Kim Wan-Young, Jung Joon-Ho, Park Eun-Young, Yang Chul-Woo, Kim Hyang, Nielsen Søren, Madsen Kirsten M, Kim Jin
Dept. of Anatomy and MRC for Cell Death Disease Research Center, College of Medicine, The Catholic Univ. of Korea, Seoul, Korea, 505, Banpo-Dong, Seocho-Ku, Seoul 137-701, Korea.
Am J Physiol Renal Physiol. 2006 Nov;291(5):F1052-60. doi: 10.1152/ajprenal.00016.2006. Epub 2006 May 30.
Recent studies of the distribution of PKC isoenzymes in the mouse kidney demonstrated that PKC-alpha, -beta(I), and -delta are expressed in intercalated cells. The purpose of this study was to identify the intercalated cell subtypes that express the different PKC isoenzymes and determine the location of the PKC isoenzymes within these cells. Adult C57BL/6 mice kidney tissues were processed for multiple-labeling immunohistochemistry. Antibodies against the vacuolar H(+)-ATPase and pendrin were used to identify intercalated cell subtypes, whereas antibodies against calbindin D(28K) and aquaporin-2 (AQP2) were used to identify connecting tubule cells and principal cells of the collecting duct, respectively. Within type A intercalated cells, PKC-delta was highly expressed in the apical part of the cells, whereas immunoreactivity for both PKC-alpha and PKC-beta(I) was weak. Type B intercalated cells exhibited strong expression of PKC-alpha, -beta(I), and -delta. PKC-alpha and -beta(I) were localized throughout the cytoplasm, whereas PKC-delta was restricted to the basal domain. Within non-A-non-B cells, immunoreactivity for both PKC-alpha and PKC-beta(I) was high in intensity and localized diffusely in the cytoplasm, whereas PKC-delta was localized in the apical part of the cells. None of the PKC isoenzymes (PKC-alpha, -beta(I), or -delta) were expressed in the calbindin D(28K)-positive connecting tubule cells. Within AQP2-positive principal cells of the collecting duct, PKC-alpha was expressed on the basolateral plasma membrane, but no significant staining was detected for PKC-beta(I) and -delta. In summary, this study demonstrates distinct and differential expression patterns of PKC-alpha, -beta(I), and -delta in the three subtypes of intercalated cells in the mouse kidney.
最近对蛋白激酶C(PKC)同工酶在小鼠肾脏中分布的研究表明,PKC-α、-β(I)和-δ在闰细胞中表达。本研究的目的是确定表达不同PKC同工酶的闰细胞亚型,并确定这些细胞内PKC同工酶的位置。对成年C57BL/6小鼠肾脏组织进行多重标记免疫组织化学处理。针对液泡H(+)-ATP酶和pendrin的抗体用于识别闰细胞亚型,而针对钙结合蛋白D(28K)和水通道蛋白-2(AQP2)的抗体分别用于识别连接小管细胞和集合管主细胞。在A型闰细胞内,PKC-δ在细胞顶端高度表达,而PKC-α和PKC-β(I)的免疫反应性较弱。B型闰细胞表现出PKC-α、-β(I)和-δ的强表达。PKC-α和-β(I)定位于整个细胞质,而PKC-δ局限于基底结构域。在非A非B细胞内,PKC-α和PKC-β(I)的免疫反应性强度高且在细胞质中弥漫性定位,而PKC-δ定位于细胞顶端。在钙结合蛋白D(28K)阳性的连接小管细胞中未表达任何PKC同工酶(PKC-α、-β(I)或-δ)。在集合管的AQP2阳性主细胞内,PKC-α表达于基底外侧质膜,但未检测到PKC-β(I)和-δ的明显染色。总之,本研究证明了PKC-α、-β(I)和-δ在小鼠肾脏闰细胞的三种亚型中存在不同的表达模式。