Superovulation Response and Quality of Embryos Recovered from Cattle after a Single Subcutaneous Injection of FSH Dissolved in Polyethylene Glycol

PEG 융해 FSH 투여방법에 따른 소 과배란 유기 및 수정란이식 효율

  • Published : 2002.04.01

Abstract

This study was carried out to assess the effect of superovulation response and quality of embryos recovered from donor cows after a single subcutaneous injection of FSH dissolved in polyethylene glycol (PEG). Cows were allocated into control and 3 experimental treatment groups. In control, cows were injected intramuscularly 50 mg FSH twice daily for 4 days. Group 1 were injected subcutaneously with a single dose of 400 mg FSH dissolved in 30% PEG solution. Group 2 were injected subcutaneously with a single dose of 200 mg FSH dissolved in 30% PEG solution. Finally in group 3, cows were injected twice 200 mg FSH dissolved in 30% PEG solution by subcutaneous. Superovulation was initiated by injection of FSH between Day 8 and 14 of the estrus cycle (Day 0, the day of estrus), and followed by injection of 25 mg PGF$_2$$\alpha$ at 48 h after first FSH injection. Cows were then artificially inseminated (AI) with semen twice at 48 and 60 h after PGF$_2$$\alpha$ injection. At 7 days after the second AI, embryos collected non-surgically by flushing the uterine horns and were counted and compared morphologically as being transferable and degenerated among different superovulation treatments. Furthermore, progesterone and estradiol -17 $\beta$ in plasma were measured by radioimmunoassay following different treatments at given days All cows of treated groups were observed heat. but control group was showed 77.8%. Superovulation response was observed as 77.8, 87.5, 88.9, and 100% in control, Groups 1, 2 and 3 The mean number of corpus lutea (CL) detected in Group 1 were 19.6, which was, respectively significantly (P<0.05) higher than those of other groups (11.1, 13.4 and 7.6, respectively). However, there did not differ on the mean number of total embryos recovered and of transferable embryos between control and treated groups.

본 실험에서는 한우에 있어서 PEG를 이용한 FSH의 투여 방법에 따른 과배란 유기 방법 및 수정란 이식의 효율을 조사하였다. 과배란 처리는 대조구를 포함하여 네 그룹으로 나누어 비교 실험하였으며, 대조구는 12시간 간격으로 1일 2회, 4일간 50 mg의 FSH를 근육주사, 처리구 1은 30% PEG에 용해시킨 400 mg의 FSH를 1회 피하주사, 처리구 2는 30% PEG에 용해시킨 200 mg의 FSH를 1회 피하주사, 그리고 처리구 3은 30% PEG에 용해 시킨 200 mg의 FSH를 2회 피하주사하였다. 발정일을 0일로 하여 8~14일째 FSH로 과배란 처리를 하였으며 인공수정은 PGF$_2$$\alpha$ 25 mg 투여 후 48시간과 60시간에 외부적인 발정에 상관없이 실시하였고, 2차 인공수정 후 7일째 비외과적인 방법을 이용하여 난자 또는 수정란을 회수하였으며 황체 수와 총 회수란 수를 조사하고 회수된 수정란을 등급 분류하여 이식 가능한 수정란 수를 조사하였다. 혈중 progesterone과 estradiol-17 $\beta$ 농도는 RIA를 이용하여 분석하였다. 본 실험에서 나타난 결과를 요약하면 다음과 같다. 1. 발정 발현은 대조구 (77.8%)를 제외한 모든 처리구에서 100% 나타났으며 과배란 유기는 대조구에서 77.8%, 처리 1구는 87.5%, 처리 2구는 88.9%, 처리 3구는 100% 이루어졌다. 2. 처리구 1에서 황체수가 평균 19.6개로 확인되었으며, 이는 대조구와 처리구 2, 3의 평균 황체수 11.1, 13.4, 9.6로 비교하였을 때 유의적으로(P<0.05) 높게 나타났다. 그러나 총회수란과 이식 가능한 수정란 수에서는 유의적인 차이는 없었다. 총 회수란 수는 대조구와 각 처리구에서 평균 9.1, 10.9, 9.4 그리고 6.9개였고, 이식 가능한 수정란 수는 굉균 5.1, 5.9, 7.3 그리고 4.6개였다. 3. 회수된 난자 또는 수정란의 등급을 분류하였을 때,'Excellent'로 분류되는 수정란의 비율은 처리구 3이 가장 높았으며 처리구 1이 가장 낮았다 (52.1%와 30.3%). 회수된 난자중 'Unfertilized'로 분류되는 난자는 처리구 2에서 가장 낮았다(4.0%).

