title: Fertilisation of cryopreserved sperm and unfertilised quail ovum by intracytoplasmic sperm injection.
abstract: Intracytoplasmic sperm injection (ICSI) is an important technique in animal biotechnology for animal cloning and conservation of genetic resources, but has been a challenge for avian species. In the present study, we investigated the ability of cryopreserved quail spermatozoa to achieve fertilisation and embryo development. Female quail were killed 70-120min after previous oviposition to collect unfertilised oocytes from the oviduct. Fresh or cryopreserved-thawed spermatozoa were injected into the cytoplasm of unfertilised oocytes, and the manipulated oocytes were incubated in quail surrogate eggshells. Injection of fresh spermatozoa supplemented with inositol 1,4,5-trisphosphate (IP3) resulted in a significantly increased rate of embryo development compared with injection of fresh spermatozoa alone (90% vs 13%, respectively). Although >80% of embryos stopped cell division and development before Hamburger and Hamilton (HH) Stage 3, approximately 15% of embryos from the fresh sperm injection developed to past HH Stage 4, and one embryo survived up to HH Stage 39 (11 days of incubation). In the case of cryopreserved spermatozoa, the embryo development rate was 30% after ICSI, and this increased significantly to 74% with IP3supplementation. In conclusion, cryopreserved spermatozoa combined with ICSI followed by surrogate eggshell culture can develop quail embryos.
authors.affiliation: Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, and Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul 151-921, Korea.
authors.forename: Kyung Soo
authors.initials: KS
authors.lastname: Kang
authors.affiliation: Graduate School of International Agricultural Technology and Institute of Green-Bio Science and Technology, Seoul National University, Pyeongchang-gun, Gangwon-do 232-916, Korea.
authors.forename: Tae Sub
authors.initials: TS
authors.lastname: Park
authors.affiliation: Department of Animal Science and Technology, Chung-Ang University, Anseong 456-756, Korea.
authors.forename: Deivendran
authors.initials: D
authors.lastname: Rengaraj
authors.affiliation: Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, and Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul 151-921, Korea.
authors.forename: Hyung Chul
authors.initials: HC
authors.lastname: Lee
authors.affiliation: Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, and Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul 151-921, Korea.
authors.forename: Hong Jo
authors.initials: HJ
authors.lastname: Lee
authors.affiliation: Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, and Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul 151-921, Korea.
authors.forename: Hee Jung
authors.initials: HJ
authors.lastname: Choi
authors.affiliation: Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Shizuoka 422-8529, Japan.
authors.forename: Shusei
authors.initials: S
authors.lastname: Mizushima
authors.affiliation: Division of Animal Science, Faculty of Agriculture, Shinshu University, 8304 Minamiminowa, Kamiina, Nagano 399-4598, Japan.
authors.forename: Tamao
authors.initials: T
authors.lastname: Ono
authors.affiliation: Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, and Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul 151-921, Korea.
authors.forename: Jae Yong
authors.initials: JY
authors.lastname: Han
mesh_terms: D000818:Animals; D015925:Cryopreservation; D005260:Female; D005306:Fertilization; D008297:Male; D009865:Oocytes; D011784:Quail; D020554:Sperm Injections, Intracytoplasmic; D013094:Spermatozoa / Q000166:cytology*