Laser-capture microdissection, a tool for the global analysis of gene expression in specific plant cell types: identification of genes expressed differentially in epidermal cells or vascular tissues of maize.
Laser-capture microdissection (LCM) allows for the one-step procurement of large homogeneous populations of cells from tissue sections. In mammals, LCM has been used to conduct cDNA microarray and proteomics studies on specific cell types. However, LCM has not been applied to plant cells, most likely because plant cell walls make it difficult to separate target cells from surrounding cells and because ice crystals can form in the air spaces between cells when preparing frozen sections. By fixing tissues, using a cryoprotectant before freezing, and using an adhesive-coated slide system, it was possible to capture large numbers (>10,000) of epidermal cells and vascular tissues (vascular bundles and bundle sheath cells) from ethanol:acetic acid-fixed coleoptiles of maize. RNA extracted from these cells was amplified with T7 RNA polymerase and used to hybridize a microarray containing approximately 8800 maize cDNAs. Approximately 250 of these were expressed preferentially in epidermal cells or vascular tissues. These results demonstrate that the combination of LCM and microarrays makes it feasible to conduct high-resolution global gene expression analyses of plants. This approach has the potential to enhance our understanding of diverse plant cell type-specific biological processes.
Nakazono
Mikio
M
Department of Agronomy, Iowa State University, Ames, Iowa 50011, USA.
Qiu
Fang
F
Borsuk
Lisa A
LA
Schnable
Patrick S
PS
D020346:Aquaporins / Q000235:genetics / Q000378:metabolism; D020869:Gene Expression Profiling / Q000295:instrumentation / Q000379:methods*; D018506:Gene Expression Regulation, Plant; D007834:Lasers; D037102:Lectins / Q000235:genetics / Q000378:metabolism; D008031:Lignin / Q000096:biosynthesis / Q000235:genetics; D008668:Metallothionein / Q000235:genetics / Q000378:metabolism; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation / Q000379:methods; D019441:Plant Epidermis / Q000235:genetics*; D018547:Plant Stems / Q000235:genetics*; D015203:Reproducibility of Results; D012765:Shikimic Acid / Q000378:metabolism; D003313:Zea mays / Q000235:genetics*
D020346:Aquaporins; D037102:Lectins; D012765:Shikimic Acid; D008031:Lignin; D008668:Metallothionein