Renal transcriptomes: segmental analysis of differential expression.
OBJECTIVE
Progress accomplished by complete genomes and cDNA-sequencing projects calls for methods that fully use these resources to study gene expression patterns in characterized cell populations. However, since the number of functional genes cannot be readily inferred from the genomic sequence, it is highly desirable to make use of methods enabling to study both known and unknown genes.


METHODS
The method of serial analysis of gene expression provides short diagnostic cDNA tags without bias towards known genes. In addition, the frequency of each tag in the library conveys quantitative information on gene expression. A microassay was set-up to perform serial analysis of gene expression in minute samples such as those obtained by microdissecting nephron segments.


RESULTS
Studies carried out in the thick ascