Foxn1 and Prkdc genes are important for testis function: evidence from nude and scid adult mice

  • Carolina FA Oliveira
  • Nathália LM Lara
  • Samyra MSN Lacerda
  • Rodrigo R Resende
  • Luiz R França
  • Gleide F AvelarEmail author
Regular Article


Mutations in Foxn1 and Prkdc genes lead to nude and severe combined immunodeficiency (scid) phenotypes, respectively. Besides being immunodeficient, previous reports have shown that nude mice have lower gonadotropins and testosterone levels, while scid mice present increased pachytene spermatocyte (PS) apoptosis. Therefore, these specific features make them important experimental models for understanding Foxn1 and Prkdc roles in reproduction. Hence, we conducted an investigation of the testicular function in nude and scid BALB/c adult male mice and significant differences were observed, especially in Leydig cell (LC) parameters. Although the differences were more pronounced in nude mice, both immunodeficient strains presented a larger number of LC, whereas its cellular volume was smaller in comparison to the wild type. Besides these alterations in LC, we also observed differences in androgen receptor and steroidogenic enzyme expression in nude and scid mice, suggesting the importance of Foxn1 and Prkdc genes in androgen synthesis. Specifically in scid mice, we found a smaller meiotic index, which represents the number of round spermatids per PS, indicating a greater cell loss during meiosis, as previously described in the literature. In addition and for the first time, Foxn1 was identified in the testis, being expressed in LC, whereas DNA-PKc (the protein produced by Prkdc) was observed in LC and Sertoli cells. Taken together, our results show that the changes in LC composition added to the higher expression of steroidogenesis-related genes in nude mice and imply that Foxn1 transcription factor may be associated to androgen production regulation, while Prkdc expression is also important for the meiotic process.


Nude mice Scid mice Leydig cell Foxn1 Prkdc 



Technical assistance from Mara Lívia dos Santos is highly appreciated.

Funding information

This work was supported by the Brazilian National Council for Scientific and Technological Development (CNPq), the Foundation for Research Support of Minas Gerais (FAPEMIG) and the Coordination for the Improvement of Higher Education Personnel (CAPES).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in studies involving animals were in accordance with the ethical standards of the institution at which the studies were conducted (Ethics Committee in Animal Experimentation of the Federal University of Minas Gerais—CETEA/UFMG—Protocol no. 123/2013).


