Studying the roles of Aurora-C kinase during meiosis in mouse oocytes

Methods Mol Biol. 2013:957:189-202. doi: 10.1007/978-1-62703-191-2_13.

Abstract

We previously isolated Aurora-C (Aurkc/Aie1) in a screen for kinases expressed in mouse sperm and eggs. Aurora-C kinase was reported to be a chromosomal passenger protein that plays critical roles in chromosome alignment, segregation, kinetochore-microtubule attachment, and cytokinesis in female mouse meiosis. This chapter describes experimental approaches for examining the subcellular localization and function of Aurora-C kinase during female mouse meiosis, presenting detailed methods for introducing exogenous Aurora-C wild-type and kinase-dead mutant mRNAs into mouse oocytes by cytosolic microinjection, and preparing whole-mount meiotic oocytes and chromosome spreads for confocal immunofluorescence microscopy.

MeSH terms

  • Animals
  • Aurora Kinase C
  • Aurora Kinases
  • Cell Separation
  • Chromosomes, Mammalian / metabolism
  • Cytosol / enzymology
  • Enzyme Assays / methods*
  • Female
  • Fluorescent Antibody Technique
  • Meiosis*
  • Mice
  • Mice, Inbred C57BL
  • Microinjections
  • Oocytes / cytology*
  • Oocytes / enzymology*
  • Oocytes / metabolism
  • Ovary / cytology
  • Protein Serine-Threonine Kinases / genetics
  • Protein Serine-Threonine Kinases / metabolism*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Staining and Labeling
  • Transcription, Genetic

Substances

  • RNA, Messenger
  • Aurkc protein, mouse
  • Aurora Kinase C
  • Aurora Kinases
  • Protein Serine-Threonine Kinases