Assessment of Sperm Morphology, Chromatin Integrity, and Catsper Genes Expression in Hypothyroid Mice

Abstract

There is an evident relationship between the fertilizing capacity of sperm and the normal morphology, quality chromatin, and motility of sperm. It is well known that thyroid hormones are the important regulators of testicular function. A correlation was found between the hypothyroidism and sperm damages. The present study was conducted to investigate the effects of hypothyroidism on sperm morphology, chromatin quality, and motility. For this purpose, 20 male mice were divided into the control and the hypothyroid groups that received 0.05% 6-n-propyl-2-thiouracil (PTU) for 35 days. Sperm morphology with Papanicolaou staining and sperm chromatin quality with both Aniline Blue (AB) and Toluidine blue (TB) staining were assessed. Besides, immunohistochemistry and real-time PCR were performed to evaluate the changes of cation sperm channel (CatSper) genes. A significant increase in the sperm chromatin condensation was found in the hypothyroid mice compared to the control mice (p < 0.05). Furthermore, a significant decrease was observed in the morphology of normal sperm in hypothyroid mice compared to the controls (p < 0.05). The results showed that Hypothyroidism could downregulate the expression of CatSper genes. Immunohistochemical data confirmed the real time-PCR results. Furthermore, the results showed that hypothyroidism could adversely affect sperm morphology, sperm chromatin condensation, and CatSper gene expression in mice and these abnormalities may be related to the excessive production of reactive oxygen species (ROS) in a hypothyroid state.

References

  1. 1.

    Alasmari, W., Barratt, C. L., Publicover, S. J., Whalley, K. M., Foster, E., Kay, V., Martins da Silva, S., Oxenham, S. K. (2013) The clinical significance of calcium-signalling pathways mediating human sperm hyperactivation. Hum. Reprod. 28, 866–876.

    CAS  PubMed  PubMed Central  Google Scholar 

  2. 2.

    Amini, M., Shirinbayan, P., Behnam, B., Roghani, M., Farhoudian, A., Joghataei, M. T., Koruji, M. (2014) Correlation between expression of CatSper family and sperm profiles in the adult mouse testis following Iranian Kerack abuse. Andrology 2, 386–393.

    CAS  PubMed  Google Scholar 

  3. 3.

    Anbalagan, J., Sashi, A. M., Vengatesh, G., Stanley, J. A., Neelamohan, R., Aruldhas, M. M. (2010) Mechanism underlying transient gestational-onset hypothyroidism-induced impairment of posttesticular sperm maturation in adult rats. Fertil. Steril. 93, 2491–2497.

    CAS  PubMed  Google Scholar 

  4. 4.

    Bakhtiary, Z., Shahrooz, R., Ahmadi, A., Zarei, L. (2015) Protective effects of ethyl pyruvate on sperm quality in cyclophosphamide treated mice. Iran J. Reprod. Med. 13, 291–296.

    PubMed Central  Google Scholar 

  5. 5.

    Canale, D., Agostini, M., Giorgilli, G., Caglieresi, C., Scartabelli, G., Nardini, V., Jannini, E. A., Martino, E., Pinchera, A., Macchia, E. (2001) Thyroid hormone receptors in neonatal, prepubertal, and adult rat testis. J. Androl. 22, 284–288.

    CAS  PubMed  Google Scholar 

  6. 6.

    Chattopadhyay, S., Choudhury, S., Roy, A., Chainy, G. B., Samanta, L. (2010) T3 fails to restore mitochondrial thiol redox status altered by experimental hypothyroidism in rat testis. Gen. Comp. Endocrinol. 169, 39–47.

    CAS  PubMed  Google Scholar 

  7. 7.

    Choudhury, S., Chainy, G. B., Mishro, M. M. (2003) Experimentally induced hypo- and hyper-thyroidism influence on the antioxidant defence system in adult rat testis. Andrologia 35, 131–140.

    CAS  Google Scholar 

  8. 8.

    Chung, J. J., Navarro, B., Krapivinsky, G., Krapivinsky, L., Clapham, D. E. (2011) A novel gene required for male fertility and functional CATSPER channel formation in spermatozoa. Nat. Commun. 2, 153.

    PubMed  PubMed Central  Google Scholar 

  9. 9.

    Dittrich, R., Beckmann, M. W., Oppelt, P. G., Hoffmann, I., Lotz, L., Kuwert, T., Mueller, A. (2011) Thyroid hormone receptors and reproduction. J. Reprod. Immunol. 90, 58–66.

    CAS  PubMed  Google Scholar 

  10. 10.

    Gao, Y., Lee, W. M., Cheng, C. Y. (2014) Thyroid hormone function in the rat testis. Front. Endocrinol. (Lausanne) 5, 188.

    Google Scholar 

  11. 11.

    Heidari, R., Alizadeh, R., Abbasi, N., Pasbakhsh, P., Hedayatpour, A., Farajpour, M., Khaleghi, M. R., Abbasi, M., Dehpour, A. R. (2015) Do pilea microphylla improve sperm DNA fragmentation and sperm parameters in varicocelized rats? Acta Med. Iran 53, 547–554.

