Production of transgenic goats by pronuclear microinjection of in vitro produced zygotes derived from oocytes recovered by laparoscopy

Baldassarre, Hernán; Wang, Bin; Kafidi, N.; Gauthier, Mélanie; Neveu, N.; Lapointe, J.; Sneek, L.; Leduc, Martin; Duguay, F; Zhou, Jiankang; Lazaris, Anthoula; Karatzas, C.N.

doi:10.1016/s0093-691x(02)01128-7

Cited by 59 publications

(51 citation statements)

References 19 publications

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“…The GnRH24 protocol may result in similar results of embryo development at flushing if embryo collection is advanced to 60 h after sponge removal, but this protocol may be more difficult to schedule. Improvements in the efficiency in the production of transgenic goats by pronuclear microinjection have recently been proposed by our group (Baldassarre et al, , 2003. This improvement is based on DNA microinjection of zygotes produced after in vitro maturation and fertilization of immature oocytes recovered by laparoscopy from gonadotropin-treated donor goats.…”

Section: Discussionmentioning

confidence: 99%

Effect of GnRH injection timing in the production of pronuclear-stage zygotes used for DNA microinjection

Baldassarre¹,

Wang²,

Gauthier³

et al. 2004

Zygote

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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 R 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 µg 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 µg 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.

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Section: Discussionmentioning

confidence: 99%

Effect of GnRH injection timing in the production of pronuclear-stage zygotes used for DNA microinjection

Baldassarre¹,

Wang²,

Gauthier³

et al. 2004

Zygote

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“…For these reasons, the use of dairy goats as a bioreactor animal is of interest. There have been several reports on the production of transgenic goats (Ebert et al 1991, Ebert and Schindler 1993, Gootwine et al 1997) and the feasibility of the large-scale production for the industrial application has been confirmed (Baldassarre et al 2003, Baldassarre and. The human Granulocyte Colony Stimulating Factor (hG-CSF) is a hematopoietic growth factor that stimulates the proliferation and the differentiation commitment of neutrophil precursor cells, and enhances some of the functional properties of mature neutrophils (Morstyn and Burgess 1988).…”

Section: Introductionmentioning

confidence: 99%

Production of transgenic goat (Capra hircus) with human Granulocyte Colony Stimulating Factor (hG-CSF) gene in Brazil

Freitas

Serova

Andreeva

et al. 2007

An. Acad. Bras. Ciênc.

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In order to produce transgenic goats with hG-CSF, a total of 24 adult Saanen and 48 adult undefined breed goats were used as donors and recipients, respectively. Donors were estrus-synchronized with vaginal sponges and superovulated by a treatment with 200 mg FSH given twice daily in decreasing doses over 3 days starting 48 h before sponge removal. Ovulation was induced by injecting 100 µg GnRH 36 h after sponge removal. The recipients also received an estrus synchronization treatment. Donors were mated with fertile Saanen bucks and, approximately 72 h after sponge removal, zygotes were recovered surgically by flushing oviducts. The recovered zygotes were briefly centrifuged to a reliable visualization of the pronuclei. The DNA construct containing hG-CSF gene flanked by goat and bovine αs1-casein sequences was injected into pronuclei of 129 zygotes. The microinjected embryos (3-6 per female) were transferred to 27 recipients. Ten recipients became pregnant and 12 kids were born. One transgenic male founder was identified in the group of kids. This is the first report of a birth of a transgenic goat in Latin America.

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“…This method is often called as laparoscopic ovum pick-up (LOPU) and seems to be very efficient for the recovery and quality of the COCs obtained in sheep and goats. Usually, in sheep, a short repeated gonadotrophin treatment is followed by LOPU [35][36][37][38]. In contrast, goats can be stimulated with a 'one shot' regime of gonadotrophins (one shot of follicular stimulating hormone (FSH) plus equine chorionic gonadotrophin (eCG), compared to the traditional multi-dose and F), insertion of trocar and cannula, cannula is left on the abdomen through which endoscope is inserted into the abdomen; (G), insertion of the endoscope through the cannula to visualize the ovaries, corpora lutea and reproductive tract; (H), a uterine horn is punctured close to the utero-tubular junction using a blunt-end 18 Gauge needle, embryos are loaded into an ET catheter and transferred into the lumen.…”

Section: Oocyte Collection and In Vitro Maturationmentioning

confidence: 99%

“…Further application of in vitro fertilization in obtaining sex pre-determined offspring will be an advantage for the livestock industry [47,20]. In vitro fertilization of prepubertal oocytes and oocytes recovered from LOPU [37] are also important areas to be developed further.…”

Section: In Vitro Fertilizationmentioning

confidence: 99%

“…Many transgenic animals have been produced for biomedical research interests as well as to test their application in livestock production [78]. There are many techniques available for the production of transgenic animals, such as pronuclear microinjection [79,80,37], sperm-mediated transgenesis [81] and somatic cell nuclear transfer [82,65,83]. During the last decade, research has focused on producing transgenic animals which secrete recombinant proteins in their milk.…”

Section: Production Of Transgenic Sheep and Goatsmentioning

confidence: 99%

See 1 more Smart Citation

Recent Advances in Reproductive Biotechnologies in Sheep and Goat

Alexander¹,

Mastromonaco²,

King³

2010

J Veterinar Sci Technolo

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Production of transgenic goats by pronuclear microinjection of in vitro produced zygotes derived from oocytes recovered by laparoscopy

Cited by 59 publications

References 19 publications

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

Production of transgenic goat (Capra hircus) with human Granulocyte Colony Stimulating Factor (hG-CSF) gene in Brazil

Recent Advances in Reproductive Biotechnologies in Sheep and Goat

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