Effects of cryopreservation on the acrosomal status of stallion spermatozoa.

S. J. Bedford, D. D. Varner, Stuart A Meyers

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

The effects of cryopreservation on the acrosomal status of equine spermatozoa were investigated. Ejaculates (n=10) from six stallions were processed fresh, after cooled storage at 4-6 degrees C for 24 h in either a milk-based or lactose-EDTA freezing extender and after freeze-thawing in lactose-EDTA extender in liquid nitrogen at either 5 x 10(7) or 2 x 10(8) spermatozoa ml(-1). All samples were incubated in TALP-TEST for 2 h at 39 degrees C in 5% CO2. Subsamples were challenged with calcium ionophore A23187 for 10 min. The acrosomal status of the spermatozoa was evaluated by staining the spermatozoa with FITC-conjugated Pisum sativum agglutinin, with ethidium homodimer as a nuclear counterstain. Sperm cell viability was assessed with Hoechst 33258. The data were analysed by a general linear model ANOVA (P < 0.05). Treatment with calcium ionophore did not affect the percentage of acrosome reactions. The samples containing 5 x 10(7) and 2 x 10(8) spermatozoa ml(-1) that were frozen in lactose-EDTA extender in liquid nitrogen had higher percentages of spermatozoa in intermediate categories of acrosomal staining than did any other treatment. The percentage of acrosome-reacted cells was also higher overall for these samples. The percentage of viable cells was lowest for the sperm samples frozen in lactose-EDTA extender, and lower in semen stored in either a milk-based or lactose-EDTA freezing extender than in fresh semen. In conclusion, freeze-thawing resulted in a high percentage of acrosomal changes and a significant decrease in sperm viability.

Original languageEnglish (US)
Pages (from-to)133-140
Number of pages8
JournalJournal of reproduction and fertility. Supplement
Issue number56
StatePublished - 2000

Fingerprint

Cryopreservation
Spermatozoa
Lactose
Edetic Acid
Calcium Ionophores
Semen
Freezing
Milk
Nitrogen
Staining and Labeling
Bisbenzimidazole
Acrosome Reaction
Acrosome
Fluorescein-5-isothiocyanate
Agglutinins
Peas
Calcimycin
Horses
Linear Models
Cell Survival

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Effects of cryopreservation on the acrosomal status of stallion spermatozoa. / Bedford, S. J.; Varner, D. D.; Meyers, Stuart A.

In: Journal of reproduction and fertility. Supplement, No. 56, 2000, p. 133-140.

Research output: Contribution to journalArticle

@article{2cfd13a444a647269fa273d3f1aaeb02,
title = "Effects of cryopreservation on the acrosomal status of stallion spermatozoa.",
abstract = "The effects of cryopreservation on the acrosomal status of equine spermatozoa were investigated. Ejaculates (n=10) from six stallions were processed fresh, after cooled storage at 4-6 degrees C for 24 h in either a milk-based or lactose-EDTA freezing extender and after freeze-thawing in lactose-EDTA extender in liquid nitrogen at either 5 x 10(7) or 2 x 10(8) spermatozoa ml(-1). All samples were incubated in TALP-TEST for 2 h at 39 degrees C in 5{\%} CO2. Subsamples were challenged with calcium ionophore A23187 for 10 min. The acrosomal status of the spermatozoa was evaluated by staining the spermatozoa with FITC-conjugated Pisum sativum agglutinin, with ethidium homodimer as a nuclear counterstain. Sperm cell viability was assessed with Hoechst 33258. The data were analysed by a general linear model ANOVA (P < 0.05). Treatment with calcium ionophore did not affect the percentage of acrosome reactions. The samples containing 5 x 10(7) and 2 x 10(8) spermatozoa ml(-1) that were frozen in lactose-EDTA extender in liquid nitrogen had higher percentages of spermatozoa in intermediate categories of acrosomal staining than did any other treatment. The percentage of acrosome-reacted cells was also higher overall for these samples. The percentage of viable cells was lowest for the sperm samples frozen in lactose-EDTA extender, and lower in semen stored in either a milk-based or lactose-EDTA freezing extender than in fresh semen. In conclusion, freeze-thawing resulted in a high percentage of acrosomal changes and a significant decrease in sperm viability.",
author = "Bedford, {S. J.} and Varner, {D. D.} and Meyers, {Stuart A}",
year = "2000",
language = "English (US)",
pages = "133--140",
journal = "Journal of reproduction and fertility. Supplement",
issn = "0449-3087",
publisher = "Society for Reproduction and Fertility",
number = "56",

}

TY - JOUR

T1 - Effects of cryopreservation on the acrosomal status of stallion spermatozoa.

