Metabolic profiling of laser microdissected vascular bundles of Arabidopsis thaliana

Plant Methods

Julia Kehr¹  Oliver Fiehn²  Rajsree Mungur¹  Martina Schad¹ 

[1] Max-Planck-Institute of Molecular Plant Physiology, Department Lothar Willmitzer, 14424 Potsdam, Germany;UC Davis, 4321 GBSF Building Health Sciences Drive, Davis (CA) 95616, USA

关键词: vascular bundle;    metabolite profiling;    tissue-specific analysis;    GC-TOF MS;    laser microdissection;   

Others  :  823981
DOI  :  10.1186/1746-4811-1-2

 received in 2005-05-26, accepted in 2005-08-18,  发布年份 2005

【 摘 要 】

Background

Laser microdissection is a useful tool for collecting tissue-specific samples or even single cells from animal and plant tissue sections. This technique has been successfully employed to study cell type-specific expression at the RNA, and more recently also at the protein level. However, metabolites were not amenable to analysis after laser microdissection, due to the procedures routinely applied for sample preparation. Using standard tissue fixation and embedding protocols to prepare histological sections, metabolites are either efficiently extracted by dehydrating solvents, or washed out by embedding agents.

Results

In this study, we used cryosectioning as an alternative method that preserves sufficient cellular structure while minimizing metabolite loss by excluding any solute exchange steps. Using this pre-treatment procedure, Arabidopsis thaliana stem sections were prepared for laser microdissection of vascular bundles. Collected samples were subsequently analyzed by gas chromatography-time of flight mass spectrometry (GC-TOF MS) to obtain metabolite profiles. From 100 collected vascular bundles (~5,000 cells), 68 metabolites could be identified. More than half of the identified metabolites could be shown to be enriched or depleted in vascular bundles as compared to the surrounding tissues.

Conclusion

This study uses the example of vascular bundles to demonstrate for the first time that it is possible to analyze a comprehensive set of metabolites from laser microdissected samples at a tissue-specific level, given that a suitable sample preparation procedure is used.

【 授权许可】

   
2005 Schad et al; licensee BioMed Central Ltd.

【 图 表 】

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