Glutathione supplementation to semen extender improves the quality of frozen-thawed canine spermatozoa for transcervical insemination.
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.
Department of Animal Production Science, United Graduate School of Agricultural Science, Tokyo University of Agriculture and Technology; University Farm, Faculty of Agriculture, Utsunomiya University, Tochigi 321-4415, Japan.
Kazuko
K
Ogata
Aiko
A
Sasaki
Yuka
Y
Kato
Arisa
A
Takeda
Mikio
M
Wakabayashi
Borjigin
B
Sarentonglaga
Mio
M
Yamaguchi
Asuka
A
Hara
Rika
R
Fukumori
Yoshikazu
Y
Nagao
D000818:Animals; D015925:Cryopreservation / Q000379:methods / Q000662:veterinary; D004285:Dogs; D005978:Glutathione / Q000008:administration & dosage*; D007315:Insemination, Artificial / Q000379:methods / Q000662:veterinary*; D015227:Lipid Peroxidation / Q000187:drug effects; D008297:Male; D055101:Semen Analysis; D012662:Semen Preservation / Q000379:methods / Q000662:veterinary*; D013081:Sperm Motility / Q000187:drug effects; D013094:Spermatozoa / Q000187:drug effects*
D005978:Glutathione