Prospects for tissue-specific analysis of gene expression in Xenopus embryos through laser-mediated microdissection of histological sections.
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.
Imamichi
Yukiko
Y
Department of Internal Medicine I, University of Ulm, Germany. yukiko.imamichi@medizin.uni-ulm.de
Koebernick
Katja
K
Wedlich
Doris
D
D000818:Animals; D017931:DNA Primers; D004210:Dissection / Q000379:methods*; D004625:Embryo, Nonmammalian; D015870:Gene Expression; D007834:Lasers; D012313:RNA / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014982:Xenopus laevis / Q000196:embryology*
D017931:DNA Primers; D012313:RNA