Malkov M, Fisher Y, Don J
Department of Life Sciences, Bar-Ilan University, Ramat-Gan, Israel.
Biol Reprod. 1998 Jul;59(1):84-92. doi: 10.1095/biolreprod59.1.84.
Analysis of the biochemical events and the genes expressed at various postnatal developmental stages in the testis of mammals is of great importance for understanding spermatogenesis in general and meiosis in particular. A prerequisite for such an analysis is the characterization of a detailed developmental schedule of the postnatal testis. In this study we used four-parameter flow cytometry analysis to determine a detailed testicular developmental schedule in rats as compared to mice. A dot plot of forward-scatter/side-scatter of testicular cell suspensions from mature animals revealed 7 distinct subpopulations within the testis. These, when analyzed by fluorescence parameters, were divided into 4 levels of fluorescence: cells containing 4d DNA, 2d DNA, and 2 levels of haploid cells. Observing the acquisition pattern of these subpopulations during postnatal development, we were able to suggest the following developmental schedule for the rat. At postnatal Days 6-7, the testis contains somatic cells and spermatogonia cells only. By Days 13-14, leptotene spermatocytes appear; by Days 17-18, zygotene spermatocytes are present; by Days 19-20 and Days 22-23, early and late pachytene spermatocytes, respectively, are seen. Haploid round spermatids first appear at Days 24-25 and elongating spermatids by Days 30-31; by Day 36, elongated spermatozoa can be found.
分析哺乳动物睾丸在出生后各个发育阶段发生的生化事件和表达的基因,对于全面理解精子发生,尤其是减数分裂,具有重要意义。进行此类分析的一个前提是明确出生后睾丸详细的发育进程。在本研究中,我们使用四参数流式细胞术分析来确定大鼠与小鼠相比详细的睾丸发育进程。来自成熟动物的睾丸细胞悬液的前向散射/侧向散射点图显示睾丸内有7个不同的亚群。通过荧光参数分析,这些亚群被分为4种荧光水平:含有4倍体DNA的细胞、2倍体DNA的细胞以及2种单倍体细胞水平。观察这些亚群在出生后发育过程中的获得模式,我们能够提出以下大鼠的发育进程。在出生后第6 - 7天,睾丸仅包含体细胞和精原细胞。到第13 - 14天,细线期精母细胞出现;到第17 - 18天,偶线期精母细胞出现;到第19 - 20天和第22 - 23天,分别可见早期和晚期粗线期精母细胞。单倍体圆形精子细胞最早在第24 - 25天出现,伸长的精子细胞在第30 - 31天出现;到第36天,可以发现伸长的精子。