AU - Bedford, S. J.

AU - Varner, D. D.

AU - Meyers, Stuart A

PY - 2000

Y1 - 2000

N2 - The effects of cryopreservation on the acrosomal status of equine spermatozoa were investigated. Ejaculates (n=10) from six stallions were processed fresh, after cooled storage at 4-6 degrees C for 24 h in either a milk-based or lactose-EDTA freezing extender and after freeze-thawing in lactose-EDTA extender in liquid nitrogen at either 5 x 10(7) or 2 x 10(8) spermatozoa ml(-1). All samples were incubated in TALP-TEST for 2 h at 39 degrees C in 5% CO2. Subsamples were challenged with calcium ionophore A23187 for 10 min. The acrosomal status of the spermatozoa was evaluated by staining the spermatozoa with FITC-conjugated Pisum sativum agglutinin, with ethidium homodimer as a nuclear counterstain. Sperm cell viability was assessed with Hoechst 33258. The data were analysed by a general linear model ANOVA (P < 0.05). Treatment with calcium ionophore did not affect the percentage of acrosome reactions. The samples containing 5 x 10(7) and 2 x 10(8) spermatozoa ml(-1) that were frozen in lactose-EDTA extender in liquid nitrogen had higher percentages of spermatozoa in intermediate categories of acrosomal staining than did any other treatment. The percentage of acrosome-reacted cells was also higher overall for these samples. The percentage of viable cells was lowest for the sperm samples frozen in lactose-EDTA extender, and lower in semen stored in either a milk-based or lactose-EDTA freezing extender than in fresh semen. In conclusion, freeze-thawing resulted in a high percentage of acrosomal changes and a significant decrease in sperm viability.

AB - The effects of cryopreservation on the acrosomal status of equine spermatozoa were investigated. Ejaculates (n=10) from six stallions were processed fresh, after cooled storage at 4-6 degrees C for 24 h in either a milk-based or lactose-EDTA freezing extender and after freeze-thawing in lactose-EDTA extender in liquid nitrogen at either 5 x 10(7) or 2 x 10(8) spermatozoa ml(-1). All samples were incubated in TALP-TEST for 2 h at 39 degrees C in 5% CO2. Subsamples were challenged with calcium ionophore A23187 for 10 min. The acrosomal status of the spermatozoa was evaluated by staining the spermatozoa with FITC-conjugated Pisum sativum agglutinin, with ethidium homodimer as a nuclear counterstain. Sperm cell viability was assessed with Hoechst 33258. The data were analysed by a general linear model ANOVA (P < 0.05). Treatment with calcium ionophore did not affect the percentage of acrosome reactions. The samples containing 5 x 10(7) and 2 x 10(8) spermatozoa ml(-1) that were frozen in lactose-EDTA extender in liquid nitrogen had higher percentages of spermatozoa in intermediate categories of acrosomal staining than did any other treatment. The percentage of acrosome-reacted cells was also higher overall for these samples. The percentage of viable cells was lowest for the sperm samples frozen in lactose-EDTA extender, and lower in semen stored in either a milk-based or lactose-EDTA freezing extender than in fresh semen. In conclusion, freeze-thawing resulted in a high percentage of acrosomal changes and a significant decrease in sperm viability.

UR - http://www.scopus.com/inward/record.url?scp=0000252682&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0000252682&partnerID=8YFLogxK

M3 - Article

C2 - 20681125

AN - SCOPUS:0000252682

SP - 133

EP - 140

JO - Journal of reproduction and fertility. Supplement

JF - Journal of reproduction and fertility. Supplement

SN - 0449-3087

IS - 56

ER -