Laser capture microdissection (LCM) for analysis of gene expression in specific tissues during embryonic epithelial-mesenchymal transformation

Ali Nawshad, D. LaGamba, B. R. Olsen, Elizabeth D. Hay

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

The analysis of gene expression in developing organs is a valuable tool for the assessment of genetic fingerprints during the various stages of tissue differentiation and epithelial-mesenchymal transformation (EMT). However, the variety of differentiating cells and the close association of epithelial and mesenchymal cells makes it difficult to extract protein and mRNA from specific cells and tissue and, thus, to assign expressed genes to specific cell populations. We report here the analysis of LEF1 mRNA in epithelial and mesenchymal cells isolated by LCM from different stages of EMT during development of the mouse palate and describe our techniques in detail. By applying a laser capture microdissection (LCM) technique and real-time polymerase chain reaction, we were able to determine mRNA levels that accurately reflect changes in gene expression in specific cells. The sensitivity of the technique is remarkable. Indeed, the mRNAs can be detected for many proteins too low in abundance to stain with antibodies. These techniques will enable embryologists to collect homogeneous groups of cells from heterogeneous populations in developing organs, which otherwise would not be available for gene analysis.

Original languageEnglish (US)
Pages (from-to)529-534
Number of pages6
JournalDevelopmental Dynamics
Volume230
Issue number3
DOIs
StatePublished - Jul 1 2004

Fingerprint

Laser Capture Microdissection
Epithelial-Mesenchymal Transition
Gene Expression
Messenger RNA
Epithelial Cells
Palate
Dermatoglyphics
Population
Genes
Real-Time Polymerase Chain Reaction
Proteins
Coloring Agents
Antibodies

Keywords

  • Gene analysis of embryonic tissues
  • LCM isolation of cells
  • Palatal mRNA during EMT

ASJC Scopus subject areas

  • Developmental Biology

Cite this

Laser capture microdissection (LCM) for analysis of gene expression in specific tissues during embryonic epithelial-mesenchymal transformation. / Nawshad, Ali; LaGamba, D.; Olsen, B. R.; Hay, Elizabeth D.

In: Developmental Dynamics, Vol. 230, No. 3, 01.07.2004, p. 529-534.

Research output: Contribution to journalArticle

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