Mehaisen Gamal Mohamed Kamel, Viudes-de-Castro María Pilar, Vicente José Salvador, Lavara Raquel
Laboratorio de Biotecnología de la Reproducción, Departamento de Ciencia Animal, Universidad Politécnica de Valencia, Camino de Vera 14, 46071 Valencia, Spain.
Theriogenology. 2006 Apr 15;65(7):1279-91. doi: 10.1016/j.theriogenology.2005.08.007. Epub 2005 Sep 19.
This study aimed to evaluate the in vitro and in vivo viability of vitrified and non-vitrified embryos derived from eCG and FSH treatments in rabbit does. Ninety-six nulliparous does were randomly subjected to consecutive superovulation treatments with eCG (20 IU/kg body weight intramuscularly (i.m.), eCG group), FSH (3 x 0.6 mg/doe at 24 h intervals i.m., FSH group), or without superovulation treatment (control group). Does were artificially inseminated 3 days later and ovulation was induced immediately by hCG (75 IU/doe intravenous). Seven experimental groups were differentiated: first FSH and eCG treatment, second FSH and eCG treatment, eCG-interchanged group (does with previous FSH treatment), FSH-interchanged group (does with previous eCG treatments) and control group. Embryos were collected in vivo by laparoscopy 76-80 h post-insemination in the first and second recovery cycles and post mortem in the third recovery cycles. The ovulation rate was significantly higher in does treated with the first-FSH than in those treated with eCG or in control does (25.2+/-2.0 versus 19.2+/-1.4 to 11.0+/-1.5, and 12.2+/-1.2, first-FSH, first-eCG to second-eCG and control groups, respectively, P < 0.05). Significant differences were observed in the total recovery influenced by ovulation rate in each group (20.3+/-2.2 to 9.4+/-1.2, first-FSH to control groups). Embryo donor rate (donor with at least one normal embryo) was similar among groups with an overall of 75.1%. The number of normal embryos recovered per doe with at least one normal embryo increased significantly in relation to ovulation rate (17.7+/-2.2 to 8.41+/-3, first-FSH and control groups). The vitrification of embryos negatively affected their in vitro development to hatched blastocyst in all groups (88.1% versus 48%, P > 0.05). However, after embryo transfer, this negative effect was only observed in superovulated vitrified embryos (16.8 and 12.8% versus 39.4% total born rate from eCG, FSH and control vitrified groups, P < 0.05). Results indicated that the primary treatments with eCG or FSH increased the number of normal embryos recovered per donor doe, but these embryos are more sensitive to vitrification protocols.
本研究旨在评估经促性腺激素释放激素(eCG)和促卵泡素(FSH)处理的兔未受精卵母细胞来源的玻璃化和非玻璃化胚胎在体外和体内的活力。96只未生育的母兔被随机分为连续超数排卵处理组,分别为eCG组(肌肉注射20 IU/kg体重的eCG)、FSH组(每只母兔每隔24小时肌肉注射3×0.6 mg FSH)或未进行超数排卵处理的对照组。3天后对母兔进行人工授精,并立即通过静脉注射人绒毛膜促性腺激素(hCG,75 IU/只)诱导排卵。区分出7个实验组:首次FSH和eCG处理组、第二次FSH和eCG处理组、eCG交换组(先前接受FSH处理的母兔)、FSH交换组(先前接受eCG处理的母兔)和对照组。在第一个和第二个回收周期中,在授精后76 - 80小时通过腹腔镜收集体内胚胎,在第三个回收周期中通过剖检收集。首次接受FSH处理的母兔排卵率显著高于接受eCG处理的母兔或对照组母兔(分别为25.2±2.0、19.2±1.4、11.0±1.5以及12.2±1.2,首次FSH、首次eCG、第二次eCG和对照组,P < 0.05)。在每组中,受排卵率影响的总回收率存在显著差异(20.3±2.2至9.4±1.2,首次FSH组至对照组)。胚胎供体率(至少有一个正常胚胎的供体)在各组中相似,总体为75.1%。每只至少有一个正常胚胎的母兔回收的正常胚胎数量与排卵率显著相关(首次FSH组和对照组分别为17.7±2.2和8.41±3)。胚胎玻璃化对所有组中胚胎体外发育至孵化囊胚均有负面影响(88.1%对48%,P > 0.05)。然而,胚胎移植后,这种负面影响仅在超数排卵的玻璃化胚胎中观察到(eCG、FSH和对照组玻璃化胚胎的总出生率分别为16.8%、12.8%对39.4%,P < 0.05)。结果表明,用eCG或FSH进行的初次处理增加了每个供体母兔回收的正常胚胎数量,但这些胚胎对玻璃化方案更敏感。