Journal of Reproduction & Infertility

Journal of Reproduction & Infertility

Expression Analysis of the CRISP2, CATSPER1, PATE1 and SEMG1 in the Sperm of Men with Idiopathic Asthenozoospermia

Authors
Zohreh Heidary 1
Majid Zaki-Dizaji 1
Kioomars Salimi Nejad 1
Hamid Reza Khorram Khorshid 2
1 Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran
2 Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran; Genetics Research Center, University of Social Welfare and Rehabilitation Sciences, Tehran, Iran
Abstract
Background: The purpose of this study was to analyze the expression level of CRISP2, CATSPER1, PATE1 and SEMG1 genes in the sperm of men with asthenozoospermia (AZS). AZS is a cause of infertility in men in which the motility of the sperm is reduced. So far, a few genes have been associated with AZS; however, in most of the cases, its molecular etiology is unclear. Methods: A total of 35 subjects with idiopathic AZS and 35 fertile and healthy men as control were included. In study after total RNA extraction and cDNA synthesis, relative quantification was performed. B2M was used as the normalizer gene and fold change was calculated by 2 −ΔΔCt method. Mann-Whitney test was used to compare the expression levels between the case and control groups with significance level of p<0.05. Results: Our results showed that CRISP2 (p=0.03) and SEMG1 (p=0.03) were significantly down- and up-regulated in AZS men respectively compared to the controls. But CATSPER1 and PATE1 did not show significant changes. Conclusion: Down-regulation of CRISP2 and up-regulation of SEMG1 were associated with AZS, which could be suggested as the potential candidate genes for the development of a diagnostic marker or potentially for more studies for treatment of AZS.
Keywords
Asthenozoospermia
CATSPER1
CRISP2
Gene expression
Male infertility
PATE1
real
time PCR
SEMG1
sperm motility
  1. Coutton C, Fissore RA, Palermo GD, Stouffs K, Toure A. Male infertility: genetics, mechanism, and therapies. Biomed Res Int. 2016;2016:7372362.
  2. WHO Organization. WHO laboratory manual for the examination and processing of human semen. 5th ed. Geneva: World health organization. 2010. 286 p.
  3. Ferlin A, Raicu F, Gatta V, Zuccarello D, Palka G, Foresta C. Male infertility: role of genetic background. Reprod Biomed Online. 2007;14(6):734-45.
  4. Tüttelmann F, Röpke A. Genetics of Male Infertility. In: Simoni M, Huhtaniemi IT, editors. Endocrinology of the Testis and Male Reproduction. Cham: Springer International Publishing; 2017. p. 1029-49.
  5. Miyamoto T, Minase G, Shin T, Ueda H, Okada H, Sengoku K. Human male infertility and its genetic causes. Reprod Med Biol. 2017;16(2):81-8.
  6. Hargreave TB, Elliott D. Genetics and male infertility. In: Schill WB, Comhaire FH, Hargreave TB, editors. Andrology for the clinician. Berlin, Heidelberg: Springer; 2006. p. 462-80.
  7. Neto FT, Bach PV, Najari BB, Li PS, Goldstein M. Genetics of male infertility. Curr Urol Rep. 2016;17(10):70.
  8. Walsh TJ, Pera RR, Turek PJ. The genetics of male infertility. Semin Reprod Med. 2009;27(2):124-36.
  9. Asero P, Calogero AE, Condorelli RA, Mongioi L, Vicari E, Lanzafame F, et al. Relevance of genetic investigation in male infertility. J Endocrinol Invest. 2014;37(5):415-27.
  10. Curi SM, Ariagno JI, Chenlo PH, Mendeluk GR, Pugliese MN, Sardi Segovia LM, et al. Asthenozoospermia: analysis of a large population. Arch Androl. 2003;49(5):343-9.
  11. Heidary Z, Saliminejad K, Zaki-Dizaji M, Khorram Khorshid HR. Genetic aspects of idiopathic asthenozoospermia as a cause of male infertility. Hum Fertil (Camb). 2018:1-10.
  12. Luo S, Jia J, Hu H, Ma W, Jiao Y, Dong J. A Chinese herbal decoction can increase the intracellular Ca2+ concentration and CatSper1 expression in mouse sperm tails. Mol Med Rep. 2013;7(1):195-200.
  13. Zhou JH, Zhou QZ, Lyu XM, Zhu T, Chen ZJ, Chen MK, et al. The expression of cysteine-rich secretory protein 2 (CRISP2) and its specific regulator miR-27b in the spermatozoa of patients with asthenozoospermia. Biol Reprod. 2015;92(1):28.
  14. Liu FJ, Liu X, Han JL, Wang YW, Jin SH, Liu XX, et al. Aged men share the sperm protein PATE1 defect with young asthenozoospermia patients. Hum Reprod. 2015;30(4):861-9.
  15. Yu Q, Zhou Q, Wei Q, Li J, Feng C, Mao X. SEMG1 may be the candidate gene for idiopathic asthenozoospermia. Andrologia. 2014;46(2):158-66.
