Register      Login
Reproduction, Fertility and Development Reproduction, Fertility and Development Society
Vertebrate reproductive science and technology
Shopping Cart: ( empty )
You are here: Home > Journals > RD > RD05003
RESEARCH ARTICLE
Contents Vol 17(4)

Effects of hypothermic liquid storage and cryopreservation on basal and induced plasma membrane phospholipid disorder and acrosome exocytosis in boar spermatozoa*

H. D. Guthrie A B and G. R. Welch A
+ Author Affiliations
- Author Affiliations

A Biotechnology and Germplasm Laboratory, Agricultural Research Service, U. S. Department of Agriculture, Beltsville, MD 20705, USA.

B Corresponding author. Email: dave@anri.barc.usda.gov

Reproduction, Fertility and Development 17(4) 467-477 https://doi.org/10.1071/RD05003
Submitted: 4 January 2005  Accepted: 27 February 2005   Published: 12 April 2005

Abstract

Flow cytometry was utilised to determine whether short-term (Day 1) or long-term hypothermic liquid storage (Day 5), or cryopreservation of boar spermatozoa (1) caused changes in plasma membrane phospholipid disorder (MPLD) and acrosome exocytosis (AE), indicative of an advanced stage of capacitation or acrosome status, and (2) facilitated or inhibited the induction of capacitation and the acrosome reaction. Merocyanine with Yo-Pro-1 and peanut agglutinin–fluorescein isothiocyanate with propidium iodide were used to identify MPLD and AE, respectively, in viable spermatozoa. The incidence of basal sperm MPLD and AE in fresh semen was very low (1.1 and 2.2%, respectively) and was increased (P < 0.05) only a small amount in Day 5 and cryopreserved semen (3–8%). Compared to no bicarbonate, incubation with bicarbonate increased MPLD, but the response was greatest (P < 0.05) in fresh sperm (52.3%) compared with Day 1 (36.6%), Day 5 (13.9%) and cryopreserved sperm (13.6%). Incubation with calcium ionophore A23187 increased AE in spermatozoa, but the response was less (P < 0.05) for fresh (34%) and cryopreserved (27%) semen than for Day 1 (45%) and Day 5 (57%) semen. In summary, hypothermic liquid storage and cryopreservation of boar spermatozoa did not advance capacitation or acrosome status in viable spermatozoa, but did alter their responses to induction of capacitation and the acrosome reaction.

Extra keywords: peanut agglutinin–fluorescein isothiocyanate, membrane integrity, merocyanine, motility.


References

Almlid, T. , and Johnson, L. A. (1988). Effects of glycerol concentration, equilibration time, and temperature of glycerol addition on post-thaw viability boar spermatozoa frozen in straws. J. Anim. Sci. 66, 2899–2905.
| PubMed | Colenbrander B., Gadella B., Feitsma H., and Woelders H. (2000). Semen quality assessment, present and future. In ‘Boar Semen Preservation IV’. (Eds L. A. Johnson and H. D. Guthrie.) pp. 35–41. (Allen Press, Inc.: Lawrence, KS, USA.)

Colenbrander, B. , Gadella, B. M. , and Stout, T. A. E. (2003). The predictive value of semen analysis in the evaluation of stallion fertility. Reprod. Dom. Anim. 38, 305–311.
| Crossref | GoogleScholarGoogle Scholar | Johnson L. A. (1985). Fertility results using frozen boar spermatozoa: 1970–1985. In ‘Deep Freezing of Boar Semen’. (Eds L. A. Johnson and K. Larsson.) pp. 199–222. (Swedish University of Agricultural Science: Uppsala, Sweden.)

Johnson, L. A. , Weitze, K. F. , Fiser, P. , and Maxwell, W. M. C. (2000). Storage of boar semen. Anim. Reprod. Sci. 62, 143–172.
| Crossref | GoogleScholarGoogle Scholar | PubMed | Little R. C., Milliken G. A., Stroup W. W., and Wolfinger R. D. (1996). ‘SAS System for Mixed Models.’ (SAS Institute Inc.: Cary, NC, USA.)

Mattioli, M. , Barboni, B. , Lucidi, P. , and Seren, E. (1996). Identification of capacitation in boar spermatozoa by chlortetracycline staining. Theriogenology 45, 373–381.
| Crossref | GoogleScholarGoogle Scholar | Tsien R. Y. (1989). Fluorescent indicators of ion concentrations. In ‘Fluorescence Microscopy of Living Cells in Culture. Part 6B. Quantitative Fluorescence Microscopy—Imaging and Spectroscopy. Methods in Cell Biology. Vol 30’. (Eds D. L. Taylor and Y. L. Wang.) pp. 127–156. (Academic Press: New York, USA.)

Watson, P. F. (1995). Recent developments and concepts in the cryopreservation of spermatozoa and the assessment of their post-thawing function. Reprod. Fertil. Dev. 7, 871–891.
| PubMed | Yanagimachi R. (1994). Mammalian fertilization. In ‘The Physiology of Reproduction’. 2nd edn. (Eds E. Knobil and J. D. Neill.) pp. 189–317. (Raven Press Ltd: New York, USA.)




* Mention of a trade name or proprietary product does not constitute a guarantee or warranty by the USDA and does not imply approval to the exclusion of others not mentioned.