Tojo Akihiro, Hatakeyama Saaya, Kinugasa Satoshi, Fukuda Satoru, Sakai Toshio
Division of Nephrology and Endocrinology, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8655, Japan.
Laboratory of Electron Microscopy, The University of Tokyo, Tokyo, Japan.
Med Mol Morphol. 2017 Jun;50(2):86-93. doi: 10.1007/s00795-016-0151-6. Epub 2017 Mar 17.
Albumin endocytosis is enhanced in the podocytes of minimal change nephrotic syndrome. We investigated that the endocytic vesicle transport in the podocyte using three-dimensional observation in a rat model of puromycin aminonucleoside (PAN)-induced nephrotic syndrome. At day 7, Evans Blue-labeled albumin was intravenously injected in PAN rats, and one kidney was fixed for a morphological analysis; the other was used for the isolation of glomeruli through sieving and protein analyses. Evans Blue-labeled albumin was found to accumulate in an increased number of vesicles in the podocytes of PAN rat. Continuous sections and its three-dimensional observation demonstrated that vesicles may be transported from the cytoplasm to the apical membrane of the podocytes. The increased protein bands in the gel electrophoresis of the sieved glomeruli of nephrotic rats were analyzed by mass spectrometry in comparison to the control rats. The major proteins increased in the nephrotic rats were cytoplasmic dynein 1 heavy chain, myosin IX, and myosin VIIb. In conclusion, the podocyte endocytic vesicles carrying albumin increased with glomerular cytoplasmic dynein and myosin in minimal change nephrotic rats.
微小病变肾病综合征足细胞中的白蛋白内吞作用增强。我们在嘌呤霉素氨基核苷(PAN)诱导的肾病综合征大鼠模型中,利用三维观察法研究了足细胞中的内吞囊泡运输。在第7天,向PAN大鼠静脉注射伊文思蓝标记的白蛋白,一侧肾脏固定用于形态学分析;另一侧肾脏用于通过筛分分离肾小球并进行蛋白质分析。发现伊文思蓝标记的白蛋白在PAN大鼠足细胞中积累在数量增加的囊泡中。连续切片及其三维观察表明,囊泡可能从足细胞的细胞质转运至顶膜。与对照大鼠相比,对肾病大鼠筛分后的肾小球进行凝胶电泳,通过质谱分析增加的蛋白条带。肾病大鼠中增加的主要蛋白质是胞质动力蛋白1重链、肌球蛋白IX和肌球蛋白VIIb。总之,在微小病变肾病大鼠中,携带白蛋白的足细胞内吞囊泡随着肾小球胞质动力蛋白和肌球蛋白而增加。