Sylvester S R, Griswold M D
Biol Reprod. 1984 Aug;31(1):195-203. doi: 10.1095/biolreprod31.1.195.
One of the major proteins secreted by rat Sertoli cells in culture is a transferrin-like protein (Skinner and Griswold, 1980). The purpose of this study was to quantitate the amount of testicular transferrin in fluids isolated from the testis by the use of a radioimmunoassay and to determine the location of transferrin and transferrin receptors in the testis by indirect immunofluorescence. Seminiferous tubule fluid, rete testis fluid, and testicular lymph were collected from rat testes and were found to contain 141 micrograms, 47 micrograms and 3.7 mg transferrin per ml of fluid, respectively. Serum was found to contain 3.7 mg/ml transferrin. Paraffin sections of rat testis were incubated with rabbit anti-rat transferrin, biotinylated goat anti-rabbit and fluorescein-conjugated avidin. Immunoreactive transferrin was thus localized on the proacrosome and nuclear cap of developing spermatids. Late spermatids showed transferrin over the entire region of the head but mature testicular spermatozoa exhibited little fluorescence. The interstitial tissue between seminiferous tubules fluoresced brightly, indicating a large amount of transferrin in this area. By pretreating sections with rat transferrin, the receptor for the protein was localized on and in spermatocytes and early round spermatids. Dividing germ cells were brightly fluorescent.
培养的大鼠支持细胞分泌的主要蛋白质之一是一种类转铁蛋白(斯金纳和格里斯沃尔德,1980年)。本研究的目的是使用放射免疫分析法对从睾丸分离的液体中的睾丸转铁蛋白量进行定量,并通过间接免疫荧光法确定转铁蛋白和转铁蛋白受体在睾丸中的位置。从大鼠睾丸收集生精小管液、睾丸网液和睾丸淋巴液,发现每毫升液体中分别含有141微克、47微克和3.7毫克转铁蛋白。发现血清中含有3.7毫克/毫升转铁蛋白。将大鼠睾丸石蜡切片与兔抗大鼠转铁蛋白、生物素化山羊抗兔抗体和荧光素偶联抗生物素蛋白一起孵育。免疫反应性转铁蛋白因此定位于发育中的精子细胞的前顶体和核帽上。晚期精子细胞在头部的整个区域都显示有转铁蛋白,但成熟的睾丸精子荧光很少。生精小管之间的间质组织荧光强烈,表明该区域有大量转铁蛋白。通过用大鼠转铁蛋白预处理切片,该蛋白质的受体定位于精母细胞和早期圆形精子细胞上及细胞内。正在分裂的生殖细胞荧光强烈。