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Laser Capture Microdissection of Highly Pure Trabecular Meshwork from Mouse Eyes for Gene Expression Analysis

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A high-efficiency trichome collection system by laser capture microdissection.

Wei Qin¹, Yongpeng Li¹, Bowen Peng¹

¹Frontiers Science Center for Transformative Molecules, Joint International Research Laboratory of Metabolic and Developmental Sciences, Key Laboratory of Urban Agriculture (South) Ministry of Agriculture, Plant Biotechnology Research Center, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China.

Frontiers in Plant Science

|September 8, 2022

Summary

This summary is machine-generated.

Researchers developed an efficient laser capture microdissection (LCM) method to isolate glandular secretory trichomes (GSTs) from unfixed plant leaves. This technique enables the analysis of intact secondary metabolites and gene expression in specific plant cell populations.

Keywords:

Artemisia annua L.artemisinin glandular secretory trichome laser capture microdissection secondary metabolites

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Area of Science:

Plant Biology
Molecular Biology
Biochemistry

Background:

Trichomes are epidermal structures in plants, with glandular secretory trichomes (GSTs) producing valuable specialized metabolites.
Isolating trichomes is crucial for understanding gene function and discovering new compounds, but it is challenging and labor-intensive.

Purpose of the Study:

To develop an efficient method for isolating GSTs from unfixed plant leaves using laser capture microdissection (LCM).
To enable the analysis of secondary metabolites and gene expression within isolated GSTs.

Main Methods:

Utilized laser capture microdissection (LCM) to isolate glandular secretory trichomes (GSTs) from unfixed *Artemisia annua* leaves.
Employed Ultra-Performance Liquid Chromatography (UPLC) for metabolite analysis and quantitative Reverse Transcription Polymerase Chain Reaction (qRT-PCR) for gene expression analysis.

Main Results:

Successfully isolated 150 GSTs efficiently from *Artemisia annua* leaves.
Detected specific accumulation of secondary metabolites from a small number of isolated GSTs.
Confirmed high expression of GST-specific genes involved in artemisinin biosynthesis within the isolated GSTs.

Conclusions:

Developed an efficient LCM-based method for collecting pure GSTs from unfixed leaves, preserving metabolite integrity.
This method facilitates metabolomics research on specific plant cell populations and the discovery of novel secondary metabolites.