    PubMed  Google Scholar 

  12. 12.

    Jin, J., Jin, N., Zheng, H., Ro, S., Tafolla, D., Sanders, K. M., Yan, W. (2007) Catsper3 and Catsper4 are essential for sperm hyperactivated motility and male fertility in the mouse. Biol. Reprod. 77, 37–44.

    CAS  PubMed  Google Scholar 

  13. 13.

    Kirichok, Y., Lishko, P. V. (2011) Rediscovering sperm ion channels with the patch-clamp technique. Mol. Hum. Reprod. 17, 478–499.

    CAS  PubMed  PubMed Central  Google Scholar 

  14. 14.

    Krajewska-Kulak, E., Sengupta, P. (2013) Thyroid function in male infertility. Front. Endocrinol. (Lausanne) 4, 174.

    Google Scholar 

  15. 15.

    Krassas, G. E., Papadopoulou, F., Tziomalos, K., Zeginiadou, T., Pontikides, N. (2008) Hypothyroidism has an adverse effect on human spermatogenesis: a prospective, controlled study. Thyroid. 18, 1255–1259.

    CAS  PubMed  Google Scholar 

  16. 16.

    Krassas, G. E., Poppe, K., Glinoer, D. (2010) Thyroid function and human reproductive health. Endocr. Rev. 31, 702–755.

    CAS  PubMed  Google Scholar 

  17. 17.

    Kumar, A., Shekhar, S., Dhole, B. (2014) Thyroid and male reproduction. Indian J. Endocrinol. Metab. 18, 23–31.

    CAS  PubMed  PubMed Central  Google Scholar 

  18. 18.

    Mangoli, E., Talebi, A. R., Anvari, M., Pourentezari, M. (2013) Effects of experimentally-induced diabetes on sperm parameters and chromatin quality in mice. Iran J. Reprod. Med. 11, 53–60.

    CAS  PubMed  PubMed Central  Google Scholar 

  19. 19.

    Mendeluk, G. R., Rosales, M. (2016) Thyroxin is useful to improve sperm motility. Int. J. Fertil Steril. 10, 208–214.

    CAS  PubMed  PubMed Central  Google Scholar 

  20. 20.

    Mohammadi, S., Jalali, M., Nikravesh, M. R., Fazel, A., Ebrahimzadeh, A., Gholamin, M., Sankian, M. (2013) Effects of vitamin-E treatment on CatSper genes expression and sperm quality in the testis of the aging mouse. Iran J. Reprod. Med. 11, 989–998.

    CAS  PubMed  PubMed Central  Google Scholar 

  21. 21.

    Mohammadi, S., Movahedin, M., Mowla, S. J. (2009) Up-regulation of CatSper genes family by selenium. Reprod. Biol. Endocrinol. 7, 126.

    PubMed  PubMed Central  Google Scholar 

  22. 22.

    Nazar, M., Talebi, A. R., Hosseini Sharifabad, M., Abbasi, A., Khoradmehr, A., Danafar, A. H. (2016) Acute and chronic effects of gold nanoparticles on sperm parameters and chromatin structure in mice. Int. J. Reprod. Biomed. (Yazd) 14, 637–642.

    Google Scholar 

  23. 23.

    Nikoobakht, M. R., Aloosh, M., Nikoobakht, N., Mehrsay, A. R., Biniaz, F., Karjalian, M. A. (2012) The role of hypothyroidism in male infertility and erectile dysfunction. Urol. J. 9, 405–409.

    PubMed  Google Scholar 

  24. 24.

    Nikpoor, P., Mowla, S. J., Movahedin, M., Ziaee, S. A., Tiraihi, T. (2004) CatSper gene expression in postnatal development of mouse testis and in subfertile men with deficient sperm motility. Hum. Reprod. 19, 124–128.

    PubMed  Google Scholar 

  25. 25.

    Park, E. H., Kim, D. R., Kim, H. Y., Park, S. K., Chang, M. S. (2014) Panax ginseng induces the expression of CatSper genes and sperm hyperactivation. Asian J. Androl. 16, 845–851.

    PubMed  PubMed Central  Google Scholar 

  26. 26.

    Pourentezari, M., Talebi, A., Abbasi, A., Khalili, M. A., Mangoli, E., Anvari, M. (2014) Effects of acrylamide on sperm parameters, chromatin quality, and the level of blood testosterone in mice. Iran J. Reprod. Med. 12, 335–342.

    CAS  PubMed  PubMed Central  Google Scholar 

  27. 27.

    Qi, H., Moran, M. M., Navarro, B., Chong, J. A., Krapivinsky, G., Krapivinsky, L., Kirichok, Y., Ramsey, I. S., Quill, T. A., Clapham, D. E. (2007) All four CatSper ion channel proteins are required for male fertility and sperm cell hyperactivated motility. Proc. Natl Acad. Sci. USA 104, 1219–1223.

    CAS  PubMed  Google Scholar 

  28. 28.

