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Phagocytosis by Sertoli Cells: Analysis of Main Phagocytosis Steps by Confocal and Electron Microscopy

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Sertoli Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1748))

Abstract

Sertoli cells were discovered in the seminiferous tubules by Enrico Sertoli in 1865 (Morgagni 7:31–33, 1865). Intense phagocytosis is, in the context of spermatogenesis cycle, morphologically the most noticeable function of Sertoli cells. In this chapter the major principles of phagocytosis machinery and its specificities in the seminiferous tubules will be briefly reviewed, guidelines of analysis of main phagocytosis steps by confocal and transmission electron microscopy will be described, and a simplified method to assess phagocytosis rate in routine experiments will be given.

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References

  1. Jégou B (1993) The Sertoli-germ cell communication network in mammals. Int Rev Cytol 147:25–96

    Article  PubMed  Google Scholar 

  2. Lacy D (1960) Light and electron microscopy, and its use in the study of factors influencing spermatogenesis in the rat. J R Microsc Soc 79:209–225

    Article  CAS  PubMed  Google Scholar 

  3. Maderna P, Godson C (2003) Phagocytosis of apoptotic cells and the resolution of inflammation. Biochim Biophys Acta 1639:141–151

    Article  CAS  PubMed  Google Scholar 

  4. Rabinovitch M (1995) Professional and non-professional phagocytes: an introduction. Trends Cell Biol 5:85–87

    Article  CAS  PubMed  Google Scholar 

  5. Metchnikoff E (1887) Sur la lutte des cellules de l’organisme contre l’invasion des microbes. Ann Inst Pasteur 1:321–345

    Google Scholar 

  6. Nydegger UE, Miescher A, Anner RM et al (1973) Serum and cellular factor involvement in nitroblue tetrazolium (NBT) reduction by human neutrophils. Klin Wochenschr 51:377–382

    Article  CAS  PubMed  Google Scholar 

  7. Albrecht D, Jungi TW (1993) Luminol-enhanced chemiluminescence induced in peripheral blood-derived human phagocytes: obligatory requirement of myeloperoxidase exocytosis by monocytes. J Leukoc Biol 54:300–306

    Article  CAS  PubMed  Google Scholar 

  8. Kampen AH, Tollersrud T, Larsen S et al (2004) Repeatability of flow cytometric and classical measurement of phagocytosis and respiratory burst in bovine polymorphonuclear leucocytes. Vet Immunol Immunopathol 97:105–114

    Article  CAS  PubMed  Google Scholar 

  9. Lemke G, Rothlin CV (2008) Immunobiology of the TAM receptors. Nat Rev Immunol 8(5):327–336

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Irschik EU, Sgonc R, Böck G et al (2004) Retinal pigment epithelial phagocytosis and metabolism differ from those of macrophage. Ophthalmic Res 36:200–210

    Article  Google Scholar 

  11. Shiratsuchi A, Osada Y, Nakanishi Y (2013) Differences in the mode of phagocytosis of bacteria between macrophages and testicular Sertoli cells. Drug Discov Ther 7(2):73–77

    CAS  PubMed  Google Scholar 

  12. Kevany BM, Palczewsky K (2010) Phagocytosis of retinal rod and cone photoreceptors. Physiology 25:8–15

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Leblond CP, Clermont Y (1952) Definition of the stages of the cycle of the seminiferous epithelium in the rat. Ann N Y Acad Sci 55(4):548–573

    Article  CAS  PubMed  Google Scholar 

  14. Oakberg EF (1956) Duration of spermatogenesis in the mouse and timing of stages of the cycle of the seminiferous epithelium. Am J Anat 99(3):507–516

    Article  CAS  PubMed  Google Scholar 

  15. Clermont Y (1963) The cycle of the seminiferous epithelium in man. Am J Anat 112:35–51

    Article  CAS  PubMed  Google Scholar 

  16. Clermont Y (1972) Kinetics of spermatogenesis in mammals: seminiferous epithelium cycle and spermatogonial renewal. Physiol Rev 52(1):198–236

