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


Journal

Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288

Informations de publication

Date de publication:
28 07 2022
Historique:
received: 22 12 2021
accepted: 18 07 2022
entrez: 28 7 2022
pubmed: 29 7 2022
medline: 2 8 2022
Statut: epublish

Résumé

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.

Identifiants

pubmed: 35902708
doi: 10.1038/s41598-022-16919-4
pii: 10.1038/s41598-022-16919-4
pmc: PMC9334284
doi:

Substances chimiques

Connexin 43 0

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

12898

Informations de copyright

© 2022. The Author(s).

Références

Russel, L. D., Ren, H. P., Sinha Hikim, I., Schulze, W. & Sinha Hikim, A. P. A comparative study in twelve mammalian species of volume densities, volumes, and numerical densities of selected testis components, emphasizing those related to the Sertoli cell. Am. J. Anat. 188, 21–30 (1990).
Russell, L. D. Form, dimensions and cytology of mammalian Sertoli cells in The Sertoli Cell, (eds. Russell, L. D. & Griswold, M. D.) 1–38 (Cache River Press, 1993).
Hess, R. A. & França, L. R. Structure of the Sertoli Cell in Sertoli Cell Biology, (eds. Skinner, M. K. & Griswold, M. D.) 19–40 (Elsevier Academic Press, 2005).
Aumüller, G., Schulze, C. & Viebahn, C. Intermediate filaments in Sertoli cells. Microsc. Res. Tech. 20, 50–72 (1992).
pubmed: 1377059 doi: 10.1002/jemt.1070200107
Suarez-Quian, C. A. & Dym, M. Characterization of Sertoli cell perinuclear filaments. Microsc. Res. Tech. 20, 219–231 (1992).
pubmed: 1543876 doi: 10.1002/jemt.1070200302
Flickinger, C. J. The postnatal development of the Sertoli cells of the mouse. Z. Zellforsch. Mikrosk. Anat. 78, 92–113 (1967).
pubmed: 5596658 doi: 10.1007/BF00344405
Gondos, B. & Berndston, W. E. Postnatal and pubertal development in The Sertoli Cell, (eds. Russell, L. D. & Griswold, M. D.) 115–154 (Cache River Press, 1993).
Bruzzone, R., White, T. W. & Paul, D. L. Connections with connexins: the molecular basis of direct intercellular signalling. Eur. J. Biochem. 238, 1–27 (1996).
pubmed: 8665925 doi: 10.1111/j.1432-1033.1996.0001q.x
Kumar, N. M. & Gilula, N. B. The gap junction communication channel. Cell 84, 381–388 (1996).
pubmed: 8608591 doi: 10.1016/S0092-8674(00)81282-9
Batias, C. et al. Modified expression of testicular gap-junction connexin 43 during normal spermatogenic cycle and in altered spermatogenesis. Cell Tissue Res. 298, 113–121 (1999).
pubmed: 10555545 doi: 10.1007/s004419900076
Pelletier, R. M. The distribution of connexin 43 is associated with the germ cell differentiation and with the modulation of the Sertoli cell junctional barrier in continual (guinea pig) and seasonal breeders’ (mink) testes. J. Androl. 16, 400–409 (1995).
pubmed: 8575979
Decrouy, X., Gasc, J. M., Pointis, G. & Segretain, D. Functional characterization of Cx43 based gap junctions during spermatogenesis. J. Cell Physiol. 200, 146–154 (2004).
pubmed: 15137067 doi: 10.1002/jcp.10473
Steger, K., Tetens, F. & Bergmann, M. Expression of connexin 43 in human testis. Histochem. Cell Biol. 112, 215–220 (1999).
pubmed: 10502068 doi: 10.1007/s004180050409
Sharpe, R. M., McKinnell, C., Kivlin, C. & Fisher, J. S. Proliferation and functional maturation of Sertoli cells, and their relevance to disorders of testis function in adulthood. Reproduction 125, 769–784 (2003).
pubmed: 12773099 doi: 10.1530/rep.0.1250769
Bellvé, A. R., et al. Spermatogenic cells of the prepubertal mouse. Isolation and morphological characterization. J. Cell Biol. 74, 68–85 (1977).
Kluin, P. M., Kramer, M. F. & de Rooij, D. G. Proliferation of spermatogonia and Sertoli cells in maturing mice. Anat. Embryol. 169, 73–78 (1984).
