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Detection of the SRY Transcript and Protein in Bovine Ejaculated Spermatozoa

  • Li, Chunjin (College of Animal Science and Veterinary Medicine, and Jilin Provincial Key Laboratory of Animal Embryo Engineering, Jilin University) ;
  • Sun, Yongfeng (College of Animal Science and Veterinary Medicine, and Jilin Provincial Key Laboratory of Animal Embryo Engineering, Jilin University) ;
  • Yi, Kangle (Hunan Institute of Animal and Veterinary Science) ;
  • Li, Chengjiao (College of Animal Science and Veterinary Medicine, and Jilin Provincial Key Laboratory of Animal Embryo Engineering, Jilin University) ;
  • Zhu, Xiaoling (College of Animal Science and Veterinary Medicine, and Jilin Provincial Key Laboratory of Animal Embryo Engineering, Jilin University) ;
  • Chen, Lu (College of Animal Science and Veterinary Medicine, and Jilin Provincial Key Laboratory of Animal Embryo Engineering, Jilin University) ;
  • Zhou, Xu (College of Animal Science and Veterinary Medicine, and Jilin Provincial Key Laboratory of Animal Embryo Engineering, Jilin University)
  • Received : 2011.03.10
  • Accepted : 2011.05.10
  • Published : 2011.10.01

Abstract

The sex-determining region on the Y (SRY) gene is important in mammalian sex determination and differentiation. We report a study of the abundance of SRY gene products in bovine ejaculate. RT-PCR experiments using RNA extracted from bovine spermatozoa with SRY-specific primers yielded a 456 bp product, but the amount of SRY mRNA in sperm was lower than that in the testes (p<0.01). A protein of approximately 27 KDa was detected by western blotting. The SRY transcript was detected in the midpiece of approximately half the spermatozoa by in situ hybridization, and the SRY protein was detected in the heads of half the spermatozoa by immunofluorescence, indicating that SRY mRNA and protein may only be present in Y-bearing spermatozoa. These results suggest that the SRY transcript and protein are present in bovine ejaculated Y-sperm. The roles of the SRY gene in spermatogenesis, sperm motility, and the sperm-oocyte interaction merit further investigation.