    Article  CAS  PubMed  Google Scholar 

  17. Chemes H (1986) The phagocytic function of Sertoli cells: a morphological, biochemical and endocrinological study of lysosomes and acid phosphatase localization in the rat testis. Endocrinology 119(4):1673–1681

    Article  CAS  PubMed  Google Scholar 

  18. Blanco-Rodriguez J, Martinez-Garcia C (1999) Apoptosis is physiologically restricted to a specialized cytoplasmic compartment in rat spermatids. Biol Reprod 61:1541–1547

    Article  CAS  PubMed  Google Scholar 

  19. Russell LD, Ettlin RA, Hikim APS, Clegg ED (1990) Histological and histopathological evaluation of the testis. Cache River Press, St. Louis

    Google Scholar 

  20. Pineau C, Le Magueresse B, Courtens JL, Jégou B (1991) Study in vitro of the phagocytic function of Sertoli cells in the rat. Cell Tissue Res 264(3):589–598

    Article  CAS  PubMed  Google Scholar 

  21. Grandjean V, Sage J, Ranc F et al (1997) Stage-specific signals in germ line differentiation: control of Sertoli cell phagocytic activity by spermatogenic cells. Dev Biol 184:165–174

    Article  CAS  PubMed  Google Scholar 

  22. Gillot I, Jehi-Petri C, Gounon P et al (2005) Germ cells and fatty acids induce translocation of CD36 scavenger receptor to the plasma membrane of Sertoli cells. J Cell Sci 118:3027–3035

    Article  CAS  PubMed  Google Scholar 

  23. Yamada H, Ohashi E, Abe T et al (2007) Amphiphysin 1 is important for actin polymerization during phagocytosis. Mol Biol Cell 18(11):4669–4680

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Otsuka A, Abe T, Watanabe M et al (2009) Dynamin 2 is required for actin assembly in phagocytosis in Sertoli cells. Biochem Biophys Res Commun 378:478–482

    Article  CAS  PubMed  Google Scholar 

  25. Hu W, Wang H, Xiong W et al (2006) Evaluation on the phagocytosis of apoptotic spermatogenic cells by Sertoli cells in vitro through detecting lipid droplet formation by Oil Red O staining. Reproduction 132:485–492

    Article  Google Scholar 

  26. Eliott MR, Zheng S, Park D et al (2010) Unexpected requirement for ELMO1 in clearance of apoptotic germ cells in vivo. Nature 467:333–339

    Article  Google Scholar 

  27. Yefimova MG, Sow A, Fontaine I et al (2008) Dimeric transferrin inhibits phagocytosis of residual bodies by testicular rat Sertoli cells. Biol Reprod 78(4):697–704

    Article  CAS  PubMed  Google Scholar 

  28. Meier T, Arni S, Malarkannan S et al (1992) Immunodetection of biotinylated lymphocyte-surface proteins by enhanced chemiluminescence: a nonradioactive method for cell-surface protein analysis. Anal Biochem 204:220–226

    Article  CAS  PubMed  Google Scholar 

  29. Shiratsuchi A, Umeda M, Ohba Y, Nakanishi Y (1997) Recognition of phosphatidylserine on the surface of apoptotic spermatogenic cells and subsequent phagocytosis by Sertoli cells of the rat. J Biol Chem 272(4):2354–2358

    Article  CAS  PubMed  Google Scholar 

  30. Yefimova MG, Messaddeq N, Harnois T et al (2013) A chimerical phagocytosis model reveals the recruitment by Sertoli cells of autophagy for the degradation of ingested illegitimate substrates. Autophagy 9(5):653–666

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Shiratsuchi A, Kawasaki Y, Ikemoto M et al (1999) Role of class B scavenger receptor type I in phagocytosis of apoptotic rat spermatogenic cells by Sertoli cells. J Biol Chem 274(9):5901–5908