  1. Abercrombie M (1946) Estimation of nuclear populations from microtome sections. Anat Rec 94:238–248CrossRefGoogle Scholar
  2. Amann RP, Almquist JO (1962) Reproductive capacity of dairy bulls. Direct and indirect measurement of testicular sperm production. J Dairy Sci 45:774–781CrossRefGoogle Scholar
  3. Attal J, Courot M (1963) Development testiculaire et etablissement de la spermatogenese chez le taureau. Ann Biol Anim Biochem Biophys 8:219–241CrossRefGoogle Scholar
  4. Auharek SA, Avelar GF, Lara NL, Sharpe RM, França LR (2011) Sertoli cell numbers and spermatogenic efficiency are increased in inducible nitric oxide synthase mutant mice. Int J Androl. CrossRefGoogle Scholar
  5. Blunt T, Gell D, Fox M, Taccioli GE, Lehmann AR, Jackson SP, Jeggo PA (1996) Identification of a nonsense mutation in the carboxyl-terminal region of DNA-dependent protein kinase catalytic subunit in the scid mouse. Proc Natl Acad Sci 93:10285–10290CrossRefGoogle Scholar
  6. Brissette JL, Li J, Kamimura J, Lee D, Dotto GP (1996) The product of the mouse nude locus, Whn, regulates the balance between epithelial cell growth and differentiation. Genes Dev 10:2212–2221CrossRefGoogle Scholar
  7. Buzzard JJ, Farnworth PG, De Kretser DM, O’Connor AE, Wreford NG, Morrison JR (2003) Proliferative phase sertoli cells display a developmentally regulated response to activin in vitro. Endocrinology. CrossRefGoogle Scholar
  8. Byrd LG (1993) Regional localization of the nu mutation on mouse chromosome 11. Immunogenetics 37:157–159CrossRefGoogle Scholar
  9. De Rooij DG, De Boer P (2003) Specific arrest of spermatogenesis in genetically modified and mutant mice. Cytogenet Genome Res.
  10. Desjardins C, Ewing LL (1993) Cell and molecular biology of the testis, 1st edn. Oxford University Press, New YorkGoogle Scholar
  11. Dornas RA, Oliveira AG, Kalapothakis E, Hess RA, Mahecha GA, Oliveira CA (2007) Distribution of vitamin D3 receptor in the epididymal region of roosters (Gallus domesticus) is cell and segment specific. Gen Comp Endocrinol. CrossRefGoogle Scholar
  12. Dornas RA, Oliveira AG, Dias MO, Mahecha GA, Oliveira CA (2008) Comparative expression of androgen receptor in the testis and epididymal region of roosters (Gallus domesticus) and drakes (Anas platyrhynchos). Gen Comp Endocrinol. CrossRefGoogle Scholar
  13. Dorst VJ, Sajonski H (1974) Morphometrische Untersuchunhen am Tubulussys-tem des Schweinehodens wahrend der postnatalen Entwicklug. Monaths Vet Med 29:650–652Google Scholar
  14. Franca LR. (1992) Daily sperm production in Piau boars estimated from Sertoli cell population and Sertoli cell index. In: Proceedings of the 12th International Congress on animal reproduction and artificial insemination, vol. 4. The hague: The ICAR (ed. SJ Dieleman), pp. 1716–1718. Elsevier Science, The HagueGoogle Scholar
  15. Genissel C, Levallet J, Carreau S (2001) Regulation of cytochrome P450 aromatase gene expression in adult rat Leydig cells: comparison with estradiol production. J Endocrinol 168:95–105CrossRefGoogle Scholar
  16. Gholami S, Ansari-lari M, Khalili L (2015) Histologic and histomorphometric changes of testis following oral exposure to methyl tertiary-butyl ether in adult rat. Iran J Vet Res 16(3):288–292PubMedPubMedCentralGoogle Scholar
  17. Goertz MJ, Wu Z, Gallardo TD, Hamra FK, Castrillon DH (2011) Foxo1 is required in mouse spermatogonial stem cells for their maintenance and the initiation of spermatogenesis. J Clin Invest. CrossRefGoogle Scholar
  18. Hamer G, Roepers-Gajadien HL, Van Duyn-Goedhart A, Gademan IS, Kal HB, Van Buul PP, Ashley T, De Rooij DG (2003) Function of DNA-protein kinase catalytic subunit during the early meiotic prophase without Ku70 and Ku86. Biol Reprod 68:717–721CrossRefGoogle Scholar
  19. Hochereau-de Reviers MT, Lincoln GA (1978) Seasonal variation in the histology of the testis of the red deer, Cervus elephus. J Reprod Fertil 54:209–213CrossRefGoogle Scholar
  20. Jasurda JS, Jung DO, Froeter ED, Schwartz DB, Hopkins TD, Farris CL, McGee S, Narayan P, Ellsworth BS (2014) The forkhead transcription factor, FOXP3: a critical role in male fertility in mice. Biol Reprod.
  21. Justice NJ, Blount AL, Pelosi E, Schlessinger D, Vale W, Bilezikjian LM (2011) Impaired FSHβ expression in the pituitaries of Foxl2 mutant animals. Mol Endocrinol. CrossRefGoogle Scholar
  22. Keeney DS, Mendis-Handagama SMLC, Zirkin BR, Ewing LL (1988) Effect of long term deprivation of luteinizing hormone on Leydig cell volume, Leydig cell number and steroidogenic capacity of the rat testis. Endocrinology. CrossRefGoogle Scholar
