Assessment of nuclear membrane dynamics using anti-lamin staining offers a clear cut evidence of germinal vesicle breakdown in buffalo oocytes

Abstract

Importance of germinal vesicle breakdown (GVBD) in the development chronology of oocyte development can be gauged from the fact that it has been the target of several studies attempting to improve oocyte competence. Manipulation of this event rests on its precise and explicit documentation. Pertinently, we decided to compare three different methods for their efficacy and precision in assessing GVBD. Immature buffalo oocytes were subjected to GVBD inhibition using Roscovitine (Ros) and Cilostamide (Cil) under different concentrations of each as well as their combination, along with control. After 24 h of inhibition, chromatin assessment was done using aceto-orcein staining, hoechst staining and anti-lamin staining. Hoechst staining and anti-lamin staining proved to be easier and less time consuming when compared to aceto-orcein, which was cumbersome and lengthy. Aceto-orcein and hoechst staining were equally ambiguous, while antilamin staining was most precise, accurate and clear in characterizing an oocyte as germinal vesicle (GV) or GVBD. We conclude by stating that anti-lamin staining is an easy and specific technique for assessing GVBD in buffalo oocytes.