Minuth W W, Lauer G, Bachmann S, Kriz W
Histochemistry. 1984;80(2):171-82. doi: 10.1007/BF00679993.
A sulfated, proline-rich glycoprotein (gpCDI, apparent molecular weight 200,000 in column chromatography and 150,000 in SDS-PAGE) was isolated from cultured renal collecting duct epithelium by centrifugation. Triton X100 extraction and DEAE-cellulose ion exchange chromatography. A DEAE-cellulose ion exchange chromatography fraction with the enriched gpCDI was used for immunization of guinea pigs. The antiserum was prepared for antigen localization by indirect immunofluorescence in collecting duct cell cultures and in tissue sections of neonatal and adult rabbit kidneys. In the cultured collecting duct epithelium, antibody staining of the epithelium and structures of the extracellular matrix was age dependent. Cultures of dedifferentiated collecting duct monolayers revealed positive reaction in the cytoplasm. In neonatal and adult rabbit kidneys, the antibody was localized in the entire collecting duct system but not in the collecting duct ampullae of the newborn kidney. Staining of the cytoplasm was found only in medullary collecting ducts of the neonatal kidney; other portions revealed staining mostly at the basal circumference of the tubule and at the luminal cell borders. Apart from collecting ducts, no other tubular segments were reactive. The cortical and the medullary interstitium contained fluorescent fibres which were concentrated around vascular structures. A possible relation between gpCDI and collagenous compounds is discussed. Bowman's capsule reacted positively, whereas staining of the mesangial matrix was weak. The localization of the antigen, as revealed by indirect immunofluorescence, suggests that gpCDI occurs both in intracellular and extracellular (interstitial) location. Two main points are emphasized: Firstly gpCDI is considered an important constituent in different stages of collecting duct development, and secondly, the staining pattern of the antibody varies with the different portions of both young and adult kidney collecting ducts; this staining heterogeneity may correspond with the known regional differences of collecting duct functions.
通过离心、Triton X100提取和DEAE - 纤维素离子交换色谱法，从培养的肾集合管上皮细胞中分离出一种硫酸化、富含脯氨酸的糖蛋白（gpCDI，在柱色谱中表观分子量为200,000，在SDS - PAGE中为150,000）。将富含gpCDI的DEAE - 纤维素离子交换色谱级分用于豚鼠免疫。制备抗血清，通过间接免疫荧光法在集合管细胞培养物以及新生和成年兔肾组织切片中进行抗原定位。在培养的集合管上皮细胞中，上皮细胞和细胞外基质结构的抗体染色具有年龄依赖性。去分化的集合管单层细胞培养物在细胞质中显示出阳性反应。在新生和成年兔肾中，抗体定位于整个集合管系统，但不在新生肾的集合管壶腹。仅在新生肾的髓质集合管中发现细胞质染色；其他部分的染色大多位于肾小管的基底圆周和管腔细胞边界处。除集合管外，没有其他肾小管段有反应。皮质和髓质间质含有围绕血管结构集中的荧光纤维。讨论了gpCDI与胶原化合物之间可能的关系。鲍曼囊呈阳性反应，而系膜基质的染色较弱。间接免疫荧光显示的抗原定位表明，gpCDI存在于细胞内和细胞外（间质）位置。强调了两个要点：第一，gpCDI被认为是集合管发育不同阶段的重要组成部分；第二，抗体的染色模式因幼龄和成年肾集合管的不同部分而异；这种染色异质性可能与已知的集合管功能区域差异相对应。
