Effectiveness Of Rabbit Anti-Alpha-Tubulin (Sc-69970, Santa Cruz Biotechnology) To Detect The Protein In Mouse Oocytes And Eggs

The 14-3-3 (YWHA) proteins are central mediators in important cellular events. We previously reported that all seven mammalian 14-3-3 isoforms are expressed in mouse oocytes and eggs and that, 14-3-3 eta (YWHAH) accumulates and co-localizes in the region of meiotic spindle in mouse eggs matured in vivo. Therefore, I investigated the distribution of 14-3-3 eta in mouse oocytes matured in vitro. In this project, I selected the rat anti-alpha-tubulin antibody to detect alpha-tubulin comprising microtubulues, as I had effectively used this antibody earlier to observe this protein in mouse eggs matured in vivo (De et al., 2012). In mouse eggs matured in vitro, 14-3-3 eta was observed to gradually accumulate in the regions of meiotic metaphase II spindles and co-localize with alpha-tubulin. This was demonstrated by indirect immunocytochemical staining of mouse eggs matured in vitro. These findings indicate that 14-3-3 eta may be required for normal meiotic spindle formation during maturation of mouse oocytes, in part by interacting with alpha-tubulin to regulate the assembly of microtubules.