Keywords

SRY Gene;Bovine Spermatozoa;In situ Hybridization;Bovine

References

  1. Aquila, S., M. Gentile, E. Middea, S. Catalano and S. Ando. 2004. Autocrine regulation of insulin secretion in human ejaculated spermatozoa. Endocrinology 146(2):552-557. https://doi.org/10.1210/en.2004-1252
  2. Aquila, S., M. Gentile, E. Middea, S. Catalano, C. Morelli, V. Pezzi and S. Ando. 2005. Leptin Secretion by Human Ejaculated Spermatozoa. J. Clin. Endocrinol. Metab. 90(8):4753-4761. https://doi.org/10.1210/jc.2004-2233
  3. Dadoune, J. P., A. Pawlak, M. F. Alfonsi and J. P. Siffroi. 2005. Identification of transcripts by macroarrays, RT-PCR and in situ hybridization in human ejaculate spermatozoa. Mol. Hum. Reprod. 11(2):133-140. https://doi.org/10.1093/molehr/gah137
  4. Dadoune, J. P. 2003. Expression of mammalian spermatozoal nucleoproteins. Microsc. Res. Tech. 61(1):56-75. https://doi.org/10.1002/jemt.10317
  5. Gilbert, I., N. Bissonnette, G. Bossinneault, M. Valee and C. Robert. 2007. A molecilar analysis of the population of mRNA in bovine spermatozoa. Reproduction 133(6):1073-1086. https://doi.org/10.1530/REP-06-0292
  6. Giordano, R., A. R. Magnano, G. Zaccagnini, C. Pittoggi, N. Moscufo, R. Lorenzini and C. Spadafora. 2000. Reverse transcriptase activity in mature spermatozoa of mouse. J. Cell. Biol. 148(6):1107-1113. https://doi.org/10.1083/jcb.148.6.1107
  7. Goodfellow, P. N. and R. Lovell-Badge. 1993. SRY and sex determination in mammals. Annu. Rev. Genet. 27:71-92. https://doi.org/10.1146/annurev.ge.27.120193.000443
  8. Goodwin, L. O., D. S. Karabinus, R. G. Pergolizzi and S. Benoff. 2000. L-type voltage dependent calcium channel alpha-1C subunit mRNA is present in ejaculated human spermatozoa. Mol. Hum. Reprod. 6(2):127-136. https://doi.org/10.1093/molehr/6.2.127
  9. Goodwin, L. O., D. S. Karabinus and R. G. Pergolizzi. 2000. Presence of N-cadherin transcripts in mature spermatozoa. Mol. Hum. Reprod. 6(6):487-497. https://doi.org/10.1093/molehr/6.6.487
  10. Gur, Y. and H. Breitbart. 2006. Mammalian sperm translate nuclear-encoded proteins by mitochondrial-type ribosomes. Genes Dev. 20(4):411-416. https://doi.org/10.1101/gad.367606
  11. Hacker, A., B. Capel, P. N. Goodfellow and R. Lovell-Badge. 1995. Expression of SRY, the mouse sex determining gene. Development 121(6):1603-1614.
  12. Hanley, N. A., D. M. Hagan, M. Clement-Jones, S. G. Ball, T. Strachan, L. Salsa-Cortes, K. McElreavey, S. Lindsay, S. Robson, P. Bullen, H. Ostrer and D. I. Wilson. 2000. SRY, SOX9 and DAX1 expression patterns during human sex determination and gonadal development. Mech. Dev. 91(1-2):403-407. https://doi.org/10.1016/S0925-4773(99)00307-X
  13. Koopman, P., J. Gubbay, N. Vivian, P. Goodfellow and R. Lovell-Badge. 1991. Male development of chromosomally female mice transgenic for Sry. Nature 351(6322):117-121. https://doi.org/10.1038/351117a0
  14. Lambard, S., I. Galeraud-Denis, G. Martin, R. Levy, A. Chocat and S. Carreau. 2004. Analysis and significance of mRNA in human ejaculated sperm from normozoospermic donors: relationship to sperm motility and capacitation. Mol. Hum. Reprod. 10(7):535-541. https://doi.org/10.1093/molehr/gah064
  15. Lambard, S., I. Galeraud-Denis, P. T. Saunders and S. Carreau. 2004. Human immature germ cells and ejaculated spermatozoa contain aromatase and oestrogen receptors. J. Mol. Endocrinol. 32(1):279-289. https://doi.org/10.1677/jme.0.0320279
  16. Li, C. J., C. J. Li, X. L. Zhun, C. Q. Wang, Z. Liu, W. H. Li and C. Lu. The expression and putative role of brain-derived neurotrophic factor and its receptor in bovine sperm. (unpublished)
  17. Lin, Y. M., C. W. Chen, H. S. Sun, S. J. Tsai, J. S. Nan and P. L. Kuo. 2002. Presence of DAZL transcript and protein in mature human spermatozoa. Fertil. Steril. 77(3):626-629. https://doi.org/10.1016/S0015-0282(01)03226-5
  18. Modi, D., P. Berde and D. Bhartiya. 2003. Down syndrome: a study of chromosomal mosaicism. Reprod. Biomed. Online. 6(4):499-503. https://doi.org/10.1016/S1472-6483(10)62174-8
  19. Modi, D., S. Sane and D. Bhartiya. 2003. Accelerated germ cell apoptosis in sex chromosome aneuploid fetal human gonads. Mol. Hum. Reprod. 9(4):219-225. https://doi.org/10.1093/molehr/gag031
  20. Modi, D., C. Shah, G. Sachdeva, S. Gadka, D. Bhartiya and C. Puri. 2005. Ontogeny and cellular location of SRY transcripts in the human testes and its detection in spermatozoa. Reproduction 130(5):603-613. https://doi.org/10.1530/rep.1.00413
  21. Ostermeier, G. C., D. J. Dix, D. Miller, P. Khatri and S. A. Krawetz. 2002. Spermatozoal RNA profiles of normal fertile men. Lancet 360(9335):772-777. https://doi.org/10.1016/S0140-6736(02)09899-9
  22. Ostermeier, G. C., D. Miller, J. D. Huntriss, M. P. Diamond and S. A. Krawetz. 2004. Reproductive biology: delivering spermatozoan RNA to the oocyte. Nature 429(6988):154.
  23. Parrish, J. J., J. L. Susko-Parrish, M. L. Leibfried-Rutledge, E. S. Critser, W. H. Eyestone and N. L. First. 1986. Bovine in vitro fertilization with frozen-thawed semen. Theriogenology 25(4):591-600. https://doi.org/10.1016/0093-691X(86)90143-3
  24. Parsons, R. 2004. Human cancer, PTEN and the PI-3 kinase pathway. Semin. Cell Dev. Biol. 15(2):171-176. https://doi.org/10.1016/j.semcdb.2003.12.021
  25. Shah, C. A., D. Modi, G. Sachdeva, S. Gadkar, S. D'Souza and C. P. Puri. 2005. N terminal region of progesterone receptor B isoform in human spermatozoa. Int. J. Androl. 28(6):360-371. https://doi.org/10.1111/j.1365-2605.2005.00566.x
  26. Steger, K. 2003. Perspectives in the diagnosis of testicular biopsies using molecular biological techniques. Andrologia 35(3):183.
  27. Zuccarello, D., A. Garolla, A. Ferlin, M. Menegazzo, L. D. Toni, M. Carraro, C. Veronese and C. Foresta. 2009. Androgen receptor is expressed in both X- and Y-carrier human spermatozoa. Fertil. Steril. 91(1):193-200. https://doi.org/10.1016/j.fertnstert.2007.11.040

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