Effects of a Sertoli cell-specific knockout of Connexin43 on maturation and proliferation of postnatal Sertoli cells.

Adult male Sertoli cell-specific Connexin43 knockout mice (SCCx43KO) exhibit higher Sertoli cell (SC) numbers per seminiferous tubule compared to their wild type (WT) littermates. Thus, deletion of this testicular gap junction protein seems to affect the proliferative potential and differentiation of "younger" SC. Although SC have so far mostly been characterised as postmitotic cells that cease to divide and become an adult, terminally differentiated cell population at around puberty, there is rising evidence that there exist exceptions from this for a very long time accepted paradigm. Aim of this study was to investigate postnatal SC development and to figure out underlying causes for observed higher SC numbers in adult KO mice. Therefore, the amount of SC mitotic figures was compared, resulting in slightly more and prolonged detection of SC mitotic figures in KO mice compared to WT. SC counting per tubular cross section revealed significantly different time curves, and comparing proliferation rates using Bromodesoxyuridine and Sox9 showed higher proliferation rates in 8-day old KO mice. SC proliferation was further investigated by Ki67 immunohistochemistry. SC in KO mice displayed a delayed initiation of cell-cycle-inhibitor p27Kip1 synthesis and prolonged synthesis of the phosphorylated tumour suppressor pRb and proliferation marker Ki67. Thus, the higher SC numbers in adult male SCCx43KO mice may arise due to two different reasons: Firstly, in prepubertal KO mice, the proliferation rate of SC was higher. Secondly, there were differences in their ability to cease proliferation as shown by the delayed initiation of p27Kip1 synthesis and the prolonged production of phosphorylated pRb and Ki67. Immunohistochemical results indicating a prolonged period of SC proliferation in SCCx43KO were confirmed by detection of proliferating SC in 17-days-old KO mice. In conclusion, deletion of the testicular gap junction protein Cx43 might prevent normal SC maturation and might even alter also the proliferation potential of adult SC.

Connexin43 represents an important regulator for Sertoli cell morphology, Sertoli cell nuclear ultrastructure, and Sertoli cell maturation.

The Sertoli cell (SC)-specific knockout (KO) of connexin43 (Cx43) was shown to be an effector of multiple histological changes in tubular morphology, resulting in germ cell loss through to a Sertoli-cell-only (SCO) phenotype and vacuolated seminiferous tubules containing SC-clusters. Our present study focused on the effects of Cx43 loss on SC ultrastructure. Using serial block-face scanning electron microscopy (SBF-SEM), we could confirm previous results. Ultrastructural analysis of Sertoli cell nuclei (SCN) revealed that these appear in clusters with a phenotype resembling immature/proliferating SCs in KO mice. Surprisingly, SCs of fertile wild type (WT) mice contained SCN with a predominantly smooth surface instead of deep indentations of the nuclear envelope, suggesting that these indentations do not correlate with germ cell support or spermatogenesis. SBF-SEM facilitated the precise examination of clustered SCs. Even if the exact maturation state of mutant SCs remained unclear, our study could detect indications of cellular senescence as well as immaturity, emphasising that Cx43 affects SC maturation. Moreover, Sudan III staining and transmission electron microscopy (TEM) demonstrated an altered lipid metabolism in SCs of Cx43 deficient mice.

Precocious puberty in male wild boars: a possible explanation for the dramatic population increase in Germany and Europe.

BACKGROUND: The wild boar population in Europe is steadily growing, one of the reasons for this increase probably being the high reproductive potential of this large mammal. Population management is important to stabilise wild boar numbers and a great deal of attention is focusing on the reasons, which might contribute to the high reproductive rates. Understanding the timing of puberty attainment provides information required for proper management practices. Knowledge of the earliest expected time of sexual maturation in male wild boars is limited, research being mostly focused on females. Previous hunting references indicate that sexual maturity in males occurs in the second year after birth. In contrast, male domestic pigs become sexually mature from about seven months of age. Thus, aims of this study were to investigate (1) whether there is a physiological ability for reproduction also in male wild boars of a younger age and (2) whether the body weight of wild boar males has a more important role than age in driving the maturation of the testis. METHODS: Male wild boar individuals were sampled during hunting drives in the eastern part of Lower Saxony in Germany. Testes with epididymides from 74 males were collected and prepared for histological examination and immunohistochemistry. The reproductive status could be ascertained based on development/occurrence of different germ cell populations using histology and based on the immunohistochemical detection of the anti-Müllerian hormone and androgen receptor. RESULTS: In this study, male wild boars aged nine to ten months already passed puberty and were able to reproduce if they had reached the appropriate body condition of about 29 kg dressed weight. Immunopositivity to the anti-Müllerian hormone in Sertoli cells was evident only in prepubertal animals and decreased with the onset of puberty. No immunoreaction was evident at postpuberty. The androgen receptor was detected in Sertoli cells, peritubular cells and Leydig cells, surprisingly already in Sertoli cells of prepubertal wild boars as well depending on body weight. Moreover, two-thirds of young males aged about ten months were precociously reproductively mature, showing histologically the presence of spermatozoa in testes and epididymides. CONCLUSIONS: As piglets are mostly born in spring, also these young male individuals could target the heat of female wild boars in the winter months, resulting in the observed population increase. Therefore, a reduction in wild boar numbers should also focus on piglets of both sexes.