Glutathione supplementation to semen extender improves the quality of frozen-thawed canine spermatozoa for transcervical insemination |
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Authors: | Kazuko OGATA Aiko SASAKI Yuka KATO Arisa TAKEDA Mikio WAKABAYASHI Borjigin SARENTONGLAGA Mio YAMAGUCHI Asuka HARA Rika FUKUMORI Yoshikazu NAGAO |
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Institution: | 1)Department of Animal Production Science, United Graduate School of Agricultural Science, Tokyo University of Agriculture and Technology, Tokyo 183-8509, Japan;2)University Farm, Faculty of Agriculture, Utsunomiya University, Tochigi 321-4415, Japan;3)East Japan Guide Dog Association, Tochigi 321-0342, Japan;4)Collaboration Center for Research and Development of Utsunomiya University, Tochigi 321-0904, Japan |
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Abstract: | The present study was conducted to evaluate whether supplementation of semen extender with glutathione (GSH) can maintain the quality of frozen-thawed canine spermatozoa. Eighteen ejaculates were obtained from 5 dogs and placed in extender (20% egg yolk, Tris, citric acid, lactose, raffinose, antibiotics and 6.5% glycerol) containing 0 (control), 2.5, 5, 7.5 or 10 mM GSH. The samples were cooled to 4 C and then frozen in liquid nitrogen vapor. Motility parameters of the sperm were evaluated at 0, 1, 2, 3, 4, 12 and 24 h after thawing. Sperm motility was higher in the 5 mM GSH group than in the control or 2.5 and 10 mM GSH groups; this effect was observed at 1 to 24 h after thawing (P < 0.05). The 5 mM GSH group had a higher sperm viability index at 12 and 24 h after thawing compared with the other groups (P < 0.05). Acrosome integrity, evaluated at 4 h after thawing, was greater in two of the GSH-treated groups (5 and 10 mM) compared with the control. Lipid
peroxidation (LP) levels immediately after thawing were lower in the 5 and 10 mM GSH groups compared with the control, while those at 12 h after thawing did not differ significantly. Frozen-thawed semen in the 5 mM GSH group was used for transcervical insemination of 4 bitches, resulting in delivery of 5 puppies from 2 bitches. These results indicate that supplementation of semen extender with 5 mM GSH was effective in improving motility, longevity and acrosomal integrity and inhibiting LP levels in post-thaw canine spermatozoa, without any adverse impacts on full-term development after transcervical insemination. |
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Keywords: | Canine sperm Cryopreservation Glutathione Transcervical insemination |
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