Kutsuna Miki, Kodama Toshio, Sumida Michihiro, Nagai Ayako, Higashine Masakatsu, Zhang Wei, Hayashi Yasuhito, Shiraishi Atsushi, Ohashi Yuichi
Department of Ophthalmology, School of Medicine, Ehime University, Shitsukawa, Toon-City, Ehime 791-0295, Japan.
Exp Eye Res. 2007 Apr;84(4):687-93. doi: 10.1016/j.exer.2006.12.007. Epub 2006 Dec 15.
Adipose differentiation-related protein (ADRP) is an intrinsic lipid storage protein found in lipid droplets of different type of cells. ADRP has been recognized to be a specific marker of lipid accumulation and a marker of differentiated adipocytes. The purpose of this study was to determine whether ADRP was present in the cells of the meibomian gland. The expression of the mRNA of ADRP was determined by RT-PCR and Northern blot analysis of the meibomian gland and other rat tissues. A newly generated polyclonal antibody against rat ADRP was used for Western blot analysis and immunohistochemical staining to determine whether ADRP was expressed in the rat meibomian gland. Meibomian gland acinar cells were isolated to determine when ADRP was expressed during cell differentiation in vitro. Northern blot analysis and Western analysis showed that ADRP was expressed in the meibomian gland. Immunoreactivity to ADRP was observed in the lobules of acinar cells in the meibomian gland, and was preferentially located adjacent the vacuolated cytoplasm. In culture, the meibocytes began to store lipid droplets in the cytoplasm as they became confluent, and the immunoreactivity for ADRP was found at the margins of the oil droplets. Our results suggest that ADRP can serve as a new marker for the identification of differentiated meibocytes containing lipid droplets.
脂肪分化相关蛋白(ADRP)是一种存在于不同类型细胞脂滴中的内在脂质储存蛋白。ADRP已被公认为脂质积累的特异性标志物和分化脂肪细胞的标志物。本研究的目的是确定睑板腺细胞中是否存在ADRP。通过对睑板腺和其他大鼠组织进行逆转录聚合酶链反应(RT-PCR)和Northern印迹分析来测定ADRP的mRNA表达。使用新制备的针对大鼠ADRP的多克隆抗体进行蛋白质印迹分析和免疫组织化学染色,以确定ADRP是否在大鼠睑板腺中表达。分离睑板腺腺泡细胞以确定ADRP在体外细胞分化过程中的表达时间。Northern印迹分析和蛋白质印迹分析表明ADRP在睑板腺中表达。在睑板腺腺泡细胞小叶中观察到对ADRP的免疫反应性,并且优先位于空泡化细胞质附近。在培养过程中,睑板细胞汇合时开始在细胞质中储存脂滴,并且在油滴边缘发现了对ADRP的免疫反应性。我们的结果表明,ADRP可作为一种新的标志物,用于鉴定含有脂滴的分化睑板细胞。