Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
???displayArticle.abstract???
RNA localization in the Xenopus oocyte is responsible for the establishment of polarity during oogenesis as well as the specification of germ layers during embryogenesis. However, the inability to monitor mRNA localization in live vertebrate oocytes has posed a major barrier to understanding the mechanisms driving directional transport. Here we describe a method for imaging MS2 tagged RNA in live Xenopus oocytes to study the dynamics of RNA localization. We also focus on methods for implementing and analyzing FRAP data. This protocol is optimized for imaging of the RNAs in stage II oocytes but it can be adapted to study dynamics of other molecules during oogenesis. Using this approach, mobility can be measured in different regions of the oocyte, enabling the direct observation of molecular dynamics throughout the oocyte.
Axelrod,
Mobility measurement by analysis of fluorescence photobleaching recovery kinetics.
1976, Pubmed
Axelrod,
Mobility measurement by analysis of fluorescence photobleaching recovery kinetics.
1976,
Pubmed Bertrand,
Localization of ASH1 mRNA particles in living yeast.
1998,
Pubmed Chang,
Localization of RNAs to the mitochondrial cloud in Xenopus oocytes through entrapment and association with endoplasmic reticulum.
2004,
Pubmed
,
Xenbase Dumont,
Oogenesis in Xenopus laevis (Daudin). I. Stages of oocyte development in laboratory maintained animals.
1972,
Pubmed
,
Xenbase Gagnon,
Directional transport is mediated by a Dynein-dependent step in an RNA localization pathway.
2013,
Pubmed
,
Xenbase Gagnon,
Visualization of mRNA localization in Xenopus oocytes.
2011,
Pubmed
,
Xenbase Gagnon,
Molecular motors: directing traffic during RNA localization.
2011,
Pubmed
,
Xenbase Hocine,
Single-molecule analysis of gene expression using two-color RNA labeling in live yeast.
2013,
Pubmed Medioni,
Principles and roles of mRNA localization in animal development.
2012,
Pubmed Messitt,
Multiple kinesin motors coordinate cytoplasmic RNA transport on a subpopulation of microtubules in Xenopus oocytes.
2008,
Pubmed
,
Xenbase Mowry,
Vegetal messenger RNA localization directed by a 340-nt RNA sequence element in Xenopus oocytes.
1992,
Pubmed
,
Xenbase Rino,
Imaging dynamic interactions between spliceosomal proteins and pre-mRNA in living cells.
2014,
Pubmed Sprague,
Analysis of binding reactions by fluorescence recovery after photobleaching.
2004,
Pubmed
