Journal of Embryo Transfer (한국수정란이식학회지)

The Korean Society of Embryo Transfer (한국수정란이식학회)
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Assessment of Jeju Horse Semen using Computer-Assisted Sperm Analysis (CASA)

Computer-Assisted Sperm Analysis(CASA)를 이용한 제주마 정액의 운동성 평가

  • Kang T.Y. (Department of Veterinary Medicine, Cheju National University) ;
  • Kang M.S. (Major of Animal Science and Biotechnology, Cheju National University)
  • 강태영 (제주대학교 수의학과) ;
  • 강민수 (제주대학교 생명자원과학대학 생물산업학부)
  • Published : 2006.03.01
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Abstract

The objective of the study was to assess the general characteristics and motility characteristics with Computer Assisted Sperm Analyzer (CASA) system in Jeju horse semen. Semen was collected from 5 fertile Jeju horse by use of a Missouri type artificial vagina. Semen volume and pH were recorded, and sperm concentration was determined with a hematocytometer and motional characteristics of sperm were analysed by CASA. The viability and morphological abnormalities were assessed by a vital staining. The average volume of ejaculates was 42.5 ml and the average of sperm concentration was $198.5x10^6/m1$. The motional characteristics in Jeju horse semen was showed $70.4{\pm}28.7{\mu}m/s\;for\;VAP,\;69.6{\pm}28.9{\mu}m/s\;for\;VSL,\;94.1{\pm}30.0{\mu}m/s\;fo\;VCL,\;2.3{\pm}0.7{\mu}m/s\;for\;ALH,\;7.6{\pm}1.1Hz\;for\;BCF,\;99.1{\pm}1.2%\;for\;STR,\;and\;77.1{\pm}12.7%\;for\;LIN$. The percentage of sperm with abnormal head, midpiece and tail was 4.2%, 20.6%, 4.6% respectively.

본 연구는 제주도 축산진흥원과 농촌진흥청 난지농업연구소에서 사육되고 있는 제주마 5두를 사용하여 인공질을 이용, 정액을 채취하여 정액의 일반성상 및 CASA를 이용하여 정자의 운동성과 생사 염색으로 기형률을 조사하였다. 제주마 정액의 일반 특성으로 총 정액량은 평균 42.5ml이었으며 평균 pH는 7.3, 평균 정자농도는 평균 $198.5X10^6/m1$ 이었다. 정자의 운동성은 $64.3{\pm}23.2%$이며, motility parameters 별로 VAP $70.4{\pm}28.7{\mu}m/s,\;VSL\;69.6{\pm}28.9{\mu}m/s,\;VCL\;94.1{\pm}30.0{\mu}m/s,\;ALH\;2.3{\pm}0.7{\mu}m/s,\;BCF\;7.6{\pm}1.1Hz,\;STR\;99.1{\pm}1.2%,\;LIN\;77.1{\pm}12.7%$로 나타났다. 정자 부위별 기형률은 두부가 평균 4 %, 경부가 평균 20 %, 미부가 평균 4 %로 나타났다. 이 연구를 통해 천연기념물로 지정되어 있는 제주마의 유전자원 보존과 증식을 위한 인공수정기술의 실용화를 위한 기초 자료를 제공하고자 한다.

