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Amphibian oocytes and embryos are classical models to study cellular and developmental processes. For these studies, it is often advantageous to visualize protein organization. However, the large size and yolk distribution make imaging of deep structures in amphibian zygotes challenging. Here we describe in detail immunofluorescence (IF) protocols for imaging microtubule assemblies in early amphibian development. We developed these protocols to elucidate how the cell division machinery adapts to drastic changes in embryonic cell sizes. We describe how to image mitotic spindles, microtubule asters, chromosomes, and nuclei in whole-mount embryos, even when they are hundreds of micrometers removed from the embryo's surface. Though the described methods were optimized for microtubule assemblies, they have also proven useful for the visualization of other proteins.
BAKER,
Remarks on the discovery of cell-division.
1951, Pubmed
BAKER,
Remarks on the discovery of cell-division.
1951,
Pubmed Becker,
Visualization of the cytoskeleton in Xenopus oocytes and eggs by confocal immunofluorescence microscopy.
2006,
Pubmed
,
Xenbase Coleman,
The Xenopus Cdc6 protein is essential for the initiation of a single round of DNA replication in cell-free extracts.
1996,
Pubmed
,
Xenbase Dent,
A whole-mount immunocytochemical analysis of the expression of the intermediate filament protein vimentin in Xenopus.
1989,
Pubmed
,
Xenbase Field,
Spindle-to-cortex communication in cleaving, polyspermic Xenopus eggs.
2015,
Pubmed
,
Xenbase Field,
Xenopus extract approaches to studying microtubule organization and signaling in cytokinesis.
2017,
Pubmed
,
Xenbase Gard,
Confocal microscopy and 3-D reconstruction of the cytoskeleton of Xenopus oocytes.
1999,
Pubmed
,
Xenbase Groen,
Purification and Fluorescent Labeling of Tubulin from Xenopus laevis Egg Extracts.
2016,
Pubmed
,
Xenbase Gurdon,
The use of Xenopus oocytes for the expression of cloned genes.
1983,
Pubmed
,
Xenbase KARASAKI,
Studies on amphibian yolk 1. The ultrastructure of the yolk platelet.
1963,
Pubmed KARASAKI,
STUDIES ON AMPHIBIAN YOLK. 5. ELECTRON MICROSCOPIC OBSERVATIONS ON THE UTILIZATION OF YOLK PLATELETS DURING EMBRYOGENESIS.
1963,
Pubmed Kieserman,
High-magnification in vivo imaging of Xenopus embryos for cell and developmental biology.
2010,
Pubmed
,
Xenbase Levy,
Nuclear size is regulated by importin α and Ntf2 in Xenopus.
2010,
Pubmed
,
Xenbase Lohka,
Formation in vitro of sperm pronuclei and mitotic chromosomes induced by amphibian ooplasmic components.
1983,
Pubmed
,
Xenbase Lohka,
Induction of nuclear envelope breakdown, chromosome condensation, and spindle formation in cell-free extracts.
1985,
Pubmed
,
Xenbase Lombard-Banek,
High-Sensitivity Mass Spectrometry for Probing Gene Translation in Single Embryonic Cells in the Early Frog (Xenopus) Embryo.
2016,
Pubmed
,
Xenbase Mayor,
Induction of the prospective neural crest of Xenopus.
1995,
Pubmed
,
Xenbase Mitchison,
Growth, interaction, and positioning of microtubule asters in extremely large vertebrate embryo cells.
2012,
Pubmed
,
Xenbase Murray,
Cell cycle extracts.
1991,
Pubmed Nguyen,
Spatial organization of cytokinesis signaling reconstituted in a cell-free system.
2014,
Pubmed
,
Xenbase Paddock,
Laser scanning confocal microscopy: history, applications, and related optical sectioning techniques.
2014,
Pubmed Peshkin,
On the Relationship of Protein and mRNA Dynamics in Vertebrate Embryonic Development.
2015,
Pubmed
,
Xenbase Reinsch,
Movement of nuclei along microtubules in Xenopus egg extracts.
1997,
Pubmed
,
Xenbase Sawin,
Mitotic spindle assembly by two different pathways in vitro.
1991,
Pubmed
,
Xenbase Sive,
Dejellying Xenopus laevis Embryos.
2007,
Pubmed
,
Xenbase Smits,
Global absolute quantification reveals tight regulation of protein expression in single Xenopus eggs.
2014,
Pubmed
,
Xenbase Ulrich,
[Confocal laser scanning microscopy].
2015,
Pubmed Wallingford,
Xenopus.
2010,
Pubmed
,
Xenbase Wühr,
Evidence for an upper limit to mitotic spindle length.
2008,
Pubmed
,
Xenbase Wühr,
A model for cleavage plane determination in early amphibian and fish embryos.
2010,
Pubmed
,
Xenbase Wühr,
Deep proteomics of the Xenopus laevis egg using an mRNA-derived reference database.
2014,
Pubmed
,
Xenbase Wühr,
The Nuclear Proteome of a Vertebrate.
2015,
Pubmed
,
Xenbase Wühr,
How does a millimeter-sized cell find its center?
2009,
Pubmed
,
Xenbase