Abstract
The central nervous system (CNS) of the third instar larva is the most convenient tissue in which to examine both mitotic chromosomes and the diploid interphase nucleus. It contains a mixture of cell types at various degrees of differentiation. Here, methods for examining the diploid nuclei of the third instar larva are presented. One method employs BrdU incorporation to label nuclei during the S-phase. A second uses fluorescence in situ hybridization (FISH) to determine where certain sequences are located. Both can be used on either mitotic chromosomes or interphase nuclei. These two methods can be used by themselves or in combination. When used in combination, a researcher can identify which cells have recently undergone the S-phase and examine the specific positions of various chromosomal regions in the interphase nucleus.
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References
Marshall, W. F., Fung, J. C., and Sedat, J. W. (1997) Deconstructing the nucleus: global architecture from local interactions. Curr. Opin. Genet. Dev. 7, 259–263
Foe, V. E., Odell, G. M., and Edgar, B. A. (1993) Mitosis and morphogenesis in the Drosophila embryo: point and counterpoint, in The Development of Drosophila melanogaster (Bate, M. and Martinez Arias, A., eds.), Cold Spring Harbor Laboratory Press, Plainview, NY, Vol. 1, pp. 149–300.
Ashburner, M. (1989) Drosophila: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
Gatti, M., and Bonaccorsi, S., and Pimpinelli, S. (1994) Looking at Drosophila mitotic chromosomes. Methods Cell Biol. 44, 371–391.
Marshall, W. F., Dernburg, A. F., Harmon, B., Agard, D. A., and Sedat, J. W. (1996) Specific interactions of chromatin with the nuclear envelope: positional determination within the nucleus in Drosophila melanogaster. Mol. Biol. Cell 7, 825–842.
Csink, A. K. and Henikoff, S. (1998) Large-scale chromosomal movements during interphase progression in Drosophila. J. Cell Biol. 143, 13–22.
Agard, D. A., Hiraoka, Y., Shaw, P., and Sedat, J. W. (1989) Fluorescence microscopy in three dimensions. Methods Cell Biol. 30, 353–377.
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© 2004 Humana Press Inc., Totowa, NJ
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Csink, A.K. (2004). Analysis of Chromosomes of the Larval CNS by FISH and BrdU Labeling. In: Henderson, D.S. (eds) Drosophila Cytogenetics Protocols. Methods in Molecular Biology, vol 247. Humana Press. https://doi.org/10.1385/1-59259-665-7:343
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DOI: https://doi.org/10.1385/1-59259-665-7:343
Publisher Name: Humana Press
Print ISBN: 978-1-58829-050-2
Online ISBN: 978-1-59259-665-2
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