doi: 10.1007/BF00300588
Vergouwen, R. P., Jacobs, S. G., Huiskamp, R., Davids, J. A. & de Rooj, D. G. Proliferative activity of gonocytes, Sertoli cells and interstitial cells during testicular development in mice. J. Reprod. Fertil. 93, 233–243 (1991).
pubmed: 1920294 doi: 10.1530/jrf.0.0930233
Sridharan, S. et al. Proliferation of adult sertoli cells following conditional knockout of the Gap junctional protein GJA1 (connexin 43) in mice. Biol. Reprod. 76, 804–812 (2007).
pubmed: 17229929 doi: 10.1095/biolreprod.106.059212
Weider, K. Untersuchungen zum Differenzierungszustand von Sertoli Zellen in Mäusen mit Sertoli zellspezifischen Knockout des Connexin 43-Gens (SCCx43KO). VVB Laufersweiler. (2011)
Weider, K. et al. Altered differentiation and clustering of Sertoli cells in transgenic mice showing a Sertoli cell specific knockout of the connexin 43 gene. Differentiation 82, 38–49 (2011).
pubmed: 21489682 doi: 10.1016/j.diff.2011.03.001
Brehm, R. et al. A sertoli cell-specific knockout of connexin43 prevents initiation of spermatogenesis. Am. J. Pathol. 171, 19–31 (2007).
pubmed: 17591950 pmcid: 1941593 doi: 10.2353/ajpath.2007.061171
Rode, K. et al. Loss of connexin 43 in Sertoli cells provokes postnatal spermatogonial arrest, reduced germ cell numbers and impaired spermatogenesis. Reprod. Biol. 18, 456–466 (2018).
pubmed: 30243528 doi: 10.1016/j.repbio.2018.08.001
Carette, D. et al. Major involvement of connexin 43 in seminiferous epithelial junction dynamics and male fertility. Dev. Biol. 346, 54–67 (2010).
pubmed: 20655897 doi: 10.1016/j.ydbio.2010.07.014
Hollenbach, J., et al. Loss of connexin43 in murine Sertoli cells and its effects on blood-testis barrier formation and dynamics. PLoS One. 13, e0198100. https://doi.org/10.1371/journal.pone.0198100 (2018).
Gerber, J., Heinrich, J. & Brehm, R. Blood-testis barrier and Sertoli cell function: lessons from SCCx43KO mice. Reproduction. 151, R15–27. https://doi.org/10.1530/REP-15-0366 (2016).
Wong, V. & Russell, L. D. Three-dimensional reconstruction of a rat stage V Sertoli cell: I. Methods, basic configuration, and dimensions. Am. J. Anat. 167, 143–161 (1983).
Starborg, T. & Kadler, K. E. Serial block face-scanning electron microscopy: A tool for studying embryonic development at the cell-matrix interface. Birth Defects Res. C. Embryo Today. 105, 9–18 (2015).
pubmed: 25820245 doi: 10.1002/bdrc.21087
Rizzo, N. W., Duncan, K. E., Bourett, T. M. & Howard, R. J. Backscattered electron SEM imaging of resin sections from plant specimens: observation of histological to subcellular structure and CLEM. J. Microsc. 263, 142–147 (2016).
pubmed: 26708578 doi: 10.1111/jmi.12373
Handel, M. A. & Eppig, J. J. Sertoli cell differentiation in the testes of mice genetically deficient in germ cells. Biol. Reprod. 20, 1031–1038 (1979).
pubmed: 476239 doi: 10.1095/biolreprod20.5.1031
Young, G. P. et al. Sertoli cell-only syndrome produced by cold testicular ischemia. Endocrinology 122, 1074–1082 (1988).
pubmed: 3125037 doi: 10.1210/endo-122-3-1074
Wang, Z. et al. Sertoli cell-only phenotype and scRNA-seq define PRAMEF12 as a factor essential for spermatogenesis in mice. Nat. Commun. 10, 1–18 (2019).
Yamashita, Y., Suzuki, C., Uchiyama, Y. & Nagata, S. Infertility caused by inefficient apoptotic germ cell clearance in Xkr8-deficient male mice. Mol. Cell Biol. 40, e00402–19. https://doi.org/10.1128/MCB.00402-19 (2020).
Chung, K. W. A morphological and histochemical study of sertoli cells in normal and XX sex-reversed mice. Am. J. Anat. 139, 369–387 (1974).
doi: 10.1002/aja.1001390306
