ABSTRACT
Extracellular matrix (ECM)is not only one of current hot-points in medical cell biology,but also one tricky part for undergraduates to learn. This article compared chapters of ECM in medical cell biology courses and spotlighted that ECM chapter was often neglected in some domes-tic universities. Then it analyzed the possible causes,such as variable arrangement of ECM section in current textbooks. Lastly,the article recommended several suggestions,including giving a timely re-vision of the old ECM knowledge,designing an appropriate strategy for teaching,enumerating certain representative diseases to improve the ECM education. It appealed our teachers to pay more attention to how to make the module of ECM master well in near future.
ABSTRACT
Objective To prepare and identify monoclonal antibody against LRR-WSC domain of polycystin-1 and to investigate the distribution of polycystin-1 in kidney tissues and kidney cell lines. Methods BALB/c mice were immunized with fusion protein PC1-e of polycystin-1 LRR-WSC domain. The splenocytes were fused with myeloma cells by PEG 4000 and the hybridomas were selected in HAT medium. The hybridoma clones secreting antibodies against polycystin-1 LRR-WSC domain were detected by enzyme-linked immunosorbent assay ( ELISA) and cloned by limiting dilution. The specificity of anti-polycystin-1 LRR-WSC domain monoclonal antibody from hybridoma was verified by ELISA and Western blot. The distribution of polycystin-1 in tissues and cells was detected by immunohistochemical method. Results One cell line of hybridoma secreting monoclonal antibody against polycystin-1 was established. Western blot analysis showed that the monoclonal antibody reacted strongly and specifically to polycystin-1 LRR-WSC domain. Distribution of polycystin-1 in fetal kidney was localized in tubular epithelium. In normal adult kidney tissues, our study showed that polycystin-1 was mainly expressed in the medullary collecting ducts and distal convoluted tubules. Positive staining was also found in the majority of cyst-lining epithelial ceEs of cystic tissue from autosomal dominant polycystic kidney disease ( ADPKD) patients. Expressions of polycystin-1 were found in either ADPKD cyst-lining epithelia cell line and LLC-PK1, clearly plasma membrane and intracytoplasmic staining of polycystin-1 were observed. Conclusion Specific monoclonal antibody against polycystin-1 LRR-WSC domain were obtained. The antibody is important to researching the mechanism of ADPKD. The distribution of polycystin-1 in kidney tissues and cells show that polycystin-1 was important in tubular elongation and the maintenance of tubular architecture.