title: Quantitative Microinjection of Morpholino Antisense Oligonucleotides into Mouse Oocytes to Examine Gene Function in Meiosis-I.
abstract: Specific protein depletion is a powerful approach for assessing individual gene function in cellular processes, and has been extensively employed in recent years in mammalian oocyte meiosis-I. Conditional knockout mice and RNA interference (RNAi) methods such as siRNA or dsRNA microinjection are among several approaches to have been applied in this system over the past decade. RNAi by microinjection of Morpholino antisense Oligonucleotides (MO), in particular, has proven highly popular and tractable in many studies, since MOs have high specificity of interaction, low cell toxicity, and are more stable than other microinjected RNAi molecules. Here, we describe a method of MO microinjection into the mouse germinal vesicle-stage (GV) oocyte followed by a simple immunofluorescence approach for examination of gene function in meiosis-I.
authors.affiliation: Centre de Recherche du Centre Hospitalier de l'Université de Montréal, 900 Rue St. Denis, Montreal, QC, Canada, H2X 0A9.
authors.forename: Shoma
authors.initials: S
authors.lastname: Nakagawa
authors.affiliation: Centre de Recherche du Centre Hospitalier de l'Université de Montréal, 900 Rue St. Denis, Montreal, QC, Canada, H2X 0A9. greg.fitzharris@umontreal.ca.|Department of Obstetrics and Gynecology, Université de Montréal, 3175, Ch. Côte-Sainte-Catherine, Montréal, QC, Canada, H3T 1C5. greg.fitzharris@umontreal.ca.
authors.forename: Greg
authors.initials: G
authors.lastname: FitzHarris
mesh_terms: D000818:Animals; D005260:Female; D008540:Meiosis / Q000235:genetics*; D051379:Mice; D008845:Microinjections / Q000295:instrumentation / Q000379:methods; D008856:Microscopy, Fluorescence; D060172:Morpholinos / Q000008:administration & dosage*; D009865:Oocytes / Q000166:cytology / Q000378:metabolism*
chemical_list: D060172:Morpholinos
keywords: Immunofluorescence; Meiosis-I; Microinjection; Morpholino antisense oligonucleotides; Mouse oocyte