Enders G C, Kahsai T Z, Lian G, Funabiki K, Killen P D, Hudson B G
Department of Anatomy and Cell Biology, Kansas University Medical Center, Kansas City 66103, USA.
Biol Reprod. 1995 Dec;53(6):1489-99. doi: 10.1095/biolreprod53.6.1489.
The temporal expression of type IV collagen, laminin, and entactin in the basal laminae of the seminiferous tubule during development of the mouse testis was determined. Northern blot analysis was used to examine changes in testicular mRNA for alpha 1-alpha 5 type IV collagen (IV) chains in mice ranging in age from newborn to adult (60 days). Levels for mRNA alpha 1(IV) and alpha 2(IV) chains were highest in newborns through Day 5 and remained elevated through Day 10, but then sharply declined to adult values by Day 30. In sharp contrast, alpha 3(IV) and alpha 4(IV) chain levels were low in newborns, peaked at Day 10, and then declined to adult values by Day 30. 5(IV) mRNA was elevated in newborns and at Days 5 and 10 before dropping to adult levels by Day 30. Changes in the deposition of alpha 1, alpha 2, and alpha 3(IV) collagen chains, laminin, and entactin into the inner and outer basal laminae of the seminiferous tubule were determined from the beginning of tubule formation (embryonic Day 12.5) through adulthood by immunofluorescence microscopy using polyclonal antibodies for these constituents. The alpha 1 and alpha 2(IV) chains, laminin, and entactin were deposited into the inner basal lamina at embyronic Day 12.5 and into the newly formed outer basal lamina at Day 5 after birth. The alpha 3(IV) chains were deposited into both the inner and outer basal laminae at Day 5. Thus, testicular alpha 1-alpha 3(IV) mRNA levels coincide with the incorporation of detectable collagen chains into the seminiferous basal laminae, suggesting transcriptional control of these alpha (IV) chains. Expression of of the alpha 3(IV) chain coincides with the initiation of spermatogenesis, suggesting a functional role of this chain in spermatogonial proliferation.
测定了小鼠睾丸发育过程中,生精小管基底膜中IV型胶原、层粘连蛋白和巢蛋白的时序表达。采用Northern印迹分析,检测了从新生小鼠到成年小鼠(60日龄)睾丸中α1-α5 IV型胶原(IV)链mRNA的变化。mRNA α1(IV)和α2(IV)链水平在新生小鼠至第5天最高,至第10天仍保持升高,但到第30天急剧下降至成年水平。与之形成鲜明对比的是,α3(IV)和α4(IV)链水平在新生小鼠中较低,在第10天达到峰值,然后到第30天下降至成年水平。5(IV) mRNA在新生小鼠以及第5天和第10天升高,然后到第30天降至成年水平。利用针对这些成分的多克隆抗体,通过免疫荧光显微镜,从生精小管形成开始(胚胎第12.5天)到成年期,测定了α1、α2和α3(IV)胶原链、层粘连蛋白和巢蛋白在生精小管内、外基底膜中的沉积变化。α1和α2(IV)链、层粘连蛋白和巢蛋白在胚胎第12.5天沉积到内基底膜中,并在出生后第5天沉积到新形成的外基底膜中。α3(IV)链在第5天沉积到内、外基底膜中。因此,睾丸α1-α3(IV) mRNA水平与可检测到的胶原链掺入生精小管基底膜的过程一致,表明这些α(IV)链存在转录调控。α3(IV)链的表达与精子发生的起始一致,表明该链在精原细胞增殖中具有功能作用。