Adaptation of the Hypoosmotic Swelling Test to Evaluate Membrane Integrity of Boar Spermatozoa

  • 발행 : 2007.06.30

초록

The objective of this study was to establish the optimal conditions for hypoosmotic swelling (HOS) test to assess the functional integrity of the membranes of boar fresh or frozen/thawed spermatozoa. When pooled semen sample was incubated for 30 min at $37^{\circ}C$ with different test solution of varied osmolarity, the highest percentage of HOS positive spermatozoa was observed in a 150 mOsmol fructose/Na-citrate solution (33.6%). Incubation time did not affect significantly the score of HOS positive spermatozoa observed in a 150 mOsmol fructose/Na-citrate solution at $37^{\circ}C$, but the osmolarity affected the score of HOS positive spermatozoa under the same condition above. Fresh semen was significantly better than frozen/thawed semen in semen parameters evaluated such as motility, viability, membrane integrity and lipid peroxidation (p<005). In the relationships of sperm parameters, motility vs viability, motility vs membrane integrity and viability vs membrane integrity were positively correlated ($0.82{\sim}0.94$) but lipid peroxidation vs other estimated factors was negatively correlated ($- 0.90{\sim}- 0.98$). Among the evaluation methods, motility vs Viability, motility vs membrane integrity and lipid peroxidation vs other estimated factors were significantly correlated (p<0.05). These results of this. study indicate that the optimal condition of HOST in boar spermatozoa is a 150 mOsmol fructose/Na-citrate solution for 30 min incubation at $37^{\circ}C$ and HOST can substitute the examination of motility, viability and lipid peroxidation.

키워드

참고문헌

  1. Caiza de la Cueva FI, Rigau T, Bonet S, Miro J, Briz M, Rodriquez-Gil JE (1997): Subjecting horse spermatozoa to hypoosmotic incubation: effects of ouabain. Theriogenology 47:765-784 https://doi.org/10.1016/S0093-691X(97)00033-2
  2. Cerolini S, Maldjian A, Surai P, Noble R (2000): Viability, susceptibility to peroxidation and fatty acid composition of boar semen during liquid storage. Anim Reprod Sci 58:99-111 https://doi.org/10.1016/S0378-4320(99)00035-4
  3. Correa JR, Zavos PM (1994): The hypoosmotic swelling test: Its employment as an assay to evaluate the functional integrity of the frozen-thawed bovine sperm membrane. Theriogenology 42:351-360 https://doi.org/10.1016/0093-691X(94)90280-1
  4. Francisco I, Caiza De La Cueva, Teresa Rigau, Rosa Pujol, Jesus Piedrafita, Rodriguez-Gil JE (1997): Resistance to hyperosmotic stress in boar spermatozoa: the role of the ionic pumps and the relationship with cryosurvival. Animal Reprod Sci 48:301-315 https://doi.org/10.1016/S0378-4320(97)00061-4
  5. Gadea J, Matas C (2000): Sperm factors related to in vitro penetrationof porcine oocytes. Theriogenology 54:1343-1357 https://doi.org/10.1016/S0093-691X(00)00458-1
  6. Iguer-ouada M, Verstegen J (2001): Validation of the sperm Quality Analyzer (SQA) for dog sperm analysis. Theriogenology 55:1143-1158 https://doi.org/10.1016/S0093-691X(01)00473-3
  7. Jeyendran RS, Van der Ven HH, Perez-Pelaez M, Crabo BG, Zaneveld LJD (1984): Development of an assay to assess the functional integrity of the human sperm membrane and its relationship to other semen characteristics. J Reprod Fertil 70:219-228 https://doi.org/10.1530/jrf.0.0700219
  8. Nie GJ, Wenzel JGW (2000): Adaptation of the hypoosmotic swelling test to assess functional integrity of stallion spermatozoal plasma membranes. Theriogenology 55:1005-1018 https://doi.org/10.1016/S0093-691X(01)00461-7
  9. Perez-Llano B, Lorenzo JL, Yenes P, Trejo A, García-Casado P (2001): A short hypoosmotic swelling test for the prediction of boar sperm fertility Theriogenology 56:387-398 https://doi.org/10.1016/S0093-691X(01)00571-4
  10. Rodriquez-Gil JE, Rigau T (1996): Effects of ouabain in the response to osmotic changes in dog and boar spermatozoa. Theriogenology 45:873-888 https://doi.org/10.1016/0093-691X(96)00016-7
  11. Vazquez JM, Martinez EA, Martinez P, Garcia-Artiga C, Roca J (1997): Hypoosmotic swelling of boar spermatozoa compared to other methods for analyzing the sperm membrane. Theriogenology 913-922
  12. Zaneveld LJD, Jeyendren RS, Krajeski P, Coetzee K, Kruger TF, Lombard LJ (1990): Hopoosmotic swelling test. In: Acosta AA, Swanson RJ, Ackerman SB, Kruger TF, Van Zyl JA, Menkveld R (eds), Human spermatozoa in assisted reproduction. Williams and Wilkins, pp 223-227
  13. Zavos PM, Correa GM, Zarmakoupis PN (1994): Improvements and shortterm viability of mouse epididymal spermatozoa recovered through the Sperm Prep filtration method. Theriogenology 42:1036-1042