Huang H F
Department of Surgery, UMD-New Jersey Medical School, Newark 07103.
Biol Reprod. 1994 Jul;51(1):24-33. doi: 10.1095/biolreprod51.1.24.
The aim of the present study was to examine the feasibility of inducing enrichment of the stages of the seminiferous epithelium in prepubertal male rats produced by vitamin A-deficient (VAD) females that had received a supplement of a marginal level of retinol. Seventy-day-old retinoic acid-supplemented VAD female rats were given daily doses of 5, 10, or 20 micrograms retinol for 2 wk. Breeding of these females resulted in the production of viable offspring in litters of normal size. Subsequently, male rats so produced (VAD males) were given a single injection of 10 or 20 micrograms retinol at 10 or 20 days of age, fostered by normal lactating females, and weaned at 25 days of age. These rats were maintained on normal rat chow thereafter. At 10 days of age, active spermatogonial proliferation was not seen in the testes of VAD males born to VAD females receiving a 5- or 10-micrograms daily dose of retinol. Concurrently, cells resembling gonocytes were frequently seen, and pre-type A spermatogonia were seen in over 80% of the tubules examined. By 20 days of age, the presence of active spermatogonial proliferation was demonstrated by the appearance of preleptotene spermatocytes, but these failed to differentiate further. Spermatogenesis of the VAD males born to VAD females given a daily dose of 20 micrograms retinol appeared to be normal at both 10 and 20 days of age. Forty days after the retinol replacement, the testes of VAD males born to VAD females given a daily dose of 5 or 10 micrograms retinol were enriched with specific stages of the seminiferous epithelium while other stages were absent. The stages enriched were affected by the dose of retinol administered and the age when retinol replacement began. The stage enrichment in these animals persisted for at least 135 days. Seminiferous epithelium of the VAD males born to the VAD females receiving a 20-micrograms daily dose of retinol was not stage-enriched. Northern blot analysis of testicular RNA revealed stage-dependent changes in the steady-state level of mRNA for spermatid protamine and Sertoli cell androgen-binding protein (ABP). Results of this study demonstrate the feasibility of inducing enrichment of seminiferous stages in prepubertal rats. The production of this animal model is cost-effective in that it saves in time and animal maintenance.(ABSTRACT TRUNCATED AT 400 WORDS)
本研究的目的是检验通过给维生素A缺乏(VAD)的雌性大鼠补充微量视黄醇来诱导青春期前雄性大鼠生精上皮阶段富集的可行性。给70日龄补充视黄酸的VAD雌性大鼠每日分别给予5、10或20微克视黄醇,持续2周。这些雌性大鼠繁殖后产出了正常大小窝的存活后代。随后,将如此产生的雄性大鼠(VAD雄性大鼠)在10或20日龄时单次注射10或20微克视黄醇,由正常哺乳的雌性大鼠哺育,并在25日龄时断奶。此后这些大鼠一直喂以正常大鼠饲料。在10日龄时,接受每日5或10微克视黄醇剂量的VAD雌性大鼠所生VAD雄性大鼠的睾丸中未见活跃的精原细胞增殖。同时,经常可见类似生殖母细胞的细胞,在所检查的超过80%的曲细精管中可见A型前精原细胞。到20日龄时,前细线期精母细胞的出现证明有活跃的精原细胞增殖,但这些细胞未能进一步分化。每日给予20微克视黄醇的VAD雌性大鼠所生VAD雄性大鼠在10日龄和20日龄时精子发生似乎均正常。视黄醇替代40天后,接受每日5或10微克视黄醇剂量的VAD雌性大鼠所生VAD雄性大鼠的睾丸富含生精上皮的特定阶段,而其他阶段则缺失。富集的阶段受所给予视黄醇剂量和视黄醇替代开始时的年龄影响。这些动物中的阶段富集持续了至少135天。接受每日20微克视黄醇剂量的VAD雌性大鼠所生VAD雄性大鼠的生精上皮未出现阶段富集。睾丸RNA的Northern印迹分析显示,精子细胞鱼精蛋白和支持细胞雄激素结合蛋白(ABP)的mRNA稳态水平存在阶段依赖性变化。本研究结果证明了在青春期前大鼠中诱导生精阶段富集的可行性。该动物模型的产生具有成本效益,因为它节省了时间和动物饲养成本。(摘要截短至400字)