    Quill, T. A., Ren, D., Clapham, D. E., Garbers, D. L. (2001) A voltage-gated ion channel expressed specifically in spermatozoa. Proc. Natl Acad. Sci. USA 98, 12527–12531.

    CAS  PubMed  Google Scholar 

  29. 29.

    Ren, D., Navarro, B., Perez, G., Jackson, A. C., Hsu, S., Shi, Q., Tilly, J. L., Clapham, D. E. (2001) A sperm ion channel required for sperm motility and male fertility. Nature 413, 603–609.

    CAS  PubMed  Google Scholar 

  30. 30.

    Roodbari, F., Abedi, N., Talebi, A. R. (2015) Early and late effects of ibuprofen on mouse sperm parameters, chromatin condensation, and DNA integrity in mice. Iran J. Reprod. Med. 13, 703–710.

    PubMed Central  Google Scholar 

  31. 31.

    Sabour, M., Khoradmehr, A., Kalantar, S. M., Danafar, A. H., Omidi, M., Halvaei, I., Nabi, A., Ghasemi-Esmailabad, S., Talebi, A. R. (2017) Administration of high dose of methamphetamine has detrimental effects on sperm parameters and DNA integrity in mice. Int. J. Reprod. Biomed. (Yazd) 15, 161–168.

    CAS  Google Scholar 

  32. 32.

    Sahoo, D. K., Roy, A. (2012) Compromised rat testicular antioxidant defence system by hypothyroidism before puberty. Int. J. Endocrinol. 2012, 637825.

    PubMed  PubMed Central  Google Scholar 

  33. 33.

    Sahoo, D. K., Roy, A., Bhanja, S., Chainy, G. B. (2008) Hypothyroidism impairs antioxidant defence system and testicular physiology during development and maturation. Gen. Comp. Endocrinol. 156, 63–70.

    CAS  PubMed  Google Scholar 

  34. 34.

    Sasikumar, S., Dakshayani, D. (2013) Assessment of sperm DNA integrity by Toluidine blue staining technique in infertile patients and its relation to cryopreservation. Int. J. Curr. Microbiol. App. Sci. 2, 280–292.

    Google Scholar 

  35. 35.

    Shukla, K. K., Mahdi, A. A., Rajender, S. (2012) Ion channels in sperm physiology and male fertility and infertility. J. Androl. 33, 777–788.

    CAS  PubMed  Google Scholar 

  36. 36.

    Singh, A. P., Rajender, S. (2015) CatSper channel, sperm function and male fertility. Reprod. Biomed. Online 30, 28–38.

    PubMed  Google Scholar 

  37. 37.

    Singh, R., Hamada, A., Agarwal, A. (2011) Thyroid hormones in male reproduction and fertility. Open Reprod. Sci. J. 3, 98–104.

    CAS  Google Scholar 

  38. 38.

    Talebi, A. R., Mangoli, E., Nahangi, H., Anvari, M., Pourentezari, M., Halvaei, I. (2014) Vitamin C attenuates detrimental effects of diabetes mellitus on sperm parameters, chromatin quality and rate of apoptosis in mice. Eur. J. Obstet. Gynecol. Reprod. Biol. 181, 32–36.

    CAS  PubMed  Google Scholar 

  39. 39.

    Talebi, A. R., Vahidi, S., Aflatoonian, A., Ghasemi, N., Ghasemzadeh, J., Firoozabadi, R. D., Moein, M. R. (2012) Cytochemical evaluation of sperm chromatin and DNA integrity in couples with unexplained recurrent spontaneous abortions. Andrologia 44 Suppl. 1, 462–470.

    Google Scholar 

  40. 40.

    Ulisse, S., Rucci, N., Piersanti, D., Carosa, E., Graziano, F. M., Pavan, A., Ceddia, P., Arizzi, M., Muzi, P., Cironi, L., Gnessi, L., D’Armiento, M., Jannini, E. A. (1998) Regulation by thyroid hormone of the expression of basement membrane components in rat prepubertal Sertoli cells. Endocrinology 139, 741–747.

    CAS  PubMed  Google Scholar 

  41. 41.

    Wagner, M. S., Wajner, S. M., Maia, A. L. (2008) The role of thyroid hormone in testicular development and function. J. Endocrinol. 199, 351–365.

    CAS  PubMed  PubMed Central  Google Scholar 

  42. 42.

    Wajner, S. M., Wagner, M. S., Maia, A. L. (2009) Clinical implications of altered thyroid status in male testicular function. Arq. Bras. Endocrinol. Metabol. 53, 976–982.

    PubMed  Google Scholar 

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Correspondence to Mehdi Jalali.

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Alipour, F., Jalali, M., Nikravesh, M.R. et al. Assessment of Sperm Morphology, Chromatin Integrity, and Catsper Genes Expression in Hypothyroid Mice. BIOLOGIA FUTURA 69, 244–258 (2018). https://doi.org/10.1556/018.68.2018.3.2

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Key words

  • hypothyroidism
  • sperm
  • chromatin
  • CatSper
  • testis