Keywords

References

  1. Agarwal A, Sharma RK and Nelson DR. 2003. New semen quality scores developed by principal component analysis of semen characteristics. J. Androl., 24:343-352 https://doi.org/10.1002/j.1939-4640.2003.tb02681.x
  2. Armann RP. 1989. Can the fertility potential of a seminal sample be predicted accurately? J. Androl., 10:89-98 https://doi.org/10.1002/j.1939-4640.1989.tb00066.x
  3. Armant DR and Ellis MA. 1995. Improved accuracy of sperm motility assessment using a modified Micro-Cell sperm counting chamber. Fertil. Steril., 63:1128-1130 https://doi.org/10.1016/S0015-0282(16)57562-1
  4. Auger J, Leonce S, Jouannet P and Ronot X. 1993. Flow cytometric sorting of living, highly motile human spermatozoa based on evaluation of their mitochondrial activity. J. Histochem. Cytochem., 41:1247-1251 https://doi.org/10.1177/41.8.8331289
  5. Bielanski W, Dudek E, Bittmar A and Kosiniak K. 1982. Some characteristis of common abnormal forms of spermatozoa in highly fertile stallions. J. Reprod. Fert. Suppl., 32:21-26
  6. Budworth PR, Amann RP and Chapman PL. 1988. Relationships between computerized measurements of motion of frozen-thawed bull spermatozoa and fertility. J. Androl., 9:41-54 https://doi.org/10.1002/j.1939-4640.1988.tb01007.x
  7. Chan SY, Wang C, Song BL, Lo T, Leung A, Tsoi WL and Leung J. 1989. Computer-assisted image analysis of sperm concentration in human semen before and after swim-up separation: Comparison with assessment by haemocytometer. Int. J. Androl., 12:339-345 https://doi.org/10.1111/j.1365-2605.1989.tb01322.x
  8. Coetzee K, Kruger TF and Lombard CJ. 1999. Repeatability and variance analysis on multiple computer-assisted (IVOS) sperm morphology readings. Andrologia, 31: 163-168 https://doi.org/10.1046/j.1439-0272.1999.00257.x
  9. Davis RO and Siemers RJ. 1995. Derivation and reliability of kinematic measures of sperm motion. Reprod. Fertil. Dev., 7:857-869 https://doi.org/10.1071/RD9950857
  10. Davis RO, Drobnis EZ and Overstreet JW. 1995. Application of multivariate cluster, discriminate function, and stepwise regression analyses to variable selection and predictive modeling of sperm cryosurvival. Fertil. Steril., 63:1051-1057 https://doi.org/10.1016/S0015-0282(16)57547-5
  11. Donnelly ET, Lewis SE, McNally JA and Thompson W. 1998. In vitro fertilization and pregnancy rates: the influence of sperm motility and morphology on IVF outcome. Fertil. Steril., 70: 305-314 https://doi.org/10.1016/S0015-0282(98)00146-0
  12. Dowsett KF and Pattie WA. 1982. Characteristics and fertility of stallion semen. J. Reprod. Fert. Suppl., 44:77-86
  13. Dunphy BC, Kay R, Barratt CL and Cooke ID. 1989. Quality control during the conventional analysis of semen, an essential exercise. J. Androl., 10:378-385 https://doi.org/10.1002/j.1939-4640.1989.tb00124.x
  14. Farrell PB, Foote RH, McArdle MM, Trouern-Trend VL and Tardif AL. 1996. Media and dilution procedures tested to minimize handling effects on human, rabbit, and bull sperm for computer-assisted sperm analysis (CASA). J. Androl., 17:293-300
  15. Fernandes PA, McCoshen JA, Cheang M, Kredentser JV and Wodzicki AM. 1990. Quantitative analysis of the effect of freezing on donor sperm motion kinetics. Fertil. Steril., 54:322-327 https://doi.org/10.1016/S0015-0282(16)53711-X
  16. Hirano Y, Shibahara H, Obara H; Suzuki T, Takamizawa S, Yamaguchi C, Tsunoda H and Sato I. 2001. Relationships between sperm motility characteristics assessed by the computer-aided sperm analysis (CASA) and fertilization rates in vitro. J. Assist. Reprod. Genet., 18:213-218
  17. Holt W, Watson P, Curry M and Holt C. 1994. Reproducibility of computer-aided semen analysis: comparison of five different systems used in a practical workshop. Fertil. Steril., 62:1277-1282 https://doi.org/10.1016/S0015-0282(16)57201-X
  18. Iguer-Ouada M and Verstegen JP. 2001. Evaluation of the 'Hamilton Thorn computer-based automated system' for dog semen analysis. Theriogenology, 55:733-749 https://doi.org/10.1016/S0093-691X(01)00440-X
