Cycle Of Rat Seminiferous Epithelium Research Articles

The Effects of Triiodothyronine on Rat Testis: A Morphometric and Immunohistochemical Study

The aim of present study was to investigate the effects of 3,3',5-triiodothyronine (T(3)) on rat testis both morphometrically and immunohistochemically with determining of insulin-like growth factor I (IGF-I) expression. Adult male Wistar-albino rats used in the study were divided into two groups; control and T(3)-treated groups. After T(3) treatment there was observed to be a decrease in testicular weights, diameters of seminiferous tubules and the number of sertoli cells, and an increase in the number of leydig cells (P<0.05). Some of the seminiferous tubule lumens of T(3) administrated rats had cellular debris. IGF-I was localized in sertoli cells, late spermatids and leydig cells of all groups. IGF-I immunoreactivity in T(3) treated rats was higher than in controls in all stages of the cycle of rat seminiferous epithelium, but the staining intensity of leydig cells were similar in both groups. In conclusion, the present results suggest that T(3) may modulate the testicular function by affecting IGF-I activity at the gonadal level.

Expression of Mitochondrial Heat Shock Protein 60 in Distinct Cell Types and Defined Stages of Rat Seminiferous Epithelium1

Changes in the level of the gene transcript of heat shock protein (hsp)60, a mitochondrial chaperonin, during the cycle of rat seminiferous epithelium and its cellular localization were studied. The seminiferous epithelium showed a cell type-specific expression of hsp60. Immunostaining of adult rat testis revealed localization in Sertoli and Leydig cells. In germ cells, mitochondria of spermatogonia and early primary spermatocytes were immunoreactive for hsp60. Mitochondria of all other germ cell types were completely negative for hsp60. Stage-specific expression of hsp60 was determined from pooled segments of stage-specific microdissected tubules by a combination of Western blotting and polymerase chain reaction (PCR). High concentrations of hsp60 were found in stages I-V and IX-XIV, and low levels were detected in the other stages, i.e., VI-VIII. In stages with high hsp60 expression, spermatogonia divide mitotically, whereas in stages lacking mitosis, the hsp60 level was much weaker. In seminiferous epithelium, two different types of mitochondria are present. Therefore, immunoelectron microscopy was used to differentiate these two morphologically distinct types of mitochondria. The crista type of mitochondria (e.g., in Sertoli cells and spermatogonia) reacted with the antibody against hsp60, whereas hsp60 was negative in so-called "condensed"-type mitochondria found in midpachytene spermatocytes and more advanced germ cells. It could be shown for the first time that expression of the hsp60 gene is regulated during the cycle of the seminiferous epithelium. The results indicate that the gene product is primarily needed during the initial steps of spermatogenesis in which most of the cell divisions occur, while its expression during the differentiation of spermatids and sperm is obviously not necessary. The presence of hsp60 in stages with mitotic activity suggests a very active mitochondrial protein import and protein assembly machinery that generates further mitochondria for the dividing cells.

Expression of a testis-specific hsp70 gene-related RNA in defined stages of rat seminiferous epithelium.

Changes in the level of a testis-specific hsp70 gene-related transcript (hst70 RNA) and its cellular localization during the cycle of rat seminiferous epithelium have been investigated. Segments of seminiferous tubules at defined stages of the cycle were isolated in living condition by transillumination-assisted microdissection and the exact stages identified by phase-contrast microscopy of live cell squashes. The levels of the hst70 RNA were determined by Northern and slot blotting of whole cell lysates. High levels were found in stages XII-XIV and I to early VII of the cycle, and low levels were found in other stages, i.e., late VII (VIId) through VIII-XI of the cycle. The in situ hybridization revealed that the hst70 gene was activated in late pachytene primary spermatocytes during stage XII of the cycle, and that mRNA was then present in cells during differentiation through diakinesis, meiotic divisions, and early spermiogenesis (steps 1 through early 7). The activation of the gene coding for hst70 RNA shortly before meiotic divisions may indicate that the gene product is needed either during differentiation of late spermatocytes into spermatids or later during spermiogenesis, and that the mRNA may be stored in early spermatids.