Identification of Protein Content of Immature Canine Oocytes.

The use of biotechnologies in wild canids, such as in vitro oocyte maturation (IVM) and fertilization (IVF), has received little attention. One of the main reasons is the difficulty in finding a relevant in vitro model, and domestic dogs (Canis lupus familiaris) may be a viable alternative. In vitro oocyte maturation in dogs is not well established, but canine oocytes can spontaneously restart meiosis through adaptations of bovine IVM protocols resulting in maturation rates of 10%-39%. To continue with the development of IVM and IVF protocols, the use of analytical techniques is necessary, but they require a minimum amount of protein. Oocyte protein content has been previously reported in different species. Unfortunately, the protein content in the canine oocyte remains unknown. In the present study, two different Bradford-based assays were used to determine the protein content in the immature oocyte of the domestic canines. To determine the protein content, 1500 immature oocytes were recovered from the ovaries of bitches and analyzed with Pierce 660 nm Protein Assay and Quant-iT Protein Assay Kit. Oocyte protein content average with Pierce 660 nm assay was 0.222 µg/oocyte, while for the Quant-iT Protein Assay Kit, the average was 0.205 µg/oocyte. Knowledge of the amount of protein contained in the canine immature oocyte will favour the design and development of experimental work with greater certainty and efficiency, as well as making efficient use of the samples, which will favour progress in the knowledge of the IVM of canine oocytes.