Kerr, J. B., Rich, K. A. & de Kretser, D. M. Effects of experimental cryptorchidism on the ultrastructure and function of the sertoli cell and peritubular tissue of the rat testis. Biol. Reprod. 21, 823–838 (1979).
pubmed: 43167 doi: 10.1095/biolreprod21.4.823
Dirami, G., Ravindranath, N., Kleinmann, H. K. & Dym, M. Evidence that basement membrane prevents apoptosis of Sertoli cells in vitro in the absence of known regulators of Sertoli cell function. Endocrinology 136, 4439–4447 (1995).
pubmed: 7664664 doi: 10.1210/endo.136.10.7664664
Wright, W. W., Fiore, C. & Zirkin, B. R. The effect of aging on the seminiferous epithelium of the brown Norway rat. J. Androl. 14, 110–117 (1993).
pubmed: 8514616
Millner, A. & Atilla-Gokcumen, G. E. Lipid players of cellular senescence. Metabolites 10, 339 (2020).
pmcid: 7570155 doi: 10.3390/metabo10090339
Nakanishi, Y. & Shiratsuchi, A. Phagocytic removal of apoptotic spermatogenic cells by Sertoli cells: Mechanisms and consequences. Biol. Pharm. Bull. 27, 13–16 (2004).
pubmed: 14709891 doi: 10.1248/bpb.27.13
Chemes, H. The phagocytic function of Sertoli cells: A morphological, biochemical, and endocrinological study of lysosomes and acid phosphatase localization in the rat testis. Endocrinology 119, 1673–1681 (1986).
pubmed: 3757907 doi: 10.1210/endo-119-4-1673
Kerr, J. B. & de Kretser, D. M. Cyclic variations in Sertoli cell lipid content throughout the spermatogenic cycle in the rat. Reprod. Fertil. 43, 1–8 (1975).
doi: 10.1530/jrf.0.0430001
Kerr, J. B., Mayberry, R. A. & Irby, D. C. Morphometric studies on lipid inclusions in Sertoli cells during the spermatogenic cycle in the rat. Cell Tissue Res. 236, 699–709 (1984).
pubmed: 6088048 doi: 10.1007/BF00217241
Xiong, W., Wang, H., Wu, H., Chen, Y. & Han, D. Apoptotic spermatogenic cells can be energy sources for Sertoli cells. Reproduction 137, 469–479 (2009).
pubmed: 19074501 doi: 10.1530/REP-08-0343
Nistal, M., Jimenez, F. & Paniagua, R. Sertoli cell types in the Sertoli-cell-only syndrome: Relationships between Sertoli cell morphology and aetiology. Histopathology 16, 173–180 (1990).
pubmed: 2182507 doi: 10.1111/j.1365-2559.1990.tb01086.x
Noelke, J. 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).
pubmed: 25693895 doi: 10.1007/s00441-015-2126-7
Tarulli, G. A., Stanton, P. G., Lerchl, A. & Meachem, S. J. Adult sertoli cells are not terminally differentiated in the Djungarian hamster: Effect of FSH on proliferation and junction protein organization. Biol. Reprod. 76, 798–806 (2006).
doi: 10.1095/biolreprod.105.050450
Ahmed, E. A. et al. Proliferative activity in vitro and DNA repair indicate that adult mouse and human Sertoli cells are not terminally differentiated, quiescent cells. Biol. Reprod. 80, 1084–1091 (2009).
pubmed: 19164176 doi: 10.1095/biolreprod.108.071662
Solari, A. J. & Fritz, I. B. The ultrastructure of immature Sertoli cells. Maturation-like changes during culture and the maintenance of mitotic potentiality. Biol. Reprod. 18, 329–345 (1978)
Kulibin, A. Y. & Malolina, E. A. Only a small population of adult Sertoli cells actively proliferates in culture. Reproduction 152, 271–281 (2016).
pubmed: 27512121 doi: 10.1530/REP-16-0013
Gilleron, J. et al. A potential novel mechanism involving connexin 43 gap junction for control of sertoli cell proliferation by thyroid hormones. J. Cell Physiol. 209, 153–161 (2006).
pubmed: 16823880 doi: 10.1002/jcp.20716
Gilleron, J., Carette, D., Durand, P., Pointis, G. & Segretain, D. Connexin 43 a potential regulator of cell proliferation and apoptosis within the seminiferous epithelium. Int. J. Biochem. Cell Biol. 41, 1381–1390 (2009).
pubmed: 19136074 doi: 10.1016/j.biocel.2008.12.008
