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Effect of GnRH injection timing in the production of pronuclear-stage zygotes used for DNA microinjection.
Effect of GnRH injection timing in the production of pronuclear-stage zygotes used for DNA microinjection.
- Source :
-
Zygote (Cambridge, England) [Zygote] 2004 Aug; Vol. 12 (3), pp. 257-61. - Publication Year :
- 2004
-
Abstract
- This study was aimed at developing a hormonal treatment protocol in order to optimize the proportion of pronuclear-stage embryos to be used for DNA microinjection in a goat transgenic founder production programme. A total of 46 adult BELE and 47 adult standard goats (1-5 years old) were used as donors and recipients, respectively. They were heat-synchronized using intravaginal sponges containing 60 mg medroxyprogesterone acetate for 10 days with an injection of 125 microg cloprostenol on the morning of the eighth day. Recipients were injected with 400 IU eCG at the time of sponge removal while donors received a total of 133 mg NIH-FSH-P1 (Folltropin-V) given twice daily in decreasing doses over 3 days starting 48 h before sponge removal. Ovulation was induced in donors by injecting 100 microg of GnRH at 24 h (GnRH24) or 36 h (GnRH36) after sponge removal. Embryo recovery was performed by oviduct flushing following a standard mid-ventral laparotomy procedure. The proportion of embryos in the pronuclear stage of development was higher in the GnRH36 group (90% vs 34%, p < 0.01). Embryos were microinjected with a DNA expression cassette followed by transfer to the oviduct of synchronized recipients. A higher, yet not statistically significant, pregnancy rate was found in the recipients transferred with pronuclear-stage embryos compared with those transferred with 2-cell-stage embryos (64% vs 37%, chi-square p = 0.06). One transgenic female founder was produced from the group of recipients transferred with pronuclear-stage microinjected embryos.
- Subjects :
- Animals
DNA administration & dosage
DNA metabolism
Female
Fertility Agents, Female administration & dosage
Goats
Gonadotropin-Releasing Hormone administration & dosage
Pregnancy
Pregnancy Outcome
Time Factors
Fertility Agents, Female pharmacology
Gonadotropin-Releasing Hormone pharmacology
Microinjections
Zygote drug effects
Subjects
Details
- Language :
- English
- ISSN :
- 0967-1994
- Volume :
- 12
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- Zygote (Cambridge, England)
- Publication Type :
- Academic Journal
- Accession number :
- 15521716
- Full Text :
- https://doi.org/10.1017/s0967199404002849