An investigation of capacitation and the acrosome reaction in dog spermatozoa using a dual fluorescent staining technique.
The processes of capacitation and acrosomal exocytosis of dog spermatozoa in vitro have yet to be fully investigated. Firstly, we investigated the effectiveness of a technique for staining dog spermatozoa with the fluorescent labels Hoechst 33258 and chlortetracycline. A modified fluorescence microscopy staining method was shown to be effective for the assessment of both viability and functional status in this species. Secondly, the presence of the ionophore A23187 in culture medium was shown to promote capacitation and the acrosome reaction of dog spermatozoa. We have therefore established that this dual fluorescent staining method can be used for monitoring these events in the dog, and it may be useful in future studies of optimal in vitro culture conditions.
Department of Farm Animal and Equine Medicine and Surgery, Royal Veterinary College, University of London, Hatfield, UK. dhewitt@rvc.ac.uk
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Hewitt
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England
D000177:Acrosome / Q000502:physiology*; D000814:Aniline Compounds / Q000737:chemistry; D000818:Animals; D006690:Bisbenzimidazole / Q000737:chemistry; D000001:Calcimycin / Q000737:chemistry; D002751:Chlortetracycline / Q000737:chemistry; D004285:Dogs / Q000502:physiology*; D005456:Fluorescent Dyes / Q000737:chemistry*; D007476:Ionophores / Q000737:chemistry; D008297:Male; D008856:Microscopy, Fluorescence / Q000662:veterinary; D013075:Sperm Capacitation / Q000502:physiology*; D013094:Spermatozoa / Q000502:physiology*
D000814:Aniline Compounds; D005456:Fluorescent Dyes; D007476:Ionophores; D000001:Calcimycin; C002712:nigrosin; D006690:Bisbenzimidazole; D002751:Chlortetracycline