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Effects of a murine germ cell-specific knockout of Connexin 43 on Connexin expression in testis and fertility

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Abstract

Connexin 43 (Cx 43)—expressed by germ cells (GC), Sertoli cells (SC) and Leydig cells—is one of at least eleven Cx in the murine testis. A general knockout (KO) of Cx 43 in mice results in perinatal death and a SC-specific KO of Cx 43 (SCCx43KO) causes infertility of male mice by preventing the initiation of spermatogenesis. To further elucidate the role of Cx 43 in the testis, a new mouse model with a GC-specific KO of Cx 43 (GCCx43KO) was created by using the Cre/loxP recombination system. A transgenic mouse line expressing the Cre gene under the tissue non-specific alkaline phosphatase promoter and a transgenic floxed Cx 43-LacZ mouse line were mated. The resulting F1-generation was backcrossed with homozygous Cx 43 floxed mice, and offspring was genotyped. Immunohistochemical analysis of testes of different aged homozygous mice revealed normal spermatogenesis and reduced Cx 43 immunoreactions. RT-qPCR and Western blots showed a downregulation of Cx 43 mRNA and protein, and a nearly unchanged mRNA expression of Cx 26, Cx 33 and Cx 45 in pubertal and adult KO mice. Western blots revealed considerable immunoreactive bands for Cx 26 and Cx 45. Male and female homozygous GCCx43KO mice were viable and fertile. Our data suggest, in contrast to inter SC and inter SC–GC cross talk in SCCx43KO mice which depends selectively on Cx 43 expression, that Cx 43 in GC seems not to be essential in GC–SC communication, when other Cx persist to be expressed.

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Acknowledgments

We thank D. Schaefer and his crew at the Animal Facility, University of Marburg, Marburg, Germany, for their professional and noteworthy help with treatment of our transgenic mice, A. Hax (Giessen, Germany) and M. Gähle and S. J. Schultz (both Hannover, Germany) for their skilful technical assistance. We thank Professor K. Willecke (Institute of Genetics, University of Bonn, Germany) for the generous provision of the floxed Cx43-LacZ transgenic mice and finally Professor H. Lomelí (Department of Developmental Genetics and Molecular Physiology, Institute of Biotechnology, National Autonomous University of Mexico, Mexico) and Professor A. Nagy (Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Canada) for the permissive provision of the TNAP-Cre mice. The authors address special thanks to Dr. D. Carette and Professor J. Nagy for their data about Connexin antibodies. This work was supported by the DFG (BR 3365/2-1 and KFO 181/1) and by the AfT (Akademie für Tiergesundheit), Bonn, Germany.

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Authors and Affiliations

  1. Institute for Veterinary Anatomy, Histology and Embryology, Justus Liebig University Giessen, Frankfurter Str. 98, 35392, Giessen, Germany

    Sabine Günther, Daniela Fietz & Martin Bergmann

  2. Department of Anatomy, University of Veterinary Medicine Hannover, Bischofsholer Damm 15, 30173, Hannover, Germany

    Karola Weider & Ralph Brehm

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  1. Sabine Günther
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  2. Daniela Fietz
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  3. Karola Weider
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  4. Martin Bergmann
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  5. Ralph Brehm
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Correspondence to Ralph Brehm.

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Günther, S., Fietz, D., Weider, K. et al. Effects of a murine germ cell-specific knockout of Connexin 43 on Connexin expression in testis and fertility. Transgenic Res 22, 631–641 (2013). https://doi.org/10.1007/s11248-012-9668-1

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  • DOI: https://doi.org/10.1007/s11248-012-9668-1

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