An investigation of capacitation and the acrosome reaction in dog spermatozoa using a dual fluorescent staining technique |
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| Affiliation: | 1. Laboratório de Fungos Dimórficos Patogênicos, Departamento de Microbiologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, SP, Brazil;2. Laboratório de Quimioterapia Antifúngica, Departamento de Microbiologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, Av. Prof. Lineu Prestes 1374, 05508-900, ICB II, Lab 150, São Paulo, SP, Brazil;3. Departments of Medicine and Microbiology & Immunology, Albert Einstein College of Medicine, New York, NY, USA;1. Institute of Reproductive and Stem Cell Engineering, Basic Medicine College, Central South University, Changsha Hunan 410005, China;2. Clinical Research Center for Reproduction and Genetics in Hunan Province, Reproductive and Genetic Hospital of CITIC-Xiangya, ChangshaHunan 410005, China;3. Department of Parasitology, Xiangya School of Medicine, Central South University, ChangshaHunan 410005, China;4. Hunan Provincial Center for Disease Control and Prevention, Changsha Hunan 410005, China;5. Clinical Laboratory, Third Xiangya Hospital, Central South University, Changsha Hunan 410013, China;6. National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China;7. Center for Biosafety Mega-Science, Chinese Academy of Sciences, Wuhan 430071, China;8. Hunan New Outbreak Infectious Disease Prevention and Treatment Workstation of Chinese Academy of Medical Sciences Changsha, Hunan 410005, China |
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| Abstract: | 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. |
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