Donmuş boğa spermasında plazma membran sağlamlığının morfolojik bulgularla karşılaştırılması

Hypo-ozmotik ortamda bekletilen spermanın kuyruk kısmında oluşan şişme ve kıvrılmaların hücre membranının fonksiyonel durumunun önemli bir göstergesi olduğu çok sayıda çalışma ile gösterilmiştir. Bu çalışmada rutin sperma analizleri ile elde edilen sonuçların Hypo-ozmotik swelling test (HOST/HOS test) ile karşılaştı-rılması ve bu testlerle birlikte kullanılıp kullanılamayacağı araştırıldı. Çalışmada payet metoduna göre dondurulmuş 6 baş boğanın sperması kullanıldı. Motilite ve spermatozoon morfolojisi (akrozom, baş, orta kısım ve kuyruk anomalileri) ve plazma membran bütünlüğü (HOŞT) 37°C'lik su banyosunda 30 saniyede eritme sonrasında incelendi. Yapılan araştırmada plazma membran bütünlüğüne sahip spermatozoa oranı ile motilite arasında yüksek bir korelasyon olduğu görülmüştür (P0,05) ve orta kısma ait morfolojik bozukluk oranlan (P>0,05) ile membran bütünlüğüne sahip spermatozoa oranları karşılaştırıldığında aralarında herhangi bir korelasyon olmadığı saptanmıştır. Bu çalışmada HOST'nin basit ve uygulanabilir bir test olduğu, rutin semen analizleri ile birlikte kullanılması ile spermanın fonksiyonel bütünlüğü hakkında fikir sahibi olunabileceği sonucuna varıldı.

Anahtar Kelimeler:

spermatozoa, semen koruma, anormallikler, morfoloji, akrozom, dondurarak koruma, analitik yöntemler, boğa, plazma membranı, semen, semen özellikleri

The comparison of plazma membran integrity with morphological characteristics of frozen bull semen

Many studies reporting that swelling of the spermatozoa after exposure to hypo-osmotic condition has been defined as an important sign of functional integrity. The objective of this study was to evaluate the relationship between the results of routine semen analyses and Hypo-osmotic swelling test (HOST/HOS test) and its usability as one of the routine tests. In the present study, straw (0.25 ml) frozen semen from 6 Holstein bulls were used. Post-thaw (37°C for 30s) motility, morphology (acrosome, head, mid piece and tail defect), and plasma membrane integ-rity (HOST) were determined. A significant positive correlation was observed between motility and swollen tail spermatozoa rates (P<0,01) and there was a correlation between tail defect and swollen tail spermatozoa rates (P<0,05) but acrosome, head and mid piece defects were not correlated with swollen tail spermatozoa (P>0,05). In conclusion, the HOS test was simple and may be a useful addition to standard semen analysis for evaluation of sperm functional integrity.

Keywords:

spermatozoa, semen preservation, abnormalities, morphology, acrosome, cryopreservation, analytical methods, bulls, plasma membranes, semen, semen characters,

PDF

___

1.AHMADI, A., SOON-CHYE N.G. The single sperm curling test, a modified hypo-osmotic swelling test, as a potential technique for the selection of viable sperm for intracytoplasmic sperm injection. Fertility and Sterility 1992; 68:346-350.

2.BAHAMONDES, L., FAZANO, F., DE LUCIO, M.A., NEVES, P.A., BOTTOCHER LUIZ, F., LORENZETTI, G.B. Evaluation of human sperm membrane integrity using the watter test and the hypoosmotic test. Andrologia 2001; 33,75-77.

3.BEDFOED, J.M., HOSKINS, D.D. The mammalian spermatozoon: morphology, biochemistry and physiyology in Lamming, G.E. (ed) Marshall’s Physiology of Reproduction. 4. ed. Churchill Livingstone Edinburg London Melbourne and New York 379-568, 1990.

4.BORG, K., COLENBRANDER, B., FAZELI, A., PARLEVLIET, J., MALMGREN, L. Influence of thawing method on motility, plasma membrane integrity and morphology of frozen-thawed stallion spermatozoa. Theriogenology 1997; 48: 531-536.

5.BUCKET, M. W., FARGUHARSON, R. G., LUCKAS, M. J. M., KINGSLAND, C. R., AIRD, I. A., LEWIS-JONES, D. I. The hypoosmotic swelling test in recurrent miscarriage. Fertility and Sterility 1997; 68: 506-509.

