Study on association of four polymorphisms in TNP2 gene with spermatogenesis and causes of male infertility

Subject Areas : Developmental biology of plants and animals , development and differentiation in microorganisms

elham siasi ¹ , Ahmad Aleyasin ² , Javad Mowla ³

1 - Department of Microbiology, Faculty of science, North Tehran Branch, Islamic Azad University, Tehran, Iran.
2 - Medical genetic department, National Institute for Genetic Engineering and Biotechnology, Tehran, Iran.
3 - Molecular Genetic Department, Tarbiat Modares University, Tehran, Iran.

Received: 2016-01-04 Accepted : 2016-02-24 Published : 2016-05-21

Keywords: PCR-RFLP, PCR-SSCP, SNP, Male infertility, TNP2 gene,

Abstract :

Protamines are DNA-binding proteins in the sperm nucleus, which cause differentiation of human spermatogenesis. Protamines required for packaging nucleus sperm by replacement of histones with protamines. Sperm protamine deficiency has been associated with spermatogenesis failed and caused male infertility.  In this study association of four SNPs in TNP2 gene were studied in Iranian idiopathic infertile men with azoospermia or oligospermia. Analysis of four SNPs include T1019G, G1272C،  G deletion at 1036 and 1046 in TNP2 gene  was performed by DNA extraction from blood samples of 96 idiopathic infertile men with azoospermia or oligospermia and 100 normal control men. Then using restriction fragment length polymorphism (PCR-RFLP) for detection of T1019G and G1272C, SNPs and for identification of G deletion at 1036 and 1046 used Single Strand Conformation Polymorphism (PCR- SSCP). Results were confirmed by DNA sequencing analysis. For (G1272C) in TNP2 gene, frequency of CC genotype was differented in fertile and infertile groups but statistical analysis showed no significant association related to this SNP in case and control groups. As also no polymorphisms were found for T1019G and G deletion at 1036 and 1046 nt. These results are consistent with previous studies and indicating that four tested SNPs was not associated with oligospermia and azospermia and idiopatic male infertility in Iranian population.

References:

منابع

[1]     Aston K.I., Krausz C., Laface I., Ruiz-Castane E.D.T. 2010. Evaluation of 172 candidate polymorphisms for association with oligozoospermia or azoospermia in a large cohort of men of European descent. Human Reproduction 2010, 5: 1-15.

[2]      Dada R., Thilagavathi J., Venkatesh S., Esteves S.C., Agarwa A. 2011. Genetic Testing in Male Infertility. The Open Reproductive Science Journal, 3: 42-56.

[3]      El-Toukhy T.P.B. 2002. Male infertility and ICSI. Current Obstetrics and Gynaecology, 12: 276-285.

[4]     Ferlin A., Arredi B.C.F. 2006. Genetic causes of male infertility. Reprod Toxicol, 22: 133-141.

[5]      Gazouez C., Oriola J., De Mateo S., Vidal-Taboada J.M., Ballesca J.I. 2008. A comman protamine 1 promoter polymorphism (C190A) correlates with abnormal sperm morphology and increased protamine P1/P2 ratio in infertile patients. Journal of Andrology, 29: 540-548.

[6]      Heidari M.M., Khatami M., Talebi A.R., Moezzi F. 2014. Mutation analysis of  TNP1 gene in infertile men with varicocele. Iran J Reprod, 12(4): 257-262.

[7]     Hogarth C., Itman C., Jans D.A. 2005. Regulated nucleocytoplasmic transport in spermatogenesis: a driver of cellular differentiation? Bioessays, 27: 1011-1025.

[8]      Iguchi N., Yang S., Lamb D.J. 2006. An SNP in protamine 1: a possible genetic cause of male infertility? J Med Genet, 43: 382-384.

[9]      Imken L., Rouba H., EI Houate B., Louanjli N., Barakat A., Chafik A. 2009.  Mutations in the protamine locus: association with spermatogenic failure? Molecular Human Reproduction, 7: 1-22.

[10]  Jeffreys A.J. 2013. The man behind the DNA fingerprints: an interview with Professor Sir Alec Jeffreys. Investigative Genetics, 4(21): 1-7.

[11]  Katherine L., Varghes A.C., Agarwa A. 2010. The genetic causes of male factor infertility. Fertility and Sterility, 93(1): 1-12.

[12] Kempisty B., Depa-Martynow M., Lianeri M., Jedrzegczak P., Darul-Wasowicz A. 2007. Evaluation of protamines 1 and 2 transcript contents in spermatozoa from asthenozoospermic men. Folia Histochemica ET Cytobiologica, 45: 109-113.

[13]  Konstantinos K.V., Panagiotis P., Antonios V.T., Agelos P., Argiris N.V. 2008. PCR–SSCP: A Method for the Molecular Analysis of Genetic Diseases. Mol Biotechnol, 38: 155–163.

[14]  Lewin J.D., Abbott D.W.J. 2003. A haploid affair: core histone transitions during spermatogenesis. Biochem Cell Biol,, 81: 131-140.

[15]  Miyagawa Y., Nishimura H., Tsujimura A., Matsuoka Y., Matsumiya K., Okuyama A., Nishimune Y.H.T. 2005. Single-nucleotide polymorphisms and mutation analyses of the TNP1 and TNP2 genes of fertile and infertile human male populations. Journal of Andrology, 26(6): 779-786.

[16]  Oliva R. 2006. Protamines and male infertility. Human Reproduction Update, 12(4): 417-435.

[17]  Park Sh. 2016. Genetic Factors and Environmental Factors Affecting Male Infertility. International Research Journal of Advanced Engineering and Science, 1(3): 115-118.

[18]  Poongothai J., Gopenath T.S., Manonayaki S. 2009.Genetics of human male infertility. Singapore Med J, 50(4): 336-347.

 

 

[19]   Raheem A.A., Ralph D. 2011, Male infertility: causes and investigations. Trends in urology and mens health, 2(5): 8-11.

[20]  Ravel C., Chantot-bastaraud S., EI Houate B., Berthaut I., Verstraete L., De Larouziere V., Lourenco D., Dumaine A, Antoine J.M., Mandelbaum J. 2007. Mutations in the protamine 1 gene associated with male infertility. Molecular Human Reproduction, 5: 1-4.

[21]  Shamsi M.B., Kumar K., Dada R. 2011.  Genetic and epigenetic factors: Role in male infertility. Indian Journal of Urology, 27(1): 110-120.

[22]  Tanaka H., Miyagawa Y., Tsujimura A., Matsumiya K., Okuyama A.Y.N. 2003. Single-nucleotide polymorphisms in the protamine-1 and -2 genes of fertile and infertile human male populations. Molecular Human Reproduction, 9: 69-73.

[23]  Venkatesh S., Kumar R., Deka D., Deecaraman M., Dada R. 2011. Analysis of sperm nuclear protein gene polymorphisms and DNA integrity in infertile men. Systems biology in reproductive medicine, 57(3): 124-132.

[24]  Vogt P.H. 2004. Molecular Genetic of Human Male Infertility: From Genes to New Therapeutic Perspectives. Current Pharmaceutical Design, 10(1): 1-30.