Keywords

References

  1. Akbar AM, Nett TM and Niswendwr GD. 1974. Metabolic clearance and secretion rates of gonadotrophins at different stages of the oestrus cycle in ewes. Endocrinology, 94:1318-1324 https://doi.org/10.1210/endo-94-5-1318
  2. Bo GA, Hockley D, Tribulo H, Jofre F, Tribulo R, Busso N, Barth AD and Mapletoft RJ. 1991. The effect of dose schedule and route of administration on superovulatory response to Folltropin-V in the cow. Theriogenology, 35:186 https://doi.org/10.1016/0093-691X(91)90162-7
  3. Bo GA, Hockley DK, Nasser LF and Mapletoft RJ. 1994. Superovulatory response to a single subcutaneous injection of Folltropin-V in beef cattle. Theriogenology, 42:963-975 https://doi.org/10.1016/0093-691X(94)90119-4
  4. Callessen H, Greve T and Hyttel P. 1986.Preovulatory endocrinology and oocyte maturation in superovulated cattle. Theriogenology, 25:71-86 https://doi.org/10.1016/0093-691X(86)90184-6
  5. Dattena M, Vespignani S, Branca A, Gallus M, Ledda S, Naitana S and Cappai P. 1994. Superovulatory response and quality of embryos recovered from anestus ewes after a single injection of porcine FSH dissolved in polyvinylpyrrolidone. Theriogenology, 2:235-239
  6. Demoustier MM, Becker J-F, Van Der Zwalmen P, Clossect J, Gillard J-L and Ectors F. 1998. Determination of porcine plasma folltropin levels during superovulation treatment in cows. Theriogenology, 30:379-386
  7. Edward LM, Rahe CH, Griffin JL, Wolfe DF, Marple DN, Cummins KA and Pitchett JF. 1987. Effect of transportation stress on ovarian function in superovulated hereford heifers. Theriogenology, 28:291-299 https://doi.org/10.1016/0093-691X(87)90016-1
  8. Gonzalez A, Lussier JG, Caruthers TD, MurphyBD and Mapletoft RJ. 1990. Superovulation of beef heifer with Folltropin-V: a new FSH preparation containing reduced LH activity. Theriogenology, 33:519 https://doi.org/10.1016/0093-691X(90)90509-R
  9. Gwazdauskas FSW. 1972. Adrenocorticotropin alte ration of bovine peripheral plasma concentration of cortisol, corticosterone and progesterone. J. Dairy Sci., 55:1165-1169 https://doi.org/10.3168/jds.S0022-0302(72)85640-6
  10. Hansen PJ, Drost RM, Rivera FF, Paula-Lopes, AI-Katanani YM, Krininger CE and Chase CC. 2001. Adverse impact of heat stress on embryo production: causes and strategies for mitigation. Theriogenology, 55:91-103 https://doi.org/10.1016/S0093-691X(00)00448-9
  11. Lindsell CE, Rajkumar K, Manning AW, Emery SK, Mapletft RJ and Murphy BD. 1986. Variability in FSH-LH ratios among batches of commercially available gonadotrophins. Theriogenology, 25:167 https://doi.org/10.1016/0093-691X(86)90221-9
  12. Lindner GM and Wright RW Jr. 1983. Bovine embryo morphology and evaluation. Theriogenology, 20:407-416 https://doi.org/10.1016/0093-691X(83)90201-7
  13. Lopez-Sebastian A, Gomez-Brunet A, Lishman AW, Johnson SK. and Inskeep EK. 1993. Modification by propylene glycol of ovulation rate in ewes in response to a single injection of FSH. J. Reprod Fert., 99:437-442 https://doi.org/10.1530/jrf.0.0990437
  14. Monniaux D, Chupin D and Saummande J. 1983. Superovulatory responses of cattle. Theriogenology, 19:55-81 https://doi.org/10.1016/0093-691X(83)90124-3
  15. Murphy DB, Mapletoft R, Manns J and Humphrey WD. 1984. Variability in gonadotrophin preparations as a factor in superovulatory response. Theriogenoloy, 21:117-125 https://doi.org/10.1016/0093-691X(84)90311-X
