Laparoscopy vs. laparotomy for embryo transfer to produce transgenic goats (Capra hircus)

다국어 초록 (Multilingual Abstract)

This study was performed to produce transgenic Korean
native goat (Capra hircus) by laparoscopic embryo transfer
(ET) to overcome the limitations of ET performed by
laparotomy. Transgenic embryos were produced by DNA
pronuclear microinjection of in vivo zygotes. The recipient
goats were synchronized for estrus by using an introvaginal
progesterone devices as a controlled internal drugreleasing
insert (CIDR) for 13 days and injection of 400 IU
PMSG 48 h before removal of the insert. Embryos were
transferred on day 3 and 4 after removal of the insert.
Recipient goats were deprived of feed for 48 h, then
suspended in a laparotomy cradle at an angle of 45o. After
obtaining a sufficient pneumoperitoneum, the laparoscope
and forceps were inserted abdominally through 5 mm
trocar sleeves. Examination of the ovaries and uterus was
performed and then 213 embryos were transferred into the
oviducts via the infundibula of 76 recipient goats. To
compare pregnancy rates, ET was also performed by
laparotomy in 82 recipient goats. The pregnancies in the
recipient goats were diagnosed by ultrasound on day 30
after embryo transfer. The pregnancy rate with
laparoscopic ET was significantly higher than with ET
performed by laparotomy (46.1% vs. 28.6%, p ＜ 0.05). In
addition, the pregnancy rates were compared between
ovulated and non-ovulated ovaries of the recipient goats in
the laparoscopic ET group. No significant difference was
observed between the pregnancy rates of ovulated and
non-ovulated ovaries (41.3% vs. 33.3%, p ＜ 0.05) suggesting
that ET may also be possible in non-ovulated recipients
through artificial rupture of Graafian follicles. These
results suggest that laparoscopic ET is a highly efficient
method for the transfer of goat embryos.

다국어 초록 (Multilingual Abstract)

This study was performed to produce transgenic Korean
native goat (Capra hircus) by laparoscopic embryo transfer
(ET) to overcome the limitations of ET performed by
laparotomy. Transgenic embryos were produced by DNA
pronuclear microinjection of in vivo zygotes. The recipient
goats were synchronized for estrus by using an introvaginal
progesterone devices as a controlled internal drugreleasing
insert (CIDR) for 13 days and injection of 400 IU
PMSG 48 h before removal of the insert. Embryos were
transferred on day 3 and 4 after removal of the insert.
Recipient goats were deprived of feed for 48 h, then
suspended in a laparotomy cradle at an angle of 45o. After
obtaining a sufficient pneumoperitoneum, the laparoscope
and forceps were inserted abdominally through 5 mm
trocar sleeves. Examination of the ovaries and uterus was
performed and then 213 embryos were transferred into the
oviducts via the infundibula of 76 recipient goats. To
compare pregnancy rates, ET was also performed by
laparotomy in 82 recipient goats. The pregnancies in the
recipient goats were diagnosed by ultrasound on day 30
after embryo transfer. The pregnancy rate with
laparoscopic ET was significantly higher than with ET
performed by laparotomy (46.1% vs. 28.6%, p ＜ 0.05). In
addition, the pregnancy rates were compared between
ovulated and non-ovulated ovaries of the recipient goats in
the laparoscopic ET group. No significant difference was
observed between the pregnancy rates of ovulated and
non-ovulated ovaries (41.3% vs. 33.3%, p ＜ 0.05) suggesting
that ET may also be possible in non-ovulated recipients
through artificial rupture of Graafian follicles. These
results suggest that laparoscopic ET is a highly efficient
method for the transfer of goat embryos.

참고문헌 (Reference)

1 Edmunds T, "Transgenically produced human antithrombin: structural and functional comparison to human plasma-derived antithrombin" 91 : 4561-4571, 1998

2 Ebert KM, "Transgenic production of a variant of human tissue-type plasminogen activator in goat milk: generation of transgenic goats and analysis of expression" 9 : 835-838, 1991

3 Wang B, "Transgenic goats produced by DNA pronuclear microinjection of in vitro derived zygotes" 63 : 437-443, 2002

4 Ebert KM, "Transgenic farm animals: progress report" 39 : 121-135, 1993

5 Denman J, "Transgenic expression of a variant of human tissue-type plasminogen activator in goat milk: purification and characterization of the recombinant enzyme" 9 : 839-843, 1991

6 Mutiga ER, "Transfer of sheep embryos through a laparoscope" 114 : 401-402, 1984

7 Clark AJ, "The mammary gland as a bioreactor: expression, processing, and production of recombinant proteins" 3 : 337-350, 1998

8 Kiessling AA, "Superovulation and embryo transfer in the dairy goat" 188 : 829-832, 1986

9 Kühholzer B, "Prokofiev MI, Ernst LK, Brem G. Laparoscopic recovery of pronuclearstage goat embryos" 142 : 40-42, 1998

10 Baldassarre H, "Production of transgenic goats by pronuclear microinjection of in vitro produced zygotes derived from oocytes recovered by laparoscopy" 59 : 831-839, 2003

11 Colman A, "Production of proteins in the milk of transgenic livestock: problems, solutions, and successes" 63 : 639S-645S, 1996

12 Houdebine LM, "Production of pharmaceutical proteins from transgenic animals" 34 : 269-287, 1994

13 Ko JH, "Production of biologically active human granulocyte colony stimulating factor in the milk of transgenic goat" 9 : 215-222, 2000

14 Tittel A, "New adhesion formation after laparoscopic and conventional adhesiolysis: a comparative study in the rabbit" 15 : 44-46, 2001

15 Fayrer-Hosken RA, "Laparoscopic oviductal transfer of in vitro matured and in vitro fertilized bovine oocytes" 32 : 413-420, 1989

16 Ebert KM, "Induction of human tissue plasminogen activator in the mammary gland of transgenic goats" 12 : 699-702, 1994

17 Takahashi Y, "In vitro development of bovine one-cell embryos: Influence of glucose, lactate, pyruvate, amino acids and vitamins" 37 : 963-978, 1992

18 Gootwine E, "Factors affecting success of embryo collection and transfer in a transgenic goat program" 48 : 485-499, 1997

19 Besenfelder U, "Endoscopic embryo collection and embryo transfer into the oviduct and the uterus of pigs" 47 : 1051-1060, 1997

20 Lee CS, "Embryo recovery and transfer for the production of transgenic goats from Korean native strain, Capra hircus aegagrus" 37 : 57-63, 2000

21 Koeman J, "Developmental competence of prepubertal and adult goat oocytes cultured in semi-defined media following laparoscopic recovery" 60 : 879-889, 2003

22 Rudolph NS, "Biopharmaceutical production in transgenic livestock" 17 : 367-374, 1999

23 Baldassarre H, "Advances in the production and propagation of transgenic goats using laparoscopic ovum pick-up and in vitro embryo production technologies" 57 : 275-284, 2002

24 McKelvey WA, "A simplified technique for the transfer of ovine embryos by laparoscopy" 117 : 492-494, 1985