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Heparin and dermatan sulphate induced capacitation of frozen-thawed bull spermatozoa measured by merocyanine-540

Published online by Cambridge University Press:  01 August 2007

A.-S. Bergqvist ,
J. Ballester ,
A. Johannisson ,
N. Lundeheim  and
H. Rodríguez-Martínez
A.-S. Bergqvist*
Affiliation:
Division of Comparative Reproduction, Obstetrics and Udder Health, Department of Clinical Sciences, Faculty of Veterinary Medicine and Animal Science, Swedish University of Agricultural Sciences (SLU), Uppsala, Sweden.
J. Ballester
Affiliation:
Department of Anatomy and Physiology, Faculty of Veterinary Medicine and Animal Science, SLU, Uppsala, Sweden.
A. Johannisson
Affiliation:
Department of Anatomy and Physiology, Faculty of Veterinary Medicine and Animal Science, SLU, Uppsala, Sweden.
N. Lundeheim
Affiliation:
Department of Animal Breeding and Genetics, Faculty of Veterinary Medicine and Animal Science, SLU, Uppsala, Sweden.
H. Rodríguez-Martínez
Affiliation:
Division of Comparative Reproduction, Obstetrics and Udder Health, Department of Clinical Sciences, Faculty of Veterinary Medicine and Animal Science, Swedish University of Agricultural Sciences (SLU), Uppsala, Sweden.
*
All correspondence to: Ann-Sofi Bergqvist, Division of Comparative Reproduction, Obstetrics and Udder Health, Department of Clinical Sciences, Faculty of Veterinary Medicine and Animal Science, Swedish University of Agricultural Sciences (SLU), P.O. Box 7054, SE-750 07 Uppsala, Sweden. Tel: +46 18 671154. Fax: +46 18 673545. e-mail: [email protected]
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Summary

Glycosaminoglycans (GAGs) are present in the oviduct in which the major part of sperm capacitation occurs. In this study we have tested how capacitation of frozen-thawed bull spermatozoa is effected by exposure to different GAGs detectable or possibly present in oviductal fluid; i.e. heparin, hyaluronan, heparan sulphate, dermatan sulphate and chondroitin sulphate. Following exposure of different duration, the spermatozoa were stained with either Chlortetracycline (CTC) or merocyanine-540 and evaluated with epifluorescent light microscopy or flow cytometry, respectively. Heparin elicited a significant increase in the number of alive, capacitated spermatozoa, either expressed as higher merocyanine-540 fluorescence (p < 0.0001) or as B-pattern (p = 0.0021) in the CTC assay, during 4 h of incubation. When comparing the different GAG treatments one by one to the negative control in the flow cytometric study, only heparin and dermatan sulphate were significant (p < 0.0001) higher than the control at 0–30 min of incubation. Duration of incubation did not affect the proportion of capacitated spermatozoa when measured as merocyanine-540 fluorescence or CTC B-pattern, but the length of the incubation did affect the number of dead (Yo-PRO 1 positive) spermatozoa (p < 0.0001). Exposure to zona pellucida proteins significantly increased the proportion of acrosome reacted spermatozoa (p = 0.016). Both heparin and dermatan sulphate induce capacitation of frozen-thawed bull spermatozoa in vitro.

Keywords

CTCFlow cytometryGAGsSperm capacitation
Type
Research Article
Information
Zygote , Volume 15 , Issue 3 , August 2007 , pp. 225 - 232
Copyright
Copyright © Cambridge University Press 2007

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