    Article  CAS  PubMed  Google Scholar 

  32. Kawasaki Y, Nakagawa A, Nagaosa K et al (2002) Phosphatidylserine binding of class B scavenger receptor type I, a phagocytosis receptor of testicular Sertoli cells. J Biol Chem 277:27559–27566

    Article  CAS  PubMed  Google Scholar 

  33. Lu Q, Gore M, Zhang Q et al (1999) Tyro-3 family receptors are essential regulators of mammalian spermatogenesis. Nature 398:723–728

    Article  CAS  PubMed  Google Scholar 

  34. Xiong W, Chen Y, Wang H et al (2008) Gas6 and the Tyro 3 receptor tyrosine kinase subfamily regulate the phagocytic function of Sertoli cells. Reproduction 135:77–87

    Article  CAS  PubMed  Google Scholar 

  35. Chen Y, Wang H, Qi N et al (2009) Functions of TAM RTKs in regulating spermatogenesis and male fertility in mice. Reproduction 138:655–666

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We specially thank Drs Michel Blanc (Ret.), Jean-Louis Dacheux (Ret.), and Rustem Uzbekov (Plateforme IBiSA de Microscopie Electronique, Université François Rabelais and CHU de Tours, France) for the help in setting on simplified methods of phagocytosis rate analysis in routine experiments. We thank Image-UP platform from Poitiers University and Imaging & Microscopy Technology Platform from IGBMC, Strasbourg.

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Authors and Affiliations

  1. Laboratoire Signalisation et Transports Ioniques Membranaires, Université de Poitiers/CNRS, Poitiers, France

    Marina G. Yefimova, Annie-Claire Meunier, Anne Cantereau & Nicolas Bourmeyster

  2. Sechenov Institute of Evolutionary Physiology and Biochemistry, Russian Academy of Sciences, St. Petersbourg, Russia

    Marina G. Yefimova

  3. Institute of Genetics and Molecular and Cellular Biology, Illkirch, France

    Nadia Messaddeq

  4. Centre National de la Recherche Scientifique, UMR7104, Illkirch, France

    Nadia Messaddeq

  5. Institut National de la Santé et de la Recherche Médicale, U964, Illkirch, France

    Nadia Messaddeq

  6. Université de Strasbourg, Strasbourg, France

    Nadia Messaddeq

  7. Plateforme IMAGE-UP, Poitiers, France

    Anne Cantereau

  8. Université de Rennes/INSERM, IRSET, Rennes, France

    Bernard Jegou

  9. CHU de Poitiers, Poitiers, France

    Nicolas Bourmeyster

Authors
  1. Marina G. Yefimova
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  2. Nadia Messaddeq
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  3. Annie-Claire Meunier
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  4. Anne Cantereau
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  5. Bernard Jegou
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  6. Nicolas Bourmeyster
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Corresponding author

Correspondence to Nicolas Bourmeyster .

Editor information

Editors and Affiliations

  1. Department of Microscopy, Laboratory of Cell Biology and Unit for Multidisciplinary Research in Biomedicine (UMIB), Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto, Porto, Portugal

    Marco G. Alves

  2. Department of Microscopy, Laboratory of Cell Biology and Unit for Multidisciplinary Research in Biomedicine (UMIB), Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto, Porto, Portugal

    Pedro F. Oliveira

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Yefimova, M.G., Messaddeq, N., Meunier, AC., Cantereau, A., Jegou, B., Bourmeyster, N. (2018). Phagocytosis by Sertoli Cells: Analysis of Main Phagocytosis Steps by Confocal and Electron Microscopy. In: Alves, M., Oliveira, P. (eds) Sertoli Cells. Methods in Molecular Biology, vol 1748. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7698-0_8

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  • DOI: https://doi.org/10.1007/978-1-4939-7698-0_8

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7697-3

  • Online ISBN: 978-1-4939-7698-0

  • eBook Packages: Springer Protocols

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