  16. Mizuki N, Sarapata DE, Garcia-Sanz JA, Kasahara M. The mouse male germ cell-specific gene Tpx-1: molecular structure, mode of expression in spermatogenesis, and sequence similarity to two non-mammalian genes. Mamm Genome. 1992;3(5):274-80.
  17. Kratzschmar J, Haendler B, Eberspaecher U, Roosterman D, Donner P, Schleuning WD. The human cysteine-rich secretory protein (CRISP) family. Primary structure and tissue distribution of CRISP-1, CRISP-2 and CRISP-3. Eur J Biochem. 1996;236(3):827-36.
  18. Haendler B, Habenicht UF, Schwidetzky U, Schuttke I, Schleuning WD. Differential androgen regulation of the murine genes for cysteine-rich secretory proteins (CRISP). Eur J Biochem. 1997;250(2):440-6.
  19. O'Bryan MK, Sebire K, Meinhardt A, Edgar K, Keah HH, Hearn MT, et al. Tpx-1 is a component of the outer dense fibers and acrosome of rat spermatozoa. Mol Reprod Dev. 2001;58(1):116-25.
  20. Busso D, Cohen DJ, Hayashi M, Kasahara M, Cuasnicu PS. Human testicular protein TPX1/CRISP-2: localization in spermatozoa, fate after capacitation and relevance for gamete interaction. Mol Hum Reprod. 2005;11(4):299-305.
  21. Gibbs GM, Bianco DM, Jamsai D, Herlihy A, Ristevski S, Aitken RJ, et al. Cysteine-rich secretory protein 2 binds to mitogen-activated protein kinase kinase kinase 11 in mouse sperm. Biol Reprod. 2007;77(1):108-14.
  22. Jamsai D, Bianco DM, Smith SJ, Merriner DJ, Ly-Huynh JD, Herlihy A, et al. Characterization of gametogenetin 1 (GGN1) and its potential role in male fertility through the interaction with the ion channel regulator, cysteine-rich secretory protein 2 (CRISP2) in the sperm tail. Reproduction. 2008;135(6):751-9.
  23. Jamsai D, Rijal S, Bianco DM, O'Connor AE, Merriner DJ, Smith SJ, et al. A novel protein, sperm head and tail associated protein (SHTAP), interacts with cysteine-rich secretory protein 2 (CRISP2) during spermatogenesis in the mouse. Biol Cell. 2009;102(2):93-106.
  24. Nimlamool W, Bean BS, Lowe-Krentz LJ. Human sperm CRISP2 is released from the acrosome during the acrosome reaction and re-associates at the equatorial segment. Mol Reprod Dev. 2013;80(6):488-502.
  25. Harper CV, Barratt CL, Publicover SJ. Stimulation of human spermatozoa with progesterone gradients to simulate approach to the oocyte. Induction of [Ca(2+)](i) oscillations and cyclical transitions in flagellar beating. J Biol Chem. 2004;279(44):46315-25.
  26. Da Ros VG, Munoz MW, Battistone MA, Brukman NG, Carvajal G, Curci L, et al. From the epididymis to the egg: participation of CRISP proteins in mammalian fertilization. Asian J Androl. 2015;17(5):711-5.
  27. Ren D, Navarro B, Perez G, Jackson AC, Hsu S, Shi Q, et al. A sperm ion channel required for sperm motility and male fertility. Nature. 2001;413(6856):603-9.
  28. Sanchez-Cardenas C, Montoya F, Navarrete FA, Hernandez-Cruz A, Corkidi G, Visconti PE, et al. Intracellular Ca2+ threshold reversibly switches flagellar beat off and on. Biol Reprod. 2018;99(5):1010-21.
  29. Ueda Y, Yamaguchi R, Ikawa M, Okabe M, Morii E, Maeda Y, et al. PGAP1 knock-out mice show otocephaly and male infertility. J Biol Chem. 2007;282(42):30373-80.
  30. Jensen-Seaman MI, Li WH. Evolution of the hominoid semenogelin genes, the major proteins of ejaculated semen. J Mol Evol. 2003;57(3):261-70.
  31. Lundwall A. A locus on chromosome 20 encompassing genes that are highly expressed in the epididymis. Asian J Androl. 2007;9(4):540-4.
  32. Zhou QZ, Guo XB, Zhang WS, Zhou JH, Yang C, Bian J, et al. Expressions of miR-525-3p and its target gene SEMG1 in the spermatozoa of patients with asthenozoospermia. Andrology. 2018.
  33. Moreno LI, Tate CM, Knott EL, McDaniel JE, Rogers SS, Koons BW, et al. Determination of an effective housekeeping gene for the quantification of mRNA for forensic applications. J Forensic Sci. 2012;57(4):1051-8.
  34. Cao X, Cui Y, Zhang X, Lou J, Zhou J, Bei H, et al. Proteomic profile of human spermatozoa in healthy and asthenozoospermic individuals. Reprod Biol Endocrinol. 2018;16(1):16.