  23. Lara NLM, Costa GMJ, Avelar GF, Lacerda SMSN, Hess RA, França LR (2018) Testis physiology—overview and histology. In: Skinner MK (ed) Encyclopedia of reproduction. Academic, Elsevier, pp 105–116CrossRefGoogle Scholar
  24. Lees-Miller SP, Meek K (2003) Repair of DNA double strand breaks by non-homologous end joining. Biochimie 85:1161–1173CrossRefGoogle Scholar
  25. Lucas TF, Nascimento AR, Pisolato R, Pimenta MT, Lazari MF, Porto CS (2014) Receptors and signaling pathways involved in proliferation and differentiation of Sertoli cells. Spermatogenesis. CrossRefGoogle Scholar
  26. Meachem SJ, Ruwanpura SM, Ziolkowski J, Ague JM, Skinner MK, Loveland KL (2005) Developmentally distinct in vivo effects of FSH on proliferation and apoptosis during testis maturation. J Endocrinol. CrossRefGoogle Scholar
  27. Mecklenburg L, Tychsen B, Paus R (2005) Learning from nudity: lessons from the nude phenotype. Exp Dermatol. CrossRefGoogle Scholar
  28. Meroni SB, Galardo MN, Rindone G, Gorga A, Riera MF, Cigorraga SB (2019) Molecular mechanisms and signaling pathways involved in Sertoli cell proliferation. Front Endocrinol.
  29. Morais RDVS, Crespo D, Nóbrega RH, Lemos MS, van de Kant HJG, de França LR, Male R, Bogerd J, Schulz RW (2017) Antagonistic regulation of spermatogonial differentiation in zebrafish (Danio rerio) by Igf3 and Amh. Mol Cell Endocrinol. CrossRefGoogle Scholar
  30. Nehls M, Pfeifer D, Schorpp M, Hedrich H, Boehm T (1994) New member of the winged-helix protein family disrupted in mouse and rat nude mutations. Nature. CrossRefGoogle Scholar
  31. Oliveira RL, Campolina-Silva GH, Nogueira JC, Mahecha GA, Oliveira CA (2013) Differential expression and seasonal variation on aquaporins 1 and 9 in the male genital system of big fruit-eating bat Artibeus lituratus. CrossRefGoogle Scholar
  32. Pannetier M, Fabre S, Batista F, Kocer A, Renault L, Jolivet G, Mandon-Pépin B, Cotinot C, Veitia R, Pailhoux E (2006) FOXL2 activates P450 aromatase gene transcription: towards a better characterization of the early steps of mammalian ovarian development. J Mol Endocrinol. CrossRefGoogle Scholar
  33. Pisarska MD, Bae J, Klein C, Hsueh AJ (2004) Forkhead l2 is expressed in the ovary and represses the promoter activity of the steroidogenic acute regulatory gene. Endocrinology. CrossRefGoogle Scholar
  34. Rebar RW, Morandini IC, Petze JE, Erickson GF (1982) Hormonal basis of reproductive defects in athymic mice: reduced gonadotropins and testosterone in males. Biol Reprod 27:1267–1276CrossRefGoogle Scholar
  35. Sampaio IMB (2002) Statistic applied to animal experimentation. 2nd. FEPMVZ, Belo HorizonteGoogle Scholar
  36. Scott HM, Hutchison GR, Jobling MS, McKinnell C, Drake AJ, Sharpe RM (2008) Relationship between androgen action in the “male programming window,” fetal sertoli cell number, and adult testis size in the rat. Endocrinology. CrossRefGoogle Scholar
  37. Skarra DV, Arriola DJ, Benson CA, Thackray VG (2013) Forkhead box O1 is a repressor of basal and GnRH-induced Fshb transcription in gonadotropes. Mol Endocrinol. CrossRefGoogle Scholar
  38. Sprando RL, Santulli R, Awoniyi CA, Ewing LL, Zirkin BR (1990) Does ethane 1,2-dimethanesulphonate (EDS) have a direct cytotoxic effect on the seminiferous epithelium of the rat testis? J Androl 11:344–352PubMedGoogle Scholar
  39. Tan J, Joseph DR, Quarmby VE, Lubahn DB, Sar M, French FS, Wilson EM (1988) The rat androgen receptor: primary structure, autoregulation of its messenger ribonucleic acid, and immunocytochemical localization of the receptor protein. Mol Endocrinol. CrossRefGoogle Scholar
  40. Tan KA, De Gendt K, Atanassova N, Walker M, Sharpe RM, Saunders PT, Denolet E, Verhoeven G (2005) The role of androgens in sertoli cell proliferation and functional maturation: studies in mice with total or Sertoli cell-selective ablation of the androgen receptor. Endocrinology. CrossRefGoogle Scholar
  41. Thackray VG (2014) Fox tales: regulation of gonadotropin gene expression by forkhead transcription factors. Mol Cell Endocrinol. CrossRefGoogle Scholar
  42. Uhlenhaut NH, Treier M (2011) Forkhead transcription factors in ovarian function. Reproduction. CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Laboratory of Cellular Biology, Department of MorphologyFederal University of Minas GeraisBelo HorizonteBrazil
  2. 2.Department of Biochemistry and ImmunologyFederal University of Minas GeraisBelo HorizonteBrazil

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