ABSTRACT
BACKGROUND: Canine embryo cryopreservation and subsequent transfer are relevant in the use of reproductive technologies. OBJECTIVE: The purpose of this study is the identification and quantification of the gene expression BAX and Bcl2, AQP3, Na+/K+ ATPase alpha-1 and beta-1 and LIFr in canine embryos obtained in vivo and after freezing. MATERIALS AND METHODS: For the collection of embryos, the bitches were identified at pro-estrous until the detection of 80-90% superficial cells. After that, they were artificially inseminated with fresh semen. The embryos were collected after ovariohysterectomy. RNA was extracted and amplified, and embryos were randomly distributed into fresh (Fr) and frozen/thawed (Ft) groups. RESULTS: Eighteen blastocysts were collected from three bitches. Genes BAX, AQP3 and LIFr did not differ among the studied groups. CONCLUSION: We suggest, through these results, that the genes BAX, Bcl2, AQP3, Na + / K + ATPase alpha-1 and beta-1 and LIFr were expressed in canine blastocysts collected in vivo and after slow freezing cryopreservation.
Subject(s)
Blastocyst , Cryopreservation , Embryo, Mammalian , Gene Expression , Animals , Dogs , FreezingABSTRACT
The objectives were to evaluate the reexpansion blastocoele rate, post-thaw viability, and in vitro development of canine blastocysts cryopreserved by slow freezing in 1.0 m glycerol (GLY) or 1.5 m ethylene glycol (EG). Fifty-one in vivo-produced canine blastocysts were randomly allocated in two groups: GLY (n = 26) and EG (n = 25). After thawing, embryos from M0 were immediately stained with the fluorescent probes propidium iodide and Hoechst 33 342 to evaluate cellular viability. Frozen-thawed embryos from M3 and M6 were cultured in SOFaa medium + 10% FCS at 38.5°C under an atmosphere of 5% CO(2) with maximum humidity, for 3 and 6 days, respectively, and similarly stained. The blastocoele reexpansion rate (24 h after in vitro culture) did not differ between GLY (76.5%) and EG (68.8%). Post-thaw viable cells rate were not significantly different between GLY and EG (66.5 ± 4.8 and 57.3 ± 4.8, respectively, mean ± SEM), or among M0 (62.3 ± 5.7%), M3 (56.9 ± 6.0%), and M6 (66.5 ± 6.0%). In conclusion, canine blastocysts cryopreserved by slow freezing in 1.0 m glycerol or 1.5 m ethylene glycol, had satisfactory blastocoele reexpansion rates, similar post-thawing viability, and remained viable for up to 6 days of in vitro culture.
