Morales C, Sylvester S R, Griswold M D
Program in Biochemistry, Washington State University, Pullman 99164.
Biol Reprod. 1987 Nov;37(4):995-1005. doi: 10.1095/biolreprod37.4.995.
The transport of radioactive iron across the seminiferous tubules was analyzed in vivo by light-microscope quantitative radioautography. At 5 min after a single intratesticular injection of 55Fe-transferrin, a strong labeling of the basal aspect of the seminiferous epithelium was observed. Between 30 min and 2 h, the labeling on the basal aspect of the seminiferous epithelium decreased. This decrease was accompanied by a substantial increase of the radioautographic reaction over the cellular elements in the adluminal compartment. These results were consistent with the demonstration of 59Fe associated with meiotic spermatocytes and differentiating spermatids isolated by velocity sedimentation from testes injected with 59Fe-transferrin. Furthermore, after a single intratesticular injection of 59Fe-labeled human transferrin, radiolabeled rat transferrin was immunoprecipitated from homogenates of isolated tubules with a specific antibody and appeared as a single radioactive band on fluorographs of urea/polyacrylamide gels. Similarly, 59Fe-labeled rat transferrin but not 125I-transferrin was immunoprecipitated from rete testis fluids of testes infused with either 59Fe- or 125I-labeled human transferrin. Finally, the synthesis of testicular transferrin in vivo was demonstrated in fluorographs of immunoprecipitated transferrin after an intratesticular injection of 35S-methionine in rats whose livers were excluded from the general circulation by ligation of both the hepatic artery and the portal vein. Thus, our results demonstrated a unidirectional system of iron transport from the basal compartment of the seminiferous epithelium to the germ cells in the adluminal compartment involving two distinct transferrins, i.e., a serum transferrin and a testicular transferrin synthesized by the seminiferous epithelium.
通过光学显微镜定量放射自显影技术对放射性铁在体内穿过生精小管的运输情况进行了分析。在睾丸内单次注射55Fe-转铁蛋白后5分钟,观察到生精上皮基底部有强烈的标记。在30分钟至2小时之间,生精上皮基底部的标记减少。这种减少伴随着近腔室细胞成分上放射自显影反应的显著增加。这些结果与通过速度沉降从注射了59Fe-转铁蛋白的睾丸中分离出的减数分裂精子细胞和分化中的精子细胞中59Fe的证明一致。此外,在睾丸内单次注射59Fe标记的人转铁蛋白后,用特异性抗体从分离的小管匀浆中免疫沉淀出放射性标记的大鼠转铁蛋白,并且在尿素/聚丙烯酰胺凝胶的荧光照片上呈现为单一放射性条带。同样,从注入了59Fe-或125I标记的人转铁蛋白的睾丸的睾丸网液中免疫沉淀出59Fe标记的大鼠转铁蛋白,而不是125I-转铁蛋白。最后,在通过结扎肝动脉和门静脉将肝脏排除在体循环之外的大鼠中,在睾丸内注射35S-甲硫氨酸后,在免疫沉淀的转铁蛋白的荧光照片中证明了体内睾丸转铁蛋白的合成。因此,我们的结果证明了一个铁运输的单向系统,即从生精上皮的基底部到近腔室中的生殖细胞,涉及两种不同的转铁蛋白,即血清转铁蛋白和由生精上皮合成的睾丸转铁蛋白。