Porcine Follicular Fluid Proteins Analysis
This study isolated porcine oocytes to determine the morphology and to classify porcine follicular fluid protein patterns by Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The ovaries of reproductive female pigs were obtained from local slaughterhouses in Nakorn Pathom Province, Thailand. The porcine oocytes complexes (pCOCs) and follicles were aspirated and collected by sterile technique. The pCOCs were collected from different follicles sizes. We found that the oocytes can be categorized into intact cumulus cells layer oocyte (Type 1), multi cumulus cells layer oocyte (Type 2), partial cumulus cells layer oocyte (Type 3), completely denuded oocyte (Type 4)and degenerated oocyte (Type 5). A total of 1,150 follicles were divided into 2 groups. Group 1 consists of small follicles with diameters of 1-3 mm (n = 701) were grouped into 5 types as described above, including 237, 118, 192, 124 and 30 oocytes, respectively. Group 2 was medium follicles with diameters of 4-6 mm (n = 449) contain 5 types were 174, 82, 90, 92 and 11 oocytes, respectively. This study also classified porcine follicular fluid protein patterns by SDS-PAGE. The results showed protein band sizes composed of > 180, 170, 160, 85, 70-65 and 32 kDa (kilodaltons). Moreover the samples were subjected to LC/MS/MS and the selected the band of protein at 65 kDa were serum albumin. These data are basic information which is useful for developing applications in biotechnology such as role in stimulating cell proliferation, fertilization or used the porcine secretion proteins for supplement in IVM/IVF technology.
Keywords - Porcine Oocytes, Protein Analysis, Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-Page).