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Expression of microfibrillar proteins by bovine bladder urothelium.

机译:牛膀胱尿路上皮表达微纤维蛋白。

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OBJECTIVES: To determine the occurrence and potential function of proteins composing elastic microfibrils in the developing bovine bladder. METHODS: Monospecific antibodies, generated against two well-characterized microfibrillar proteins, microfibril-associated glycoprotein (MAGP) and fibrillin-1 (FBN1), were used in immunohistochemical analysis of full-thickness frozen sections of fetal bovine bladder. The localization of these two antibodies was compared with that of anti-type IV collagen antibody. Adjacent serial sections were stained for routine light microscopy. Cultured urothelial cells were fixed in 3.7% formaldehyde and permeabilized with 0.5% Triton X-100 before immunoanalysis. Control reactions used either preimmune serum or a monoclonal antibody to a nonmatrix protein. Poly(A+) ribonucleic acid was isolated from cultured urothelial cells and subjected to Northern analysis using specific complementary deoxyribonucleic acid probes for MAGP and FBN1. RESULTS: Both MAGP and FBN1 are expressed by the urothelium and are found in association with the underlying basement membrane, as visualized by their co-localization with type IV collagen. Furthermore, urothelial cells in culture continue to express both microfibrillar proteins. CONCLUSIONS: The developing bovine urothelium expresses major microfibrillar protein components. The role of these microfibrils in the urothelium remains to be determined, but they may have an important anchoring function.
机译:目的:确定在发育中的牛膀胱中构成弹性微纤维的蛋白质的发生和潜在功能。方法:针对两种特征明确的微纤维蛋白,微纤维相关糖蛋白(MAGP)和原纤维蛋白1(FBN1)产生的单特异性抗体用于免疫组化分析胎儿牛膀胱全层冰冻切片。将这两种抗体的定位与抗IV型胶原抗体的定位进行比较。将相邻的连续切片染色以进行常规光学显微镜检查。在免疫分析之前,将培养的尿路上皮细胞固定在3.7%的甲醛中,并用0.5%的Triton X-100透化。对照反应使用免疫前血清或针对非基质蛋白的单克隆抗体。从培养的尿路上皮细胞中分离出聚(A +)核糖核酸,并使用针对MAGP和FBN1的特异性互补脱氧核糖核酸探针进行Northern分析。结果:MAGP和FBN1均由尿路上皮表达,并与下层基底膜相关联,通过与IV型胶原蛋白共定位可见。此外,培养物中的尿道上皮细胞继续表达两种微纤维蛋白。结论:正在发育的牛尿路上皮表达主要的微纤维蛋白成分。这些微纤维在尿路上皮中的作用尚待确定,但它们可能具有重要的锚固功能。

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