Long-term cryopreservation of sperm from Mandarin fish Siniperca chuatsi.
In order to develop cryopreservation techniques for long-term preserving the sperm of Mandarin fish Siniperca chuatsi, we examined the effects of various extender and cryopreservation on post-thaw motility. We found the optimal freezing procedures for the Mandarin fish sperm is diluting the semen in D-15 extender, chilling it to 4 degrees C, adding ME2SO to a final concentration of 10% (v/v), then transferring the semen in cryotubes, holding the cryotubes for 10 min at 6 cm (about -180 degrees C) above the surface of liquid nitrogen, for 5 min on the surface of liquid nitrogen, and finally plunged into liquid nitrogen. After thawed at 37 degrees C for 60s, the sperm had the highest post-thaw motility (96.00+/-1.73%). The optimal fertilization procedures for the frozen sperm is mixing the eggs with sperm, then adding 1 ml of swimming medium (SM=45 mM NaCl+5 mM KCl+20mM Tris-HCl, pH 8.0) immediately. At the sperm/egg ratio of 100,000:1, the fertilization rate and the hatching rate of the frozen sperm cryopreserved for 1 week or 1 year in liquid nitrogen (66.01+/-5.14% and 54.76+/-4.40% & 62.97+/-14.28% and 52.58+/-11.17%) were similar to that of fresh sperm (69.42+/-8.11% and 59.82+/-5.27%) (p>0.05). This is the first report that the Mandarin fish (S. chuatsi) sperm can successfully fertilized eggs after long-term cryopreservation.
Ding
Shuyan
S
Freshwater Fisheries Research Institute of Jiangsu Province, Nanjing 210017, PR China.
Ge
Jiachun
J
Hao
Chen
C
Zhang
Mingsheng
M
Yan
Weihui
W
Xu
Zhiqiang
Z
Pan
Jianlin
J
Chen
Songlin
S
Tian
Yongsheng
Y
Huang
Yahong
Y
D000818:Animals; D015925:Cryopreservation / Q000379:methods* / Q000662:veterinary; D003451:Cryoprotective Agents / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005260:Female; D005298:Fertility / Q000187:drug effects / Q000502:physiology; D005306:Fertilization / Q000187:drug effects / Q000502:physiology; D008297:Male; D010473:Perciformes / Q000502:physiology; D012662:Semen Preservation / Q000379:methods* / Q000662:veterinary; D013081:Sperm Motility / Q000187:drug effects; D013094:Spermatozoa; D013997:Time Factors
D003451:Cryoprotective Agents