Kloor D, Stumvoll W, Schmid H, Kömpf J, Mack A, Osswald H
Department of Pharmacology, Faculty of Medicine, University of Tübingen, Germany.
J Histochem Cytochem. 2000 Feb;48(2):211-8. doi: 10.1177/002215540004800206.
S-adenosylhomocysteine (SAH) hydrolase is a cytosolic enzyme present in the kidney. Enzyme activities of SAH hydrolase were measured in the kidney in isolated glomeruli and tubules. SAH hydrolase activity was 0.62 +/- 0.02 mU/mg in the kidney, 0.32 +/- 0.03 mU/mg in the glomeruli, and 0.50 +/- 0.02 mU/mg in isolated tubules. Using immunohistochemical methods, we describe the localization of the enzyme SAH hydrolase in rat kidney with a highly specific antibody raised in rabbits against purified SAH hydrolase from bovine kidney. This antibody crossreacts to almost the same extent with the SAH hydrolase from different species such as rat, pig, and human. Using light microscopy, SAH hydrolase was visualized by the biotin-streptavidin-alkaline phosphatase immunohistochemical procedure. SAH hydrolase immunostaining was observed in glomeruli and in the epithelium of the proximal and distal tubules. The collecting ducts of the cortex and medulla were homogeneously stained. By using double immunofluorescence staining and two-channel immunofluorescence confocal laser scanning microscopy, we differentiated the glomerular cells (endothelium, mesangium, podocytes) and found intensive staining of podocytes. Our results show that the enzyme SAH hydrolase is found ubiquitously in the rat kidney. The prominent staining of SAH hydrolase in the podocytes may reflect high rates of transmethylation. (J Histochem Cytochem 48:211-218, 2000)
S-腺苷同型半胱氨酸(SAH)水解酶是一种存在于肾脏中的胞质酶。在分离出的肾小球和肾小管中测定了肾脏中SAH水解酶的酶活性。肾脏中SAH水解酶活性为0.62±0.02 mU/mg,肾小球中为0.32±0.03 mU/mg,分离出的肾小管中为0.50±0.02 mU/mg。我们使用免疫组织化学方法,用兔抗牛肾纯化SAH水解酶产生的高特异性抗体描述了SAH水解酶在大鼠肾脏中的定位。该抗体与大鼠、猪和人等不同物种的SAH水解酶几乎以相同程度发生交叉反应。使用光学显微镜,通过生物素-链霉亲和素-碱性磷酸酶免疫组织化学程序观察到SAH水解酶。在肾小球以及近端和远端肾小管的上皮中观察到SAH水解酶免疫染色。皮质和髓质的集合管被均匀染色。通过使用双重免疫荧光染色和双通道免疫荧光共聚焦激光扫描显微镜,我们区分了肾小球细胞(内皮细胞、系膜细胞、足细胞),并发现足细胞有强烈染色。我们的结果表明,SAH水解酶在大鼠肾脏中普遍存在。足细胞中SAH水解酶的显著染色可能反映了高甲基化率。(《组织化学与细胞化学杂志》48:211 - 218,2000)