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A Sertoli cell-specific connexin43 knockout leads to altered interstitial connexin expression and increased Leydig cell numbers

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Abstract

The Sertoli cell (SC)-specific knockout (KO) of connexin43 (Cx43) results in spermatogenic arrest at the level of spermatogonia and/or SC-only syndrome. Histology of the interstitial compartment suggests Leydig cell (LC) hyperplasia. Our aim has been to investigate possible effects of the SC-specific KO of Cx43 (SCCx43KO) on interstitial LC. We therefore counted LC via the optical dissector method (per microliter of testicular tissue and per testis) and found LC to be significantly increased in SCCx43KO−/− compared with wild-type mice. Semiquantitative western blot together with Cx43 and 3β-hydroxysteroid dehydrogenase immunohistochemistry showed that Cx43 protein was significantly reduced and barely detectable in LC in adult SCCx43KO−/− mice. This reduction of Cx43 protein was accompanied by a reduction of Cx43 mRNA as analyzed by laser-assisted microdissection of interstitial cells and subsequent quantitative real-time polymerase chain reaction (PCR). Interestingly, Cx45, another recently detected connexin in LC, was also downregulated. Preliminary qualitative data of LC differentiation markers (Thb2, Hsd3b6) and a steroidogenic marker (Hsd17b3) obtained by reverse transcription plus PCR revealed no obvious differences. Thus, the loss of Cx43 in SC also provokes the downregulation of connexins in interstitial LC at the transcriptional and translational levels. Moreover, SCCx43KO leads to alterations in LC numbers. Despite these alterations, steroidogenesis seems not to be impaired. Further studies, including ultrastructural analysis of the tissue as well as quantitative examination of additional LC markers and testosterone, and functional in vitro experiments, should provide more information about LC differentiation and function in SCCx43KO−/− mice.

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Acknowledgements

We thank Dr. N. Hambruch (Institute of Anatomy, University of Veterinary Medicine Hannover, Foundation, Germany) for her advice and technical assistance with the laser microdissection approach and statistical analysis. We also thank Prof. J.I. Mason (Centre for Reproductive Biology, University of Edinburgh, Scotland) for providing the 3β-HSD antibody. We also gratefully acknowledge the technical support of Jutta Salzig, Centre of Reproductive Medicine and Andrology, Muenster.

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Authors and Affiliations

  1. Institute of Anatomy, University of Veterinary Medicine Hannover, Foundation, 30173, Hannover, Germany

    Joanna Noelke, Jonathan Gerber, Marion Gaehle & Ralph Brehm

  2. Centre of Reproductive Medicine and Andrology, University Muenster, 48149, Muenster, Germany

    Joachim Wistuba & Oliver S. Damm

  3. Institute of Veterinary Anatomy, Histology and Embryology, University of Giessen, 35392, Giessen, Germany

    Daniela Fietz

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  1. Joanna Noelke
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  2. Joachim Wistuba
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  5. Jonathan Gerber
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  6. Marion Gaehle
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  7. Ralph Brehm
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Correspondence to Ralph Brehm.

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This work was financially supported by the Kogge Foundation, Giessen, Germany.

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Noelke, J., Wistuba, J., Damm, O.S. et al. A Sertoli cell-specific connexin43 knockout leads to altered interstitial connexin expression and increased Leydig cell numbers. Cell Tissue Res 361, 633–644 (2015). https://doi.org/10.1007/s00441-015-2126-7

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