  19. Imade GE, Towobola OA, Sagay AS and Otubu JA. 1993. Discrepancies in sperm count using improved Neubauer, Makler, and Horwells counting chambers. Arch. Androl., 31:17-22 https://doi.org/10.3109/01485019308988375
  20. Jequier AM and Ukombe EB. 1983. Errors inherent in the performance of a routine semen analysis. Br. J. Urol., 55:434-436 https://doi.org/10.1111/j.1464-410X.1983.tb03339.x
  21. Jeulin C, Lewin LM, Chevrier C and Schoevaert Brossault TD. 1996. Changes in flagella movement of rat spermatozoa along the length of the epididymis : Manual and computer-aided image analysis. Cell. Motil. Cystoskeleton, 35:147-161 https://doi.org/10.1002/(SICI)1097-0169(1996)35:2<147::AID-CM7>3.0.CO;2-7
  22. Johnson JE, Boone WR and Blackhurst DW. 1996. Manual versus computer-automated semen analyses. Part II. Determination of the working range of a computer-automated semen analyzer. Fertil. Steril., 65:156-159 https://doi.org/10.1016/S0015-0282(16)58044-3
  23. Kruger TF, Acosta AA, Simmons KF, Swanson RJ, Matta JF and Oehninger S. 1988. Predictive value of abnormal sperm morphology in in vitro fertilization. Fertil. Steril., 49: 112-117 https://doi.org/10.1016/S0015-0282(16)59660-5
  24. Lackey BR, Boone WR, Gray SL and Henricks DM. 1998. Computer-assisted sperm motion analysis of bovine sperm treated with insulin-like growth factor I and II: implications as motility regulators and chemokinetic factors. Arch. Androl., 41:115-125 https://doi.org/10.3109/01485019808987953
  25. Magistrini M, Chanteloube P and Palmer E. 1987. Influence of season and frequency of ejaculation on production of stallion semen for freezing. J. Reprod. Fertil. Suppl., 35:127-133
  26. Marshburn PB, McIntire D, Carr BR and Byrd W. 1992. Spermatozoal characteristics from fresh and frozen donor semen and their correlation with fertility outcome after intrauterine insemination. Fertil. Steril., 58: 179-188 https://doi.org/10.1016/S0015-0282(16)55157-7
  27. McDonnell SM and Hize AL. 2005. Aversive conditioning of periodic spontaneous erection adversely affects sexual behavior and semen in stallions. Anim. Reprod. Sci., 89:77-92 https://doi.org/10.1016/j.anireprosci.2005.06.016
  28. Purvis K, Tollefsrud A and Rui H. 1989. Correlates of human sperm motility assessed by laser Doppler spectroscopy. Int. J. Androl., 12:10-16 https://doi.org/10.1111/j.1365-2605.1989.tb01280.x
  29. Pusch HH, Leitinger S, Winter R and Urdl W. 1986. Evaluation of sperm motility by laser-Doppler-spectroscopy in an in-vitro fertilization programme. Hum. Reprod., 1:233-235 https://doi.org/10.1093/oxfordjournals.humrep.a136391
  30. Quintero-Moreno A, Miro J, Teresa Rigau A and Rodriguez-Gil JE. 2003. Identification of sperm subpopulations with specific motility characteristics in stallion ejaculates. Theriogenology, 59: 1973-1990 https://doi.org/10.1016/S0093-691X(02)01297-9
  31. Rashid MRZ, Fishel SB and Thornton S. 1998. The predictive value of the zona free hamster egg penetration test in relation to in vitro fertilization at various insemination concentrations. Hum. Reprod., 13:624-629 https://doi.org/10.1093/humrep/13.3.624
  32. Squires EL, Pickett BW, Graham JK, Vanderwall DK, McCue PM and Bruemmer JE. 1999. Cooled and frozen stallion semen. Anim. Reprod. Biotech. Lab. Colorado State Univ. Bulletin No. 09
  33. Thurston LM, Watson PF, Mileham AJ and Holt WV. 2001. Morphologically distinct sperm subpopulations defined by Fourier shape descriptors in fresh ejaculates correlate with variation in boar semen quality following cryopreservation. J. Androl., 22:382-394
  34. Vetter CM, Miller JE, Crawford LM, Armstrong MJ, Clair JH, Conner MW, Wise LD and Skopek TR. 1998. Comparison of motility and membrane integrity to assess rat sperm viability. Reprod. Toxicol., 12:105-114 https://doi.org/10.1016/S0890-6238(97)00155-X
  35. 박남건, 오운용, 이성수, 오창언, 강수원, 고서봉, 강민수, 김희석. 1995. 제주재래마 정액의 동결 보존에 관한 연구. 농업논문집, 37:459-463
  36. 양보석, 강승률, 이성수, 조인철, 검영훈, 전창익, 정진관, 2001. 제주마 정액의 일반성상에 관한 연구. 한국수정란이식학회지, 16: 127-131
  37. 오운용, 박남건, 김영훈, 이성수, 김희석, 김중계, 신원집. 1994. 제주재래마 정액의 일반성상에 관한 조사연구. 농업논문집, 36:552-557