Comparison between intracytoplasmic sperm injection and in vitro fertilisation employing oocytes derived from prepubertal goats.
The objective of this study was to compare the embryo development of prepubertal goat oocytes after ICSI and IVF procedures. Three experiments were carried out to achieve this objective. (1) An analysis of the efficiency of ICSI with or without chemical stimulation (5 microM ionomycin for 5 min and 2 mM 6-DMAP for 4 h). In this experiment, Sham and parthenogenetic oocyte groups were used as controls. (2) According to the results from experiment 1, we investigated the nuclear stage of zygotes obtained with ICSI and IVF, and their further embryo development. (3) We compared two embryo culture media (G1.3/G2.3 and TCM199 with granulosa cells) on the embryo development of zygotes obtained from ICSI and IVF procedures. Experiment 1 demonstrated that prepubertal goat oocytes needed additional chemical stimulation, after conventional ICSI, to form zygotes with male and female pronuclei (2PN). Experiment 2 showed that significantly higher percentages of -zygotes were found in ICSI-oocytes than IVF-oocytes (40.0 and 25.1%, respectively; P < 0.005). The percentage of embryos obtained and developed beyond the 8-cell stage was significantly higher for ICSI than for IVF and parthenogenetic embryos (22.8, 10.3 and 3.8%, respectively; P < 0.05). Experiment 3 showed that G1.3/G2.3 medium improved the embryo development of ICSI- and IVF-oocytes compared to co-culture with granulosa cells in TCM medium. The highest percentage of embryo development beyond 8-16 cells was found in ICSI-oocytes cultured in G1.3/G2.3 medium. However, a reduced number of morulae were found in this study.
Jiménez-Macedo
Ana-Raquel
AR
Departament de Ciència Animal i dels Aliments, Facultat de Veterinària, Universitat Autònoma de Barcelona, Bellaterra, Barcelona 08193, Spain.
Izquierdo
Dolors
D
Anguita
Begoña
B
Paramio
Maria-Teresa
MT
D000818:Animals; D001755:Blastocyst; D018920:Coculture Techniques / Q000662:veterinary; D047108:Embryonic Development; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods / Q000662:veterinary*; D006041:Goats / Q000196:embryology*; D008297:Male; D009028:Morula; D009865:Oocytes / Q000254:growth & development / Q000502:physiology*; D010312:Parthenogenesis / Q000502:physiology; D012741:Sexual Maturation; D020554:Sperm Injections, Intracytoplasmic / Q000379:methods / Q000662:veterinary*