Male factor infertility is a problem in today’s society but many underlying causes are still unknown. The generation of a conditional Sertoli cell (SC)-specific connexin 43 (Cx43) knockout mouse line (SCCx43KO) has provided a translational model. Expression of the gap junction protein Cx43 between adjacent SCs as well as between SCs and germ cells (GCs) is known to be essential for the initiation and maintenance of spermatogenesis in different species and men. Adult SCCx43KO males show altered spermatogenesis and are infertile. Thus, the present study aims to identify molecular mechanisms leading to testicular alterations in prepubertal SCCx43KO mice. Transcriptome analysis of 8-, 10- and 12-day-old mice was performed by next-generation sequencing (NGS). Additionally, candidate genes were examined by qRT-PCR and immunohistochemistry. NGS revealed many significantly differentially expressed genes in the SCCx43KO mice. For example, GC-specific genes were mostly downregulated and found to be involved in meiosis and spermatogonial differentiation (e.g., Dmrtb1, Sohlh1). In contrast, SC-specific genes implicated in SC maturation and proliferation were mostly upregulated (e.g., Amh, Fshr). In conclusion, Cx43 in SCs appears to be required for normal progression of the first wave of spermatogenesis, especially for the mitosis-meiosis switch, and also for the regulation of prepubertal SC maturation.

Testicular Connexin43 (Cx43) connects adjacent Sertoli cells (SC) and SC to germ cells (GC) in the seminiferous epithelium and plays a crucial role in spermatogenesis. However, the distinction whether this results from impaired inter-SC communication or between GC and SC is not possible, so far. Thus, the question arises, whether a GC-specific Cx43 KO has similar effects on spermatogenesis as it is general or SC-specific KO. Using the Cre/loxP recombinase system, two conditional KO mouse lines lacking Cx43 in premeiotic (pGCCx43KO) or meiotic GC (mGCCx43KO) were generated. It was demonstrated by qRT-PCR that Cx43 mRNA was significantly decreased in adult pGCCx43KO mice, while it was also reduced in mGCCx43KO mice, yet not statistically significant. Body and testis weights, testicular histology, tubular diameter, numbers of intratubular cells and Cx43 protein synthesis and localization did not show any significant differences in semi-quantitative Western blot analysis and immunohistochemistry comparing adult male KO and WT mice of both mouse lines. Male KO mice were fertile. These results indicate that Cx43 in spermatogonia/spermatids does not seem to be essential for successful termination of spermatogenesis and fertility as it is known for Cx43 in somatic SC, but SC-GC communication might rather occur via heterotypic GJ channels.

The aim of this study was to test whether a single testicular needle biopsy could provide histological results comparable to en bloc resection histology and whether one biopsy was sufficient to reflect the histology of an entire pair of testicles. Two methods of sample collection were tested on 32 bull calves aged five to eight months to compare histological parameters of needle biopsy with those of en bloc resection samples. One testicular needle biopsy of the right and three en bloc samples of both testicles were collected and compared for the number of tubular cross sections, tubules with elongated spermatids (ES), outer/inner diameter of tubules, thickness of tubular wall, and number of Sertoli cells (SC). Additionally, animal data were considered. No significant differences were found between the left and right testis or among the individual locations of en bloc samples. However, histologically significant differences (Bonferroni-adjusted significance level: p < 0.05/6 = 0.0083) were found between the needle biopsy and en bloc resection regarding the tubular cross sections per visual field (p < 0.05), the outer (p = 0.01) and inner diameter and the thickness of the tubular wall (both p < 0.01). In the SOX9 immunohistochemical staining, no significant differences (p > 0.05) could be observed for SC numbers between needle biopsy and en bloc samples. In conclusion, results of testicular needle biopsy do not have the same validity as the en bloc resection histology. Furthermore, one biopsy is insufficient to reflect the histology of the entire testicular pair.