6.CALAMERA, J.C., DONCEL, G.F., BRUGO OLMEDO, S., KOLM, P., ACOSTA, A.A. Modified sperm stress test: A simple assay that predicts sperm-related abnormal in-vitro fertilization. Human Reproduction 1998; 13: 2484-2488.

7.CHRISTENSEN, P., BROCKHOFF, P.B., LEHN-JENSEN, H. The relationship between semen quality and the nonreturn rate of bulls. Reprod. Dom. Anim. 1999; 34: 503-507.

8.CORREA, J.A., HEERSCH, G., ZAVOS, P.M. Sperm membrane functional integrity and response of frozen-thawed bovine spermatozoa during the swelling test incubation at varying temperatures. Theriogenology 1997; 47: 715-729.

9.CORREA,J. R., PACE, M.M., ZAVOS, P.M. Relationships among frozen-thawed sperm charecteristics assested via the routine semen analysis, sperm functional tests and fertility of bulls in an artificial insemination program. Theriogenology 1997; 48: 721-731.

10.FOOTE, R.H. Preservation and fertility of spermatozoa. 11.th Intern. Cong. Anim. Reprod. And A. İ., Dublin, 1988; Abstracts Book, 127-134.

11.HAFEZ, E. S. E. Semen evaluation in Hafez, E. S. E. (ed) Reproduction in farm animals. 6. ed. Lea and Fabiger, Philadelphia, 405-423, 1993.

12.İLERI, İ.K., AK, K., PABUÇÇUOĞLU, S., BIRLER, S. Reprodüksiyon ve sun’i Tohumlama. İstanbul Üniv. Vet. Fak. Yayını. Ders Notu No: 23, 2000.

13.JEYENDRAN, R.S., VAN DER VEN, H.H., PEREZ-PELAEZ, M., CRABO. B.G., ZANEVELD, L.J.D. Development of an assay to asses the functional integrity of the human sperm membrane and its relationship to other semen characteristics. J, Reprod. Fert. 1984; 70: 219-228.

14.KUMI-DIAKA, J. Subjecting canine semen to the Hypo-osmotic test. Theriogenology 1993; 39: 1279-1289.

15.LAGARES, M.A., PETZOLDT, R., SIEME, H., KLUG, E. Assessing equine sperm-mebrane integrity. Andrologia 2000; 32. 163-167.

16.LIN, M.-H., MORSHEDI, M., SRISOMBUT, C., NASSAR, A., OEHNINGER, S. Plasma membrane integrity of cryopreserved human sperm: an investigation of the results of the hypoosmotic swelling test, the Water test, and eosin-Y staining. Fertility and Sterility 1998; 70: 6148-155.

17.LIU, Z., FOOTE, R.H. Bull sperm motility and membrane integrity in media varying osmolarity. J. Dairy Sci. 1998; 81: 1868-1873.

18.MING-HUEI, L.,MAHMOOD, M., CHARTCHAI, S., AHMED, N., SERGIO, O. Plazma membrane integrity of cryopreserved human sperm: an investigation of the results of the hypoosmotic swelling test, the water test, and the eosin-Y staining. Fertility and Sterility. 1998; 70: 1148-1155.

19.NEILD, D., CHAVES, G., FLORES, M., MORA, N., BECONI, M., AGÜERO, A. Hypoosmotic test in equine spermatozoa. Theriogenology 1999; 51: 721-727.

20.NEILD, D.M., CHAVES, M.G., FLORES, M., MIRAGAYA, H.M., GONZALES, E., AGÜERO, A. The HOS test and its relationship to fertility in the stallon. Andrologia 2000; 32, 351-355.

21.ROTA, A., PENZO, N., VINCENTI, L., MANTOVANI, R. Hypoosmotic swelling (HOS) as a screening assay for testing in vitro fertility of bovine spermatozoa. Theriogenology 2000; 53: 1415-1420.

22.SÖDERQUIST, L., MADRID-BURY, N., RODRIQUEZ-MARTINEZ, H. Assessment of ram sperm membrane integrity following different thawing procedures. Theriogenology 1991; 48: 1115-1125.

23.VAZQUEZ, J.M., MARTINEZ, E.A., MARTINEZ, P., GARCIA-ARTIGA, C, ROCA, J. Hypoosmotic swelling of boar spermatozoa compared to other methods for analyzing the sperm membrane. Theriogenology 1997; 47: 913-922.