  35. Zhou JH, Zhou QZ, Yang JK, Lyu XM, Bian J, Guo WB, et al. MicroRNA-27a-mediated repression of cysteine-rich secretory protein 2 translation in asthenoteratozoospermic patients. Asian J Androl. 2016;19(5):591-5.
  36. Li Y, Li M, Liu Y, Song G, Liu N. A microarray for microRNA profiling in spermatozoa from adult men living in an environmentally polluted site. Bull Environ Contam Toxicol. 2012;89(6):1111-4.
  37. Wang H, Zhou Z, Xu M, Li J, Xiao J, Xu ZY, et al. A spermatogenesis-related gene expression profile in human spermatozoa and its potential clinical applications. J Mol Med (Berl). 2004;82(5):317-24.
  38. Zhou J, Xue K, Chen M, Zhou Q, Yang J, Bian J, et al. [Expression of cysteine-rich secretory protein 2 in patients with asthenozoospermia and its clinical significance]. Nan Fang Yi Ke Da Xue Xue Bao. 2014;34(10):1528-33. Chinese.
  39. Jing XW, Xing RW, Zhou QZ, Yu QF, Guo WB, Chen SM, et al. [Expressions of cysteine-rich secretory protein 2 in asthenospermia]. Zhonghua Nan Ke Xue. 2011;17(3):203-7. Chinese.
  40. Du Y, Huang X, Li J, Hu Y, Zhou Z, Sha J. Human testis specific protein 1 expression in human spermatogenesis and involvement in the pathogenesis of male infertility. Fertil Steril. 2006;85(6):1852-4.
  41. Jamsai D, Reilly A, Smith SJ, Gibbs GM, Baker HW, McLachlan RI, et al. Polymorphisms in the human cysteine-rich secretory protein 2 (CRISP2) gene in Australian men. Hum Reprod. 2008;23(9):2151-9.
  42. Agarwal A, Sharma R, Durairajanayagam D, Ayaz A, Cui Z, Willard B, et al. Major protein alterations in spermatozoa from infertile men with unilateral varicocele. Reprod Biol Endocrinol. 2015;13:8.
  43. Martinez-Heredia J, de Mateo S, Vidal-Taboada JM, Ballesca JL, Oliva R. Identification of proteomic differences in asthenozoospermic sperm samples. Hum Reprod. 2008;23(4):783-91.
  44. Terai K, Yoshida K, Yoshiike M, Fujime M, Iwamoto T. Association of seminal plasma motility inhibitors/semenogelins with sperm in asthenozoospermia-infertile men. Urol Int. 2010;85(2):209-15.
  45. Legare C, Droit A, Fournier F, Bourassa S, Force A, Cloutier F, et al. Investigation of male infertility using quantitative comparative proteomics. J Proteome Res. 2014;13(12):5403-14.
  46. Robert M, Gagnon C. Semenogelin I: a coagulum forming, multifunctional seminal vesicle protein. Cell Mol Life Sci. 1999;55(6-7):944-60.
  47. Marques PI, Fonseca F, Carvalho AS, Puente DA, Damião I, Almeida V, et al. Sequence variation at KLK and WFDC clusters and its association to semen hyperviscosity and other male infertility phenotypes. Hum Reprod. 2016;31(12):2881-91.
  48. Tamburrino L, Marchiani S, Vicini E, Muciaccia B, Cambi M, Pellegrini S, et al. Quantification of CatSper1 expression in human spermatozoa and relation to functional parameters. Hum Reprod. 2015;30(7):1532-44.
  49. Shu F, Zhou X, Li F, Lu D, Lei B, Li Q, et al. Analysis of the correlation of CATSPER single nucleotide polymorphisms (SNPs) with idiopathic asthenospermia. J Assist Reprod Genet. 2015;32(11):1643-9.
  50. Wang YN, Wang B, Liang M, Han CY, Zhang B, Cai J, et al. Down-regulation of CatSper1 channel in epididymal spermatozoa contributes to the pathogenesis of asthenozoospermia, whereas up-regulation of the channel by Sheng-Jing-San treatment improves the sperm motility of asthenozoospermia in rats. Fertil Steril. 2013;99(2):579-87.
  51. Visser L, Westerveld GH, Xie F, van Daalen SK, van der Veen F, Lombardi MP, et al. A comprehensive gene mutation screen in men with asthenozoospermia. Fertil Steril. 2011;95(3):1020-4.e1-9.
  52. Avenarius MR, Hildebrand MS, Zhang Y, Meyer NC, Smith LL, Kahrizi K, et al. Human male infertility caused by mutations in the CATSPER1 channel protein. Am J Hum Genet. 2009;84(4):505-10.
  53. Zhang S, Wang QM, Ding XP, Wang T, Mu XM, Chen ZY. Association of polymorphisms in PATE1 gene with idiopathic asthenozoospermia in Sichuan, China. J Reprod Immunol. 2016;118:54-60.
Journal of Reproduction & Infertility
Volume 20, Issue 2
Spring 2019
Pages 70-75