Abstract
The microtubule cytoskeleton plays important roles in a number of cellular processes including cell division, establishing and maintaining cell architecture and polarity, and intracellular trafficking. The identification and characterization of factors required for the proper functioning of the microtubule cytoskeleton have been aided by approaches that combine sensitive and rapid methods for high-resolution optical imaging, such as confocal microscopy, with the powerful genetics available in model organisms. Here we present methods for confocal imaging of live and fixed tissues of the nematode C. elegans, a model organism that has been employed with great success to study the microtubule cytoskeleton and its roles in cell division and cell polarity.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
(2004) Centrosomes in Development and Disease. Edited by Erich A. Nigg copyright 2004 Wiley-VCH Verlag GmbH &Co. KGaA, Weinheim ISBN 3-527-30980-2
Gard DL, Cha BJ, Schroeder MM (1995) Confocal immunofluorescence microscopy of microtubules, microtubule-associated proteins, and microtubule-organizing centers during amphibian oogenesis and early development. Curr Top Dev Biol 31:383–431
Bowerman B (2001) Cytokinesis in the C. elegans embryo: regulating contractile forces and a late role for the central spindle. Cell Struct Funct 26:603–607
Goldstein B (2000) Embryonic polarity: a role for microtubules. Curr Biol 10:R820–R822
Gonczy P (2002) Mechanisms of spindle positioning: focus on flies and worms. Trends Cell Biol 12:332–339
Crittenden SL, Kimble J (1999) Confocal methods for Caenorhabditis elegans. Methods Mol Biol 122:141–151
Miller DM, Shakes DC (1995) Immunofluorescence microscopy. Methods Cell Biol 48:365–394
Aroian RV, Field C, Pruliere G, Kenyon C, Alberts BM (1997) Isolation of actin-associated proteins from Caenorhabditis elegans oocytes and their localization in the early embryo. EMBO J 16:1541–1549
Seydoux G, Dunn MA (1997) Transcriptionally repressed germ cells lack a subpopulation of phosphorylated RNA polymerase II in early embryos of Caenorhabditis elegans and Drosophila melanogaster. Development 124:2191–2201
Schedl T (1997) Developmental genetics of the germ line. In: Riddle DL, Blumenthal T, Meyer BJ, Priess JR (eds) C. elegans II. Cold Spring Harbor Laboratory Press, Plainview, NY, pp 241–269
L'Hernault SW (1997) Spermatogenesis. In: Riddle DL, Blumenthal T, Meyer BJ, Priess JR (eds) C. ELEGANS II. Cold Spring Harbor Laboratory Press, Plainview, NY, pp 271–294
Albertson DG, Thomson JN (1993) Segregation of holocentric chromosomes at meiosis in the nematode, Caenorhabditis elegans. Chromosome Res 1:15–26
Verbrugghe KJ, White JG (2007) Cortical centralspindlin and G alpha have parallel roles in furrow initiation in early C. elegans embryos. J Cell Biol 120:1772–1778
O'Connell KF, Maxwell KN, White JG (2000) The spd-2 gene is required for polarization of the anteroposterior axis and formation of the sperm asters in the Caenorhabditis elegans zygote. Dev Biol 222:55–70
Golden A, Sadler PL, Wallenfang MR, Schumacher JM, Hamill DR, Bates G, Bowerman B, Seydoux G, Shakes DC (2000) Metaphase to anaphase (mat) transition-defective mutants in Caenorhabditis elegans. J Cell Biol 151:1469–1482
L'Hernault SW, Roberts TM (1995) Cell biology of nematode sperm. Methods Cell Biol 48:273–301
Acknowledgments
We thank Jill Schumacher for sharing her modification of the Seydoux and Dunn protocol, Mary Kosinski and Michael Miller for their modifications of the gonad suspension method, Penny Sadler for her sperm squash protocol, and Catherine Kemp, Murali Addepalli, and Anna Burrows for critically evaluating the manuscript.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media New York
About this protocol
Cite this protocol
O’Connell, K.F., Golden, A. (2014). Confocal Imaging of the Microtubule Cytoskeleton in C. elegans Embryos and Germ Cells. In: Paddock, S. (eds) Confocal Microscopy. Methods in Molecular Biology, vol 1075. Humana Press, New York, NY. https://doi.org/10.1007/978-1-60761-847-8_13
Download citation
DOI: https://doi.org/10.1007/978-1-60761-847-8_13
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-58829-351-0
Online ISBN: 978-1-60761-847-8
eBook Packages: Springer Protocols