Zhang, C., Gong, F. C., Lambert, G. M., & Galbraith, D. W. (2005). Cell type-specific characterization of nuclear DNA contents within complex tissues and organs. Plant Methods, 1(1).
Abstract:
Background: Eukaryotic organisms are defined by the presence of a nucleus, which encloses the chromosomal DNA, and is characterized by its DNA content (C-value). Complex eukaryotic organisms contain organs and tissues that comprise interspersions of different cell types, within which polysomaty, endoreduplication, and cell cycle arrest is frequently observed. Little is known about the distribution of C-values across different cell types within these organs and tissues.Results: We have developed, and describe here, a method to precisely define the C-value status within any specific cell type within complex organs and tissues of plants. We illustrate the application of this method to Arabidopsis thaliana, specifically focusing on the different cell types found within the root.Conclusion: The method accurately and conveniently charts C-value within specific cell types, and provides novel insight into developmental processes. The method is, in principle, applicable to any transformable organism, including mammals, within which cell type specificity of regulation of endoreduplication, of polysomaty, and of cell cycle arrest is suspected. © 2005 Zhang et al; licensee BioMed Central Ltd.
Galbraith, D., Zhang, C., Barthelson, R. A., Lambert, G. M., & Galbraith, D. W. (2008). Global characterization of cell-specific gene expression through fluorescence-activated sorting of nuclei. Plant physiology, 147(1).
We describe a simple and highly effective means for global identification of genes that are expressed within specific cell types within complex tissues. It involves transgenic expression of nuclear-targeted green fluorescent protein in a cell-type-specific manner. The fluorescent nuclei are then purified from homogenates by fluorescence-activated sorting, and the RNAs employed as targets for microarray hybridization. We demonstrate the validity of the approach through the identification of 12 genes that are selectively expressed in phloem.
Galbraith, D., Harkins, K. R., Jefferson, R. A., Kavanagh, T. A., Bevan, M. W., & Galbraith, D. W. (1990). Expression of photosynthesis-related gene fusions is restricted by cell type in transgenic plants and in transfected protoplasts. Proceedings of the National Academy of Sciences of the United States of America, 87(2).
We have analyzed the expression of chimeric genes in populations of protoplasts isolated from the photosynthetic and nonphotosynthetic tissues within leaves of transgenic tobacco plants and separated by fluorescence-activated cell sorting. Expression of transcriptional gene fusions controlled by promoters from photosynthesis-associated genes showed a striking dependence on cell type. These patterns of expression were preserved when the gene fusions were transfected into normal (nontransgenic) tobacco leaf protoplasts.
Galbraith, D. W., Bourque, D. P., & Bohnert, H. J. (1995). Preface. Methods in Cell Biology, 50(C), xxi-xxii.
BIO5 Collaborators
David W Galbraith, Laurence Hurley
Galbraith, D. W. (1990). Chapter 48 Flow Cytometric Analysis of Plant Genomes. Methods in Cell Biology, 33(C), 549-562.
