Journal Article

Prospects for Tissue-specific Analysis of Gene Expression in Xenopus Embryos through Laser-mediated Microdissection of Histological Sections

Wedlich, Doris, Koebernick, Katja, Imamichi, Yukiko

2003

Overview

Analysis of spatiotemporal patterns of gene expression is an important prerequisite for understanding the molecular basis of embryogenesis. Tissue-specific resolution is desirable, but often not achieved owing to methodical limitations. We used a common model system for embryonic development – the South African clawed frog Xenopus laevis – to demonstrate that laser microdissection and laser-mediated catapulting of tissue samples from histologic sections are feasible even for yolk-rich, fragile embryonic tissue. A combination with RT-PCR provides the possibility of detecting tissue-specific gene expression with high resolution and fidelity. We show that specimens of various sizes and shapes can easily be procured by laser microdissection and pressure catapulting (LMPC). Subsequent RNA-isolation and nested RT-PCR for marker genes revealed that the combination of these methods allows for analysis of specific gene expression in micro-areas. We report on the efficiency and reliability of detection of marker genes in dissected tissue. We further discuss the question of whether such a combination can be applied to certain issues raised in developmental biology with regard to other techniques.

• Publisher: Elsevier GmbH,Elsevier Science Ltd
• Subject: Animals / Developmental biology / Dissection - methods / DNA Primers / Embryo, Nonmammalian / Gene Expression / Laser mediated catapulting (LMC) / Laser microbeam microdissection (LMM) / Lasers / Reverse Transcriptase Polymerase Chain Reaction / RNA - analysis / RT-PCR / Xenopus laevis / Xenopus laevis - embryology