Transgenic goats produced by DNA pronuclear microinjection of in vitro derived zygotes.
This study was undertaken to investigate various factors affecting the outcomes of in vitro fertilization (IVF) of oocytes retrieved by laparoscopic ovum pick-up (LOPU) technique from prepubertal and adult goats, as well as to evaluate the developmental competence of in vitro produced embryos. Oocyte-cumulus complexes recovered by LOPU from donors stimulated with gonadotrophins were matured in vitro. Fresh semen was used for IVF following various capacitation treatments. In vitro produced zygotes were either cultured to assess in vitro development or were transferred into recipients for full term development. The results indicated that successful IVF of the goat oocytes was affected by factors such as sperm capacitation treatment, oocyte quality, and abundance of cumulus cells on zona pellucida. Oocytes from both prepubertal and adult goats demonstrated similar full term developmental competence despite the fact that in vitro developmental rates were lower for prepubertal goats. The births of transgenic offspring demonstrated that the established LOPU-IVF technology combined with pronuclear microinjection can be successfully used to produce transgenic goats.
Wang
Bin
B
Nexia Biotechnologies Inc, Vaudreuil Dorion, Quebec, Canada. bwang@nexiabiotech.com
Baldassarre
H
H
Tao
T
T
Gauthier
M
M
Neveu
N
N
Zhou
J F
JF
Leduc
M
M
Duguay
F
F
Bilodeau
A S
AS
Lazaris
A
A
Keefer
C
C
Karatzas
C N
CN
D000818:Animals; D030801:Animals, Genetically Modified; D002467:Cell Nucleus / Q000235:genetics / Q000502:physiology*; D004247:DNA / Q000008:administration & dosage / Q000494:pharmacology*; D004624:Embryo Transfer; D005314:Embryonic and Fetal Development; D005307:Fertilization in Vitro / Q000379:methods*; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D006041:Goats / Q000196:embryology* / Q000235:genetics*; D015759:Ionomycin / Q000494:pharmacology; D008845:Microinjections; D009865:Oocytes / Q000187:drug effects / Q000378:metabolism*; D015053:Zygote / Q000166:cytology / Q000502:physiology*
D015759:Ionomycin; D004247:DNA