Fertilisation of cryopreserved sperm and unfertilised quail ovum by intracytoplasmic sperm injection
Kyung Soo Kang A , Tae Sub Park B , Deivendran Rengaraj C , Hyung Chul Lee A , Hong Jo Lee A , Hee Jung Choi A , Shusei Mizushima D , Tamao Ono E and Jae Yong Han A F G
+ Author Affiliations
- Author Affiliations
A 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.
B Graduate School of International Agricultural Technology and Institute of Green-Bio Science and Technology, Seoul National University, Pyeongchang-gun, Gangwon-do 232-916, Korea.
C Department of Animal Science and Technology, Chung-Ang University, Anseong 456-756, Korea.
D Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Shizuoka 422-8529, Japan.
E Division of Animal Science, Faculty of Agriculture, Shinshu University, 8304 Minamiminowa, Kamiina, Nagano 399-4598, Japan.
F Institute for Biomedical Sciences, Shinshu University, 8304 Minamiminowa, Kamiina, Nagano 399-4598, Japan.
G Corresponding author. Email: jaehan@snu.ac.kr
Reproduction, Fertility and Development 28(12) 1974-1981 https://doi.org/10.1071/RD15126
Submitted: 30 March 2015 Accepted: 2 June 2015 Published: 6 July 2015
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 70–120 min 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 IP3 supplementation. In conclusion, cryopreserved spermatozoa combined with ICSI followed by surrogate eggshell culture can develop quail embryos.
Additional keyword: surrogate eggshell.
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