12 Confocal Immunofluorescence Microscopy of Microtubules, Microtubule-Associated Proteins, and Microtubule-Organizing Centers during Amphibian Oogenesis and Early Development

Abstract

Publisher Summary During the past 5 years, the picture of the microtubule (MT) cytoskeleton of amphibian oocytes has changed dramatically, due in large part to refinements in the techniques for preserving and visualizing the oocyte cytoskeleton. Individual MTs are first identified in the cortex of Xenopus oocytes in samples prepared by rapid freezing. Subsequently, whole-mount immunocytochemistry and confocal immunofluorescence microscopy has revealed an extensive network of MTs extending throughout the cytoplasm of oocytes from both X. laevis and Rana pipiens . In addition, these techniques have revealed that dramatic changes in MT assembly, organization, and dynamics accompany distinct stages in oocyte differentiation, maturation, and early embryonic development. This chapter presents a view of the assembly and organization of the MT cytoskeleton during oogenesis and early development in the African frog, X. laevis , which is based on results obtained in laboratories around the world. Results obtained from other amphibian species are discussed when informative. In addition, some of the mechanisms that might serve to regulate MT organization and dynamics during amphibian oogenesis and early development are addressed, focusing on the potential contributions of microtubule organizing centers (MTOCs) and microtubule-associated proteins (MAPS).