parathyroid hormone-containing chief cells by immunohistochemical staining. Subcellular localization of Rab3A protein by immunogold electron microscopy revealed that the majority of Rab3A protein was not associated with dense-core vesicles, but localized in the cytosol of the chief cells. Altogether, our results demonstrate that Rab3 isoforms are expressed in parathyroid chief cells, suggesting that they may play a role in regulated exocytosis in these cells.
Rab3蛋白是一类已知在调节性胞吐过程中发挥作用的小GTP结合蛋白。本研究检测了Rab3 mRNA和蛋白在牛、大鼠和人甲状旁腺中的表达情况。通过逆转录聚合酶链反应(RT-PCR)和测序,在牛(Rab3A、Rab3B和Rab3C)和大鼠(Rab3A、Rab3B和Rab3D)甲状旁腺中检测到了几种Rab3亚型的mRNA。使用针对Rab3A的单克隆抗体,通过蛋白质免疫印迹法在牛甲状旁腺的胞质提取物中检测到了Rab3A蛋白。通过免疫组织化学染色,Rab3A蛋白定位于含甲状旁腺激素的主细胞。通过免疫金电子显微镜对Rab3A蛋白进行亚细胞定位显示,大多数Rab3A蛋白不与致密核心囊泡相关,而是定位于主细胞的胞质溶胶中。总之,我们的结果表明Rab3亚型在甲状旁腺主细胞中表达,提示它们可能在这些细胞的调节性胞吐中发挥作用。