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, 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
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DOI：
10.1105/tpc.008102
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发表时间：
2003-03-01
期刊：
影响因子：
11.6
通讯作者：
Schnable, PS
中科院分区：
文献类型：
--
作者：
Nakazono, M;Qiu, F;Schnable, PS
激光捕获显微切割（LCM）允许从组织切片一步获得大的均质细胞群体。在哺乳动物中，LCM已被用于对特定细胞类型进行cDNA微阵列和蛋白质组学研究。然而，LCM尚未应用于植物细胞，最有可能的是因为植物细胞壁使得难以将靶细胞与周围细胞分离，并且因为在制备冷冻切片时，冰晶可以在细胞之间的空气空间中形成。通过固定组织，在冷冻前使用冷冻保护剂，并使用粘合剂涂覆的载玻片系统，可以从乙醇：乙酸固定的玉米胚芽鞘中捕获大量（> 10，000）表皮细胞和维管组织（维管束和束鞘细胞）。从这些细胞中提取的RNA用T7 RNA聚合酶扩增，并用于包含类似于8800个玉米cDNA的芯片杂交。其中约250种优先在表皮细胞或血管组织中表达。这些结果表明，LCM和微阵列的组合，使其成为可行的进行高分辨率的全球基因表达分析的植物。这种方法有可能提高我们对不同植物细胞类型特异性生物过程的理解。
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 similar to8800 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.