Isolation of Xenopus oocytes

KAREN M. NEWMAN; TRISTÁN H. AGÜERO; MARY LOU KING

Early Development of Xenopus laevis: A Laboratory Manual

Cold Spring Harbor

Año: 2018; p. 10 - 18

Xenopusoocytes and oocyte extracts are the starting material for a variety ofexperimental approaches.  In Xenopus,oogenesis is divided into six stages with each stage identified by a Romannumeral (Dumont, 1972).  Early stages ofoocytes have been ideal to image RNA localization (Heinrich and Deshler, 2009;Gagnon and Mowry, 2010); and extracts from late stages of oocyte have been usedto study the cell cycle (Novak and Tyson, 1993; Iwabuchi et al., 2000) andsignal transduction (Crane and Ruderman, 2006). Oocytes are obtained bysurgical removal of the ovary from anesthetized females. Although oocytes maybe used while they remain within their ovarian follicle, it is more practicalto work with defolliculated oocytes. Defolliculation can be done either manually or enzymatically.  Here we present a protocol for the isolationand separation of oocytes at various developmental stages, and guidelines formaintaining oocytes in culture.