Interrupted proliferation and stimulated apoptosis promote testis involution after the rut, and testosterone seems to play a role in the regulation of both processes.
The roe deer (Capreolus capreolus) is a seasonal breeder. The cyclic changes between totally arrested and highly activated spermatogenesis offer an ideal model to study basic mechanisms of spermatogenesis. In this study, we demonstrated, to our knowledge for the first time, c-kit receptor-positive cells in the testis of roe deer. They were immunohistologically identified mainly as spermatogonia. Analysis of the amount of those cells by flow cytometry shows a distinct seasonal pattern, with pronounced differences between cells in the diploid state and in the G2/M phase of mitosis. The specific seasonal pattern of spermatogonial proliferation results in the increased relative abundance of spermatogonia as well as in their increased total number per testis in November and December. This suggests that cell divisions continue on a level sufficient to accumulate spermatogonia during winter. The serum concentrations of LH and FSH showed a peak in spring; testosterone showed a maximum concentration during the rut (July/August). The peak of both gonadotropins seems to precede the period of stimulated spermatogonial proliferation in spring. The testosterone peak coincides with maximal meiotic intensity in August. The results suggest the importance of testosterone for sperm production, and they provide a basis for detailed investigations of regulatory factors of the proliferation of spermatogonia.
Teratozoospermia (ejaculation of <40% morphologically normal sperm) commonly occurs within the Felidae, including certain domestic cats, but the cellular and molecular mechanisms that give rise to this phenomenon remain unknown. This study quantified spermatogenesis to identify differential dysfunctions in teratospermic versus normospermic (>60% normal sperm/ejaculate) domestic cats. Sperm used were from electroejaculates and cauda epididymides. Testes from 10 normo- and 10 teratospermic males were obtained by castration and then evaluated by histomorphometry, flow cytometry, and testicular testosterone enzyme immunoassay. Some morphometric traits (tubular diameter, epithelium height, interstitial area, number of Leydig cells, and blood vessels per cross-section) as well as testicular testosterone concentrations were similar between groups, but testicular volume was greater in teratospermic males. Stage frequencies differed also between both cat populations, suggesting possible dysfunctions in spermiation. Quantification of cell populations in most frequent stages revealed more spermatogenic cells and fewer Sertoli cells per tubule cross-section as well as per tissue unit in teratospermic donors. Hence, the ratio of spermatogenic cells per Sertoli cell was elevated in the teratospermic cat. DNA flow cytometry confirmed higher total spermatogenic and meiotic transformations in teratospermic males. In summary, compared with normospermic counterparts, teratospermic cats have a higher sperm output achieved by more sperm-producing tissue, more germ cells per Sertoli cell, and reduced germ cell loss during spermatogenesis. Gains in sperm quantity are produced at the expense of sperm quality.
Roe deer are seasonal breeders and show cyclic variation in testicular volume and cellular differentiation within the tubular and interstitial testis compartment. We have employed the roe deer as a model to elucidate the expression of the postpubertal Leydig cell marker INSL3 during seasonal changes in Leydig cell differentiation. Roe deer testis and serum samples were collected bimonthly throughout the complete reproductive cycle. Peak levels of testicular Insl3 mRNA and INSL3 immunoprotein were detected well before the onset of rut in April and coincided with the highest percentage of INSL3-positive cell number/square millimeter of testicular interstitial area. During the winter (December, February), roe deer INSL3 was exclusively detected in a subpopulation of alpha-actin-negative, spindle-shaped peritubular cells. Concordant with the increase in INSL3 production in April and 1 mo after the reported LH peak, a sharp increase in serum testosterone concentrations was observed. High serum testosterone concentrations coincided with the presence of detectable 17alpha-hydroxylase, mRNA and protein, in Leydig cells. Upregulation of INSL3 production in spring appeared to reflect LH-dependent differentiation of Leydig cells. The considerable changes in percentage of INSL3 immunopositive cells within the numerically constant interstitial cell population indicated cyclic seasonal de- and redifferentiation of Leydig cells. A complex functional role of the INSL3/LGR8 ligand-receptor system in the roe deer testis was suggested by the detection of specific hybridization signals for roe deer Lgr8 transcripts in Sertoli cells of the roe deer testis.
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