Single cell RNA-Seq analysis provide deeper molecular insights into abnormal maturation of tubastatin A-treated mouse oocytes

In our previous study, we observed that Tub-A treatment induced an increased level of the acetylation of alpha-tubulin, and a failure of spindle migration and actin cap formation. Most Tub-A treated oocytes were arrested in an MI-like or a GVBD-like stage and decreased mTOR (spindle formation factor) and mDia1 (inhibitor of actin assembly) in an HDAC6 expression-dependent manner. We wonder if TubA only affects HDAC6 expression. Therefore the purpose of this study is to clarify the cause of the phenomenon through RNA-seq analysis. Overall design: Oocyte collection and culture: GV oocytes from Hybrid B6D2F1 (C57BL/6 x DBA) female mice (6-8 weeks old) were selected and maintained in M16 medium. Tubastatin A treatment: Tubastatin A HCl (Catalogue no. 27108) was obtained from BPS Bioscience (San Diego, USA). The compound was dissolved in DMSO and further diluted in saline to the 10 and 20uM concentration.