Gaytan, F. & Aguilar, E. Quantitative analysis of Sertoli cells in neonatally oestrogen-treated rats. J. Reprod. Fertil. 79, 589–598 (1987).
pubmed: 3572890 doi: 10.1530/jrf.0.0790589
Chemes, H. E., Dym, M. & Raj, H. G. Hormonal regulation of Sertoli cell differentiation. Biol. Reprod. 21, 251–262 (1979).
pubmed: 486642 doi: 10.1095/biolreprod21.1.251
McLain, R. F. & Weinstein, J. N. Nuclear clefting in dorsal root ganglion neurons: A response to whole body vibration. J. Comp. Neurol. 322, 538–547 (1992).
pubmed: 1401248 doi: 10.1002/cne.903220408
Graveland, G. A. & DiFiglia, M. The frequency and distribution of medium-sized neurons with indented nuclei in the primate and rodent neostriatum. Brain Res. 327, 307–311 (1985).
pubmed: 3986508 doi: 10.1016/0006-8993(85)91524-0
Buschmann, M. T., Geoffroy, J. S. & LaVelle, A. Nuclear envelope invaginations in hamster pyramidal cells during development and aging. Neurobiol. Aging. 2, 27–32 (1981).
pubmed: 7266739 doi: 10.1016/0197-4580(81)90055-5
Burkl, W. & Thiel-Bartosch, E. Elektronenmikroskopische Untersuchungen über die Granulosa atresierender Tertiärfollikel bei der Ratte. Arch. Gynakol. 204, 238–250 (1967).
pubmed: 4171694 doi: 10.1007/BF00669405
Mestwerdt, W., Müller, O. & Brandau, H. Light and electronmicroscopic examinations on granulosa and theca of preovulatory and freshly ruptured follicles of human ovaries (author’s transl). Arch. Gynakol. 222, 115–136 (1977).
pubmed: 577125 doi: 10.1007/BF00667196
Paulini, F., Silva, R. C., Rôlo, J. L. & Lucci, C. M. Ultrastructural changes in oocytes during folliculogenesis in domestic mammals. J. Ovarian Res. 7, 102 (2014).
pubmed: 25358389 pmcid: 4224757 doi: 10.1186/s13048-014-0102-6
Rothbarth, K. et al. Intracellular location and nuclear targeting of the Spi-1, Spi-2 and Spi-3 gene-derived serine protease inhibitors in non-secretory cells. Eur. J. Cell Biol. 80, 341–348 (2001).
pubmed: 11432724 doi: 10.1078/0171-9335-00170
Bourgeois, C. A., Hemon, D. & Bouteille, M. Structural relationship between the nucleolus and the nuclear envelope. J. Ultrastruct. Res. 68, 328–340 (1979).
pubmed: 490761 doi: 10.1016/S0022-5320(79)90165-5
Fricker, M., Hollinshead, M., White, N. & Vaux, D. Interphase nuclei of many mammalian cell types contain deep, dynamic, tubular membrane-bound invaginations of the nuclear envelope. J. Cell Biol. 36, 531–544 (1997).
doi: 10.1083/jcb.136.3.531
Hirano, Y. et al. Lamin B receptor recognizes specific modifications of histone H4 in heterochromatin formation. J. Biol. Chem. 287, 42654–42663 (2012).
pubmed: 23100253 pmcid: 3522266 doi: 10.1074/jbc.M112.397950
Solovei, I. et al. LBR and lamin A/C sequentially tether peripheral heterochromatin and inversely regulate differentiation. Cell 152, 584–598 (2013).
pubmed: 23374351 doi: 10.1016/j.cell.2013.01.009
Belevich, I., Joensuu, M., Kumar, D., Vihinen, H. & Jokitalo, E. Microscopy image browser: A platform for segmentation and analysis of multidimensional datasets. PLoS Biol. 14, 2340. https://doi.org/10.1371/journal.pbio.1002340 (2016).
doi: 10.1371/journal.pbio.1002340
Romeis, B. Darstellung von paraplasmatischen Substanzen in Romeis Mikroskopische Technik, (ed. Böck, P.) 375–408 (Urban und Schwarzenberg, 1989)
Riedelsheimer, B. & Büchl-Zimmermann, S. Färbungen in Romeis Mikroskopische Technik, (eds. Mulisch, M. & Welsch, U.) 171–282 (Springer Spektrum, 2015)
Liu, M. et al. Transient scrotal hyperthermia induces lipid droplet accumulation and reveals a different ADFP expression pattern between the testes and liver in mice. PLoS ONE 7, 5694. https://doi.org/10.1371/journal.pone.0045694 (2012).
doi: 10.1371/journal.pone.0045694