  16. Nasser LF, Adams GP, Bo GA and Mapletoft RJ. 1993. Ovarian superstimulatory response relative to follicular wave emergence in heifers. Theriogenology, 40:713-724 https://doi.org/10.1016/0093-691X(93)90207-L
  17. Shea BF, Janzen RE and McDermand DF. 1984. Seasonal variation in reponse to stimulation and related embryo transfer procedures in Alberta over a nine year period. Theriogenology, 21:186-195 https://doi.org/10.1016/0093-691X(84)90316-9
  18. Stoebel DP and Moberg GP. 1982. Repeated acutestress during the follicular phase and luteinizing hormone surge of dairy heifers. J. Dairy Sci., 65:92-96 https://doi.org/10.3168/jds.S0022-0302(82)82157-7
  19. Suzuki T. 1993. Bovine embryo transfer and related techniques. Mol. Reprod. Dev., 36:236-237 https://doi.org/10.1002/mrd.1080360218
  20. Takedomi T, Aogagi Y, Konish M, Kishi H, Taya K, Watanabe G and Sasamoto S. 1993. Superovulation in Holstein heifers by a single injection of porcine FSH dissolved in polyviny-Ipyrrolidone. Theriogenology, 39:327 https://doi.org/10.1016/0093-691X(93)90182-5
  21. Takedomi T, Aogagi Y, Konish M, Kishi H, Taya K, Watanabe G and Sasamoto S. 1995. Superovulation in Holstein heifers by a single subcutaneous injection of porcine FSH dissolved in polyvinylpyrrolidone. Theriogenology, 43:1259-1268 https://doi.org/10.1016/0093-691X(95)00097-R
  22. Tibier M. 2000. The IETS statistics of embryo transfers in livestock in the world for the year 1999: A new record for bovine in vivo-derived embryos transferred. A report of the IETS data retrieval committee 24-28
  23. Tribulo H, Jofre F, Carcedo J, Alonso A, Tribulo R and Bo GA. 1993. Superovulation in Bosindicus cattle with a single subcutaneous injection of commercial pituitary extracts. Theriogenology, 39:331. abstr https://doi.org/10.1016/0093-691X(93)90186-9
  24. Walsh JH, Mantovani R, Duby RT, Overstrom EW, Dobrinsky JR, Enright WJ, Roche JF and Boland MP. 1993. The effects of once or twice daily injections of p-FSH on superovulatory response in heifers. Theriogenology, 40:313-321 https://doi.org/10.1016/0093-691X(93)90269-B
  25. Willmott N, Saunders J, Bo GA, Palasz A, Pierson RA and Mapletoft RJ. 1990. The effect of FSH/LH ratio in pituitary extracts on superovulatory response in the cow. Theriogenology, 33:347 https://doi.org/10.1016/0093-691X(90)90771-K
  26. Wolfenson D, Lew BJ, Thatcher WW, Graber Y and Meidan R. 1997. Seasonal and acute heat stress effects on steroid production by dominant follicles in cows. Anim. Reprod. Sci., 47:9-19 https://doi.org/10.1016/S0378-4320(96)01638-7
  27. Yamamoto M, Ooe M, Kawaguchi M and Suzuki T. 1994. Superovulation in the cow with a single intramuscular injection of FSH dissolved in polyvinylpyrrolidone. Theriogenology, 41:747-755 https://doi.org/10.1016/0093-691X(94)90184-K
  28. 손동수, 류일선, 박성재, 2001. 소 수정란이식(제 7판) 12-13
  29. 임석기, 우재석, 전기준, 장선식, 강수원, 윤상기, 손동수. 1998(a). 한우에 있어서 PEG에 용해시킨 Folltropin-V의 1회 피하주사에 의한 다배란유기. 한국수정란이식학회지, 13:207-212
  30. 임석기, 전기준, 우제석, 최재관, 양보석, 오성종, 윤상보. 1998(b). 한우에 있어서 다양한 용매에 용해시킨 FSH의 1회 주사에 의한 난소반응. 한국수정란이식학회지, 13(3):213-217
  31. 최상용, 노규진, 최창용, 강태영, 윤희준, 손우진, 이효종, 박충생. 1996. 토끼에서 FSH제의 Single Injection에 따른 과배란 유기 효과. 한국수정란이식학회지, 11:211-216