Hnasko R, McFarland M, Ben-Jonathan N
Department of Cell Biology, Neurobiology and Anatomy, University of Cincinnati Medical School, 3125 Eden Ave, Cincinnati, Ohio 45267, USA.
J Endocrinol. 2002 Dec;175(3):649-61. doi: 10.1677/joe.0.1750649.
Plasmalemma vesicle protein-1 (PV-1) is an integral membrane protein associated with endothelial cell caveolae and fenestrae. Since endocrine glands are enriched with fenestrated endothelium, we examined the distribution of PV-1 mRNA and protein in endocrine glands and determined its cellular localization. A single transcript was detected by RT-PCR in all endocrine glands examined. A synthetic peptide was used to generate antibodies for Western blotting and immunohistochemistry (IHC). Western blotting of membrane fractions from lung, pituitary, adrenal, testis and PV-1-transfected Cos-1 cells revealed a major 65 kDa protein. This protein binds to heparin with high affinity. Using IHC, PV-1 was localized to both endothelial cells of the adrenal zona reticularis and chromaffin cells of the medulla. In the pancreas, PV-1 expression was restricted to a few cells in the islets of Langerhans that partially overlap with somatostatin-positive delta-cells. In both neonatal and adult pituitaries, strong PV-1 immunoreactivity was detected in neural lobe pituicytes in a pattern similar to that of glial fibrillary acidic protein (GFAP). PV-1 and GFAP expression was seen in the adult, but not neonatal, intermediate lobe. Endothelial cells throughout the neonatal anterior lobe were PV-1 positive, but PV-1 in the adult was restricted to some endothelial and endocrine cells localized near the margins of lobe. In the adult testis, strong PV-1 expression was seen in germ cells within the seminiferous tubules that varied with the stage of spermatogenesis. In contrast, PV-1 in the neonatal testis was localized to the interstitial cells but not seminiferous tubules. In the ovary, PV-1 was expressed in stromal endothelial cells as well as the thecal layer of developing follicles. Over half the corpus luteal cells were positive for PV-1. Our data have shown that PV-1 is not restricted to endothelial cells but is localized in many types of endocrine and non-endocrine cells. Furthermore, PV-1 expression in the pituitary and testis is developmentally regulated.
质膜囊泡蛋白-1(PV-1)是一种与内皮细胞小窝和窗孔相关的整合膜蛋白。由于内分泌腺富含窗孔状内皮,我们检测了PV-1 mRNA和蛋白在内分泌腺中的分布,并确定了其细胞定位。通过RT-PCR在所有检测的内分泌腺中检测到单一转录本。使用合成肽制备抗体用于蛋白质印迹和免疫组织化学(IHC)。对来自肺、垂体、肾上腺、睾丸和PV-1转染的Cos-1细胞的膜组分进行蛋白质印迹分析,发现一种主要的65 kDa蛋白。该蛋白与肝素具有高亲和力。使用免疫组织化学方法,PV-1定位于肾上腺网状带的内皮细胞和髓质的嗜铬细胞。在胰腺中,PV-1表达局限于胰岛中的少数细胞,这些细胞与生长抑素阳性的δ细胞部分重叠。在新生儿和成人垂体中,在神经叶垂体细胞中检测到强烈的PV-1免疫反应性,其模式类似于胶质纤维酸性蛋白(GFAP)。PV-1和GFAP表达见于成人垂体中间叶,但不见于新生儿垂体中间叶。新生儿前叶的内皮细胞均为PV-1阳性,但成人垂体中的PV-1局限于叶边缘附近的一些内皮细胞和内分泌细胞。在成人睾丸中,在生精小管内的生殖细胞中可见强烈的PV-1表达,其表达随精子发生阶段而变化。相比之下,新生儿睾丸中的PV-1定位于间质细胞而非生精小管。在卵巢中,PV-1在基质内皮细胞以及发育卵泡的卵泡膜层中表达。超过一半的黄体细胞PV-1呈阳性。我们的数据表明,PV-1不仅局限于内皮细胞,还定位于多种内分泌和非内分泌细胞。此外,垂体和睾丸中PV-1的表达受发育调控。