Developmental schedule of the postnatal rat testis determined by flow cytometry

Abstract

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.