Evaluation of outer dense fiber-1 and -2 protein expression in asthenozoospermic infertile men
Abstract
Background: Most of male infertility are caused by defect in sperm motility (asthenozoospermia). The molecular mechanism of low sperm motility in asthenozoospermic patients has not been fully understood. Sperm motility is strongly related to the axoneme structure which is composed of microtubules and supported by outer dense fiber (ODF) and fibrous sheath (FS) protein. The objective of this study was to characterize the ODF (ODF1 and ODF2) expression in asthenozoospermic infertile male and control normozoospermic fertile male.
Methods: Asthenozoospermic samples (n=18) were collected from infertile patients at Andrology Lab, Cipto Mangunkusumo Hospital Jakarta and control were taken from normozoospermic fertile donor (n=18). After motility analyses by computer-assisted sperm analysis (CASA), semen were divided into two parts, for Western blot and for immunocytochemistry analysis. Antibody against ODF1 and ODF2 protein were used in both analyses.
Results: Analysis of ODF1 protein expression showed bands with molecular weight of ~30 kDa and ODF2 ~85 kDa. The mean band intensity of ODF1 and ODF2 protein were lower in the asthenozoospermic group (AG) compared to normozoospermic group (NG). Moreover, both ODF proteins were less intense and less localized in the AG than NG. Sperm motility was lower in AG, compared to control NG, i.e. average path velocity (VAP) = 32.07 ± 7.03 vs 37.58 ± 8.73 µm/s, p = 0.455; straight line velocity (VSL) = 24.17 ± 6.90 vs 27.61 ± 4.50 µm/s, p = 0.317 and curvilinear velocity (VCL) = 45.68 ± 7.91 vs 55.55 ± 16.40 µm/s, p = 0.099.
Conclusion: There is down-regulation of ODF1 and ODF2 protein expression and less-compact localization in AG sperm compared to the NG. These changes might have caused disturbances in the sperm motility as observed in this study.
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References
Moeloek N, Ahda Y. Perbandingan kemanjuran dan keamanan beberapa preparat FSH untuk stimulasi ovarium perempuan yang mengikuti program reproduksi berbantuan. Maj Kedokteran Indonesia. 2001;51(12):450-4. Indonesian.
Hamada A, Esteves SC, Agarwal A. Unexplained male infertility: potential causes and management. Human Andrology. 2011;1:2-16. http://dx.doi.org/10.1097/01.XHA.0000397686.82729.09
Chuang WW, Lo KC, Lipshultz LI and Lamb DJ. Male infertility. In: Strauss JF, Barbieri RL, editors. Yen and Jaffe's reproductive endocrinology: phisiology, pathophysiology and clinical management. Philadelphia: Sanders. 2004. p. 669-71.
World Health Organization. WHO laboratory manual for the examination and processing of human semen. World Health Organization. 2010;5:21.
Gaur DS, Talekar M, Pathak VP. Effect of cigarette smoking on semen quality of infertile men. Singapore Med J. 2007;48(2):119-23.
Gdoura R, Kchaou W, Chaari C, Znazen A, Keskes L, Rebai T, et al. Ureaplasma urealyticum, Ureaplasma parvum, Mycoplasma hominis and Mycoplasma genitalium infections and semen quality of infertile men. BMC Infect Dis. 2007;7:129. http://dx.doi.org/10.1186/1471-2334-7-129
Cooper TG. Sperm maturation in the epididymis: a new look at an old problem. Asian J Androl. 2007;9(4):533-9. http://dx.doi.org/10.1111/j.1745-7262.2007.00285.x
Aitken RJ, Nixon B, Lin M, Koppers AJ, Lee YH, Baker MA. Proteomic changes in mammalian spermatozoa during epididymal maturation. Asian J Androl. 2007;9(4):554-64. http://dx.doi.org/10.1111/j.1745-7262.2007.00280.x
Sipilä P, Pujianto DA, Shariatmadari R, Nikkilä J, Lehtoranta M, Huhtaniemi IT, et al. Differential endocrine regulation of genes enriched in initial segment and distal caput of the mouse epididymis as revealed by genome-wide expression profiling. Biol Reprod. 2006;75(2):240-51. http://dx.doi.org/10.1095/biolreprod.105.047811
Chen J, Wang Y, Xu X, Yu Z, Gui YT, Cai ZM. [Differential expression of ODF1 in human ejaculated spermatozoa and its clinical significance]. Zhonghua Nan Ke Xue. 2009;15(10):891-4. Chinese.
Tarnasky H, Cheng M, Ou Y, Thubdathil JC, Oko R, van der Hoorn FA. Gene trap mutation of murine Outer dense fiber protein-2 gene can result in sperm tail abnormalities in mice with high percentage chimaerism. BMC Developmental Biology. 2010;10:2-11. http://dx.doi.org/10.1186/1471-213X-10-67
Laemmli UK. Cleavage of structural proteins during assembly of the head of the bacteriophage T4. Nature. 1970;227(5259):680-5. http://dx.doi.org/10.1038/227680a0
Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA. 1979;76(9):4350-4. http://dx.doi.org/10.1073/pnas.76.9.4350
Nayernia K, Adham IM, Burkhardt-Göttges E, Neesen J, Rieche M, Wolf S, Sancken U, Kleene K, Engel W. Asthenozoospermia in mice with targeted deletion of the sperm-cystein-rich protein (Smcp) gene. Mol Cell Biol. 2002;22(9):3046-52. http://dx.doi.org/10.1128/MCB.22.9.3046-3052.2002
Brohmann H, Pinnecke S, Hoyer-Fender S. Identification and characterisation of new cDNAs encoding outer dense fiber proteins of rat sperm. J Biol Chem. 1997;272(15):10327-32. http://dx.doi.org/10.1074/jbc.272.15.10327
Hoyer-Fenders, Petersen C, Brohmann H dkk. 1998. Mouse Odf2 cDNAs consist of evolutionary conserved as well as highly variable sequences and encode outer dense fiber proteins of the sperm tail. Mol Reprod Dev. 1998;51:167-75. http://dx.doi.org/10.1002/(SICI)1098-2795(199810)51:2<167::AID-MRD6>3.0.CO;2-O
Petersen C, Füzesi L, Hoyer-Fender S. Outer dense fibre proteins from human sperm tail: molecular cloning and expression analyses of two cDNA transcripts encoding protein of approximately ~70kDa. Mol Hum Reprod. 1999;5(7):627-35. http://dx.doi.org/10.1093/molehr/5.7.627
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