Auteurs

Sarah Staggenborg (S)

Institute of Anatomy, University of Veterinary Medicine Hannover, Foundation, Bischofsholer Damm 15, 30173, Hannover, Germany. sarah.staggenborg@tiho-hannover.de.

Rüdiger Koch (R)

Institute of Anatomy, University of Veterinary Medicine Hannover, Foundation, Bischofsholer Damm 15, 30173, Hannover, Germany.

Kristina Rode (K)

Institute of Anatomy, University of Veterinary Medicine Hannover, Foundation, Bischofsholer Damm 15, 30173, Hannover, Germany.

Hanna Hüneke (H)

Institute of Anatomy, University of Veterinary Medicine Hannover, Foundation, Bischofsholer Damm 15, 30173, Hannover, Germany.

Louiza Tiedje (L)

Institute of Anatomy, University of Veterinary Medicine Hannover, Foundation, Bischofsholer Damm 15, 30173, Hannover, Germany.

Gudrun Wirth (G)

Institute of Anatomy, University of Veterinary Medicine Hannover, Foundation, Bischofsholer Damm 15, 30173, Hannover, Germany.

Marion Langeheine (M)

Institute of Anatomy, University of Veterinary Medicine Hannover, Foundation, Bischofsholer Damm 15, 30173, Hannover, Germany.

Ines Blume (I)

Institute of Anatomy, University of Veterinary Medicine Hannover, Foundation, Bischofsholer Damm 15, 30173, Hannover, Germany.

Kerstin Rohn (K)

Institute of Pathology, University of Veterinary Medicine Hannover, Foundation, Hannover, Germany.

Christoph Wrede (C)

Institute of Functional and Applied Anatomy, Research Core Unit Electron Microscopy, Hannover Medical School, Hannover, Germany.

Christiane Pfarrer (C)

Institute of Anatomy, University of Veterinary Medicine Hannover, Foundation, Bischofsholer Damm 15, 30173, Hannover, Germany.

Ralph Brehm (R)

Institute of Anatomy, University of Veterinary Medicine Hannover, Foundation, Bischofsholer Damm 15, 30173, Hannover, Germany.

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