Follicular Stimulation and Laparoscopic Ovum Pick-up (LOPU) in Repeatedly Superovulated Korean Black Goats
Laparoscopic ovum pick-up (LOPU) is a convenient method for collecting oocytes in small ruminants. LOPU has the advantage of being a less invasive means of oocyte collection, thereby allowing for a repeated usage of the oocyte donor animals. A total of 25 Korean black goats were used in the winter season (December to February) and LOPU was applied to the goats which had been treated for superovulation more than two times during the last twelve months. Estrus was synchronized with an intravaginal insert containing 0.3 g progesterone for 10 to 12 days. Ovaries were hyperstimulated with eCG 1,000 IU oneshot, FSH with eCG (50 mg / 1,000 IU; 70 mg / 500 IU; 70 mg / 1,000 IU) oneshot or FSH multiple-shot with eCG oneshot (20 mg × 6 / 300 IU) given intramuscularly 72 h prior to LOPU. For these groups, the number of follicles (mean ± SEM) observed which developed to larger than 2 mm in diameter were 1.6 ± 2.5, 4.3 ± 3.1, 5.5 ± 4.2, 6.6 ± 2.1 and 8.8 ± 7.8, respectively. Oocytes were aspirated by using OPU needles and a vacuum pump. The overall oocyte retrieval rates were 41.4%. Oocytes were matured in TCM-199 supplemented with 10% (w/v) bovine serum albumin + 10 μg/ml FSH + 1 μg/ml 17β-estradiol for 27 h at 39℃ in 5% CO2 in air. Oocytes were parthenogenetically activated by ionomycin combined with 6-diethylaminopurine (6- DMAP). Total oocyte maturation and cleavage rate were 67.3% and 78.8%, respectively. In summary, LOPU is a useful oocyte collection method in Korean black goats that can provide immature oocytes for transgenesis or nuclear transfer.