Alterations of sperm DNA integrity during cryopreservation procedure and in vitro incubation of bull semen |
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| Authors: | KE Waterhouse A Gjeldnes A Tverdal PM De Angelis W Farstad M Håård E Kommisrud |
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| Institution: | 1. Team Semin, P.O. Box 8146 Dep., N-0033 Oslo, Norway;2. The Norwegian School of Veterinary Science, Department of Production Animal Clinical Science, P.O. Box 8146 Dep., N-0033 Oslo, Norway;3. The Norwegian School of Veterinary Science, Department of Basic Sciences and Aquatic Medicine, P.O. Box 8146 Dep. N-0033 Oslo, Norway;4. Institute of Pathology, Rikshospitalet-Radiumhospitalet HF, N-0027 Oslo, Norway;5. Svensk Avel, Örnsro, 532 94 Skara, Sweden;6. Geno Breeding and AI Association, N-2326 Hamar, Norway;1. Laboratory of Semen Biotechnology and Andrology, Center of Biotechnology in Animal Reproduction, Animal Reproduction Department, College of Veterinary Medicine and Animal Science (FMVZ), University of São Paulo (USP), Pirassununga, São Paulo, Brazil;2. Animal Reproduction Department, College of Veterinary Medicine and Animal Science (FMVZ), University of São Paulo (USP), São Paulo, Brazil;3. Laboratory of Theriogenology Dr. O. J. Ginther, Department of Veterinary Medicine, FZEA, University of São Paulo (USP), Pirassununga, São Paulo, Brazil;1. Department of Veterinary Medical Sciences (DIMEVET), University of Bologna, Bologna, Italy;2. AUB INFA National Institute of Artificial Insemination, Bologna, Italy;1. Department of Animal Production, College of Food and Agriculture Sciences, King Saud University, P.O. Box 2460, Riyadh 11451, Saudi Arabia;3. Department of Physiology, Faculty of Veterinary Medicine, Zagazig University, 44519 Zagazig, Egypt;4. Department of Biological Sciences, Faculty of Science, University of Jeddah, Saudi Arabia;5. Department of Biochemistry, Faculty of Veterinary Medicine, Zagazig University, Sharkia 44519, Egypt;1. Department of Zoology, Pir Mehr Ali Shah Arid Agriculture University, Rawalpindi 46300, Pakistan;2. Department of Wildlife Management, Pir Mehr Ali Shah Arid Agriculture University, Rawalpindi 46300, Pakistan;3. Buffalo Research Institute, Pattoki, District Kasur, 55050, Pakistan;4. Semen Production Unit, Qadirabad, District Sahiwal, Pakistan;5. Instituto de Biología y Medicinal Experimental (IBYME), National Research Council of Argentina (CONICET), Vuelta de Obligado 2490 (C1428ADN), Buenos Aires 1428ADN, Argentina;1. Department of Animal Reproduction, School of Veterinary Medicine, UNIRP – Centro Universitário de Rio Preto, Unidade Universitária II (Hospital Veterinário), Rodovia Br 153 (Transbrasiliana), Km 69, 15093-450, São José do Rio Preto, SP, Brazil;2. Department of Preventive Veterinary Medicine and Animal Reproduction, School of Agrarian Sciences and Veterinary Medicine, Univers Estadual Paulista, Jaboticabal, SP 14884-900, Brazil;3. School of Veterinary Medicine and Animal Science, University of São Paulo, Pirassununga, SP 13630-000, Brazil;4. School of Animal Science, Michigan State University, East Lansing, MI 48824, USA;5. School of Veterinary Medicine, Federal University of Uberlândia, Uberlândia, MG 38400-902, Brazil |
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| Abstract: | An association between sperm DNA integrity and fertility was recently shown for frozen–thawed Norwegian Red (NRF) bull semen diluted in skimmed milk egg yolk (SMEY). In general the fertility of NRF cattle is high, however, in comparison with NRF semen in SMEY, NRF semen diluted in Tris EY based extenders has shown reduced fertility. The aim of the present study was to do a split-sample comparison of sperm DNA integrity of NRF bull semen (n = 20) in SMEY and Triladyl® (Tris EY based) during routine cryopreservation procedure and during in vitro incubation of frozen–thawed semen in modified synthetic oviduct fluid (mSOF). In contrast to the high fertility of NRF cattle, Holstein cattle are experiencing a marked decline in fertility. Therefore, the present study also aimed to compare sperm DNA integrity of NRF (n = 20) and Holstein (n = 20) semen diluted in Triladyl® during in vitro incubation. The sperm DNA integrity was measured by susceptibility to in situ acid induced denaturation by the Sperm chromatin structure assay (SCSA). Compared to initial values of frozen neat semen, an increase in DNA damage was observed after dilution and cooling (5 °C) and after freezing–thawing of NRF semen in SMEY, but only after freezing–thawing for NRF semen diluted in Triladyl®. Sperm DNA damage of NRF semen increased during in vitro incubation in mSOF; the increase in percentage of spermatozoa with DNA damage was more prominent in SMEY than in Triladyl®, while the degree of damage was higher in Triladyl®, throughout the incubation period. However, while the correlation between DNA damage and sperm survival was negative in SMEY throughout the incubation period, a positive correlation was observed in Triladyl® after 9 h of incubation, indicating a higher presence of DNA damage in the live sperm population. In comparison with Holstein spermatozoa, the sperm DNA integrity of NRF semen reflected a better ability to withstand alterations induced during in vitro incubation in mSOF. In conclusion, sperm DNA integrity of NRF bull semen was altered during the cryopreservation procedure and in vitro incubation in mSOF. Dilution in Triladyl® maintained bull sperm DNA integrity better than dilution in SMEY. Furthermore, alterations in Holstein sperm DNA integrity was more pronounced during in vitro incubation in mSOF compared to NRF bull spermatozoa. |
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