Cytokeratin distribution and expression during the maturation of mouse germinal vesicle oocytes after vitrification

Abstract

Cytokeratin (CK) is a type of the cytoskeleton that increases cell stabilization during oocyte maturation. This study was conducted to investigate the effect of vitrification on the distribution and expression of CK during mouse oocyte maturation. Germinal vesicle (GV) oocytes were randomly allocated into three groups: (1) untreated (fresh), (2) exposed to vitrification solution (VS) without being plunged into liquid nitrogen (CPA exposure), or (3) vitrified using the open-pulled straw (OPS) method (vitrification). The oocytes were then incubated for 0h, 6h (metaphase I (MI) stage), and 10h (metaphase II (MII) stage). The CK distribution in the oocytes at the GV, MI, and MII stages was observed by immunofluorescence, and the expression at the MII stage oocytes derived from vitrified GV oocytes was detected by Western blotting. The CK distribution in the GV oocytes (88.5%) displayed a cortical pattern in the fresh group, whereas a granular pattern was mainly found at the MI (86.7%) and MII (93.5%) stages. In the CPA exposure group, 90.3% of the GV oocytes were observed to display the cortical pattern, and 69.2% of the MI oocytes and 92.9% of the MII oocytes showed the granular pattern. In the vitrification group, most oocytes (GV, 88.9%; MI, 100%; MII, 93.3%) exhibited the cortical pattern. The CK fluorescence intensities of the MII stage oocytes in both the fresh (59.27) and CPA exposure (60.05) group were significantly higher than that of the vitrification (26.53) group (p<0.05). Western blotting showed that the CK expression in the MII oocytes derived from vitrified GV oocytes was significantly lower (p<0.05) than in the control. In conclusion, OPS vitrification affects the normal CK distribution pattern during oocyte maturation and results in decreased CK expression in MII oocytes derived from vitrified GV oocytes.