mRNA expression profiling of laser microbeam microdissected cells from slender embryonic structures.
Microarray hybridization has rapidly evolved as an important tool for genomic studies and studies of gene regulation at the transcriptome level. Expression profiles from homogenous samples such as yeast and mammalian cell cultures are currently extending our understanding of biology, whereas analyses of multicellular organisms are more difficult because of tissue complexity. The combination of laser microdissection, RNA amplification, and microarray hybridization has the potential to provide expression profiles from selected populations of cells in vivo. In this article, we present and evaluate an experimental procedure for global gene expression analysis of slender embryonic structures using laser microbeam microdissection and laser pressure catapulting. As a proof of principle, expression profiles from 1000 cells in the mouse embryonic (E9.5) dorsal aorta were generated and compared with profiles for captured mesenchymal cells located one cell diameter further away from the aortic lumen. A number of genes were overexpressed in the aorta, including 11 previously known markers for blood vessels. Among the blood vessel markers were endoglin, tie-2, PDGFB, and integrin-beta1, that are important regulators of blood vessel formation. This demonstrates that microarray analysis of laser microbeam micro-dissected cells is sufficiently sensitive for identifying genes with regulative functions.
Scheidl
Stefan J
SJ
Department of Medical Biochemistry, Göteborg University, Göteborg, Sweden.
Nilsson
Sven
S
Kalén
Mattias
M
Hellström
Mats
M
Takemoto
Minoru
M
Håkansson
Joakim
J
Lindahl
Per
P
D000818:Animals; D004622:Embryo, Mammalian / Q000166:cytology / Q000502:physiology*; D005260:Female; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D007834:Lasers; D051379:Mice; D008810:Mice, Inbred C57BL; D008846:Micromanipulation; D020411:Oligonucleotide Array Sequence Analysis; D011247:Pregnancy; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics
D012333:RNA, Messenger