大鼠肾小球冷冻断裂细胞化学研究

应用断裂冷冻法体外标记肾小球成份,定位WGA受体和阴电荷点。组织经冷冻断裂和冻融后,标记物无阻碍地到达细胞表面和基质横断面可直接比较结合活性,使用的示踪物可以不管缓冲液的pH,用阳离子铁蛋白(CF,PI 8.3)测定肾小球阴电荷点,所有细胞表面、基底膜和系膜基质断裂面呈连续标记,在所有细胞表面和断裂基质见到WGA—金胶复合物。神经氨酸酶处理后,WGA不再与足细胞和基底膜结合,但与内皮细胞、系膜细胞和系膜基质断裂面结合,提示在足细胞WGA结合NeuNAc,在内皮细胞、系膜细胞和系膜基质结合GIcNAc,在基底膜结合NeuNAc和GIcNAc。

Freeze fracture Cytochemistry Study of Rat Glomerulus

The freeze-fracture was used to gain access and to label in vitro g(?)omeru(?)ar components and locate WGA receptors and anionic sites. Tissues are frozen. fractured under liquid nitrogen and thawed. Unrestricted access of label to cell surfaces and cross-fractured matrices allows a direct comparison of binding activities and probes can be used regardless the pH of the buffers. The glomerular anionic charges are determined with cationized ferritin (CF. PI 8.3), Continuous CF labeling was seen all over the cell surfaces and cross-fracture profiles of GBM and mesangial matrix (MM). WGA-gold complexes were observed on all cell surfaces and fractured matrices. After handling with neuraminidase, WGA no longer bound to podocytes and GBM, but it labeled endothelial and mesangial cells and cross-fracture profiles of MM. WGA might probably bind to NeuNAc in podocytes, GLcNAc in endothelial and mesangial cells and MM, and to both NeuNAc and GlcNAc in GBM.