Abstract
The use of antibodies to visualize the distribution and subcellular localization of gene products powerfully complements genetic and molecular analysis of gene function in Caenorhabditis elegans. Double and triple staining protocols are particularly useful for several reasons. First, colocalization of proteins either within tissues or at a subcellular level can be examined. Second, costaining with stage-specific or tissue-specific markers can define the timing and tissue specificity of antigen expression. For these types of studies it is useful to be able to collect data from multiple fluorescence wavelengths simultaneously. A confocal microscope equipped with a krypton/argon laser can simultaneously detect up to three different antigens. Using a confocal microscope it is also possible to collect a series of optical sections through a sample that allows observation of changes in distribution of the antigen in different focal planes of the tissue or cell.
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References
Strome, S. and Wood, W. B. (1982) Immunofluorescence visualization of germline-specific cytoplasmic granules in embryos, larvae, and adults of Caenorhabditis elegans. Proc. Natl. Acad. Sci. USA 79, 1558–1562.
Albertson, D. G. (1984) Formation of the first cleavage spindle in nematode embryos. Dev. Biol. 101, 61–72.
Bowerman, B., Draper, B. W., Mello, C. C., and Priess, J. F. (1993) The maternal gene skn-1 encodes a protein that is distributed unequally in early C. elegans embryos. Cell 74, 443–452.
Strome, S. (1986) Fluorescence visualization of the distribution of microfilaments in gonads and early embryos of the nematode Caenorhabditis elegans. J. Cell Biol. 103, 2241–2252.
Crittenden, S. L., Troemel, E. R., Evans, T. C., and Kimble, J. (1994) GLP-1 is localized to the mitotic region of the C. elegans germ line. Development 120, 2901–2911.
Harlow, E. and Lane, D. (1988) Antibodies: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.
Sulston, J. and Hodgkin, J. (1988) Methods, in The Nematode Caenorhabditis elegans (W. B. Wood, ed.), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, pp. 587–606.
Finney, M. and Ruvkun, G. (1990) The unc-86 gene product couples cell lineage and cell identity in C. elegans. Cell 63, 895–905.
Miller, D. M. and Shakes, D. C. (1995) Immunofluorescence microscopy, in Caenorhabditis elegans: Modern Biological Analysis of an Organism (H. F. Epstein and D. C. Shakes, eds.). Academic Press, San Diego, pp. 365–394.
Evans, T. C., Crittenden, S. L., Kodoyianni, V., and Kimble, J. (1994) Translational control of maternal glp-1 mRNA establishes an asymmetry in the C. elegans embryo. Cell 77, 183–194.
Lin, R., Thompson, S., and Priess, J. R. (1995) pop-1 encodes an HMG box protein required for the specification of a mesoderm precursor in early C. elegans embryos. Cell 83, 599–609.
Lin, R., Hill, R. J., and Priess J. R. (1998) POP-1 and anterior-posterior fate decisions in C. elegans embryos. Cell 92, 229–239.
Goldstein, B. and Hird, S. N. (1996) Specification of the anteroposterior axis in Caenorhabditis elegans. Development 122, 1467–1474.
Henderson, S. T., Gao, D., Lambie, E. J., and Kimble, J. (1994) lag-2 may encode a signaling ligand for the GLP-1 and LIN-12 receptors of C. elegans. Development 120, 2913–2924.
Francis, R., Barton, M. K., Kimble, J., and Schedl, T. (1995) gld-1, a tumor suppressor gene required for oocyte development in Caenorhabditis elegans. Genetics 139, 579–606.
Waddle, J. A., Cooper, J. A., and Waterston, R. H. (1994) Transient localized accumulation of actin in Caenorhabditis elegans blastomeres with oriented asymmetric divisions. Development 120, 2317–2328.
Guo, S. and Kemphues, K. J. (1995) par-1, a gene required for establishing polarity in C. elegans embryos, encodes a putative Ser/Thr kinase that is asymmetrically distributed. Cell 81, 611–620.
Fire, A., Xu, S., Montgomery, M., Kostas, S. A., Driver, S. E., and Mello, C. C. (1998) Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 391, 806–811.
Crittenden, S. L., Rudel, D., Binder, J., Evans, T. C., and Kimble, J. (1997) Genes required for GLP-1 asymmetry in the early Caenorhabditis elegans embryo. Dev. Biol. 181, 36–46.
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© 1999 Humana Press Inc., Totowa, NJ
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Crittenden, S.L., Kimble, J. (1999). Confocal Methods for Caenorhabditis elegans . In: Paddock, S.W. (eds) Confocal Microscopy Methods and Protocols. Methods in Molecular Biology™, vol 122. Humana Press. https://doi.org/10.1385/1-59259-722-X:141
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DOI: https://doi.org/10.1385/1-59259-722-X:141
Publisher Name: Humana Press
Print ISBN: 978-0-89603-526-3
Online ISBN: 978-1-59259-722-2
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