TY  - JOUR
TI  - Use of soluble sperm extract to improve cloning efficiency in zebrafish
AU  - Prukudom, Sukumal
AU  - Perez, GloriaI.
AU  - Cibelli, JoseB.
AU  - Siripattarapravat, Kannika
T2  - The International Journal of Developmental Biology
AB  - During somatic cell nuclear transfer (SCNT), egg activation is required to initiate embryonic development. In zebrafish cloning, the reconstructed egg is activated by exposing it to hypotonic water. Egg activation using water-only is not capable of activating the same intracellular calcium release as fertilization which is required for proper embryonic development. Here we test whether the use of soluble sperm extract (SSE) can properly modulate the activation of reconstructed eggs during SCNT. We microinjected SSE from genomic-inactivated zebrafish sperm into unfertilized eggs and reconstructed eggs right after somatic cell nuclear transfer. We also evaluated the most effective approach for SSE microinjection. Microinjection of SSE (with 0.68 mg/ml of protein concentration) into non-activated eggs through the micropyle induced parthenogenetic development beyond the blastula stage, whereas all water-only activated eggs failed to enter the cleavage period. Microinjection of SSE at 1 mg/ml of protein concentration into non-activated reconstructed egg improved the developmental rate of cloned embryos in comparison to non-injected control clones. The cumulative survival time of cloned embryos injected with SSE was significantly longer than reconstructed eggs activated following sham injection (P
PY  - 2019
DO  - 10.1387/ijdb.180367ks
VL  - 63
IS  - 6-7
SP  - 287
EP  - 293
J2  - Int. J. Dev. Biol.
LA  - en
SN  - 0214-6282
SN  - 1696-3547
UR  - https://ijdb.ehu.eus/article/180367ks
Y2  - 2024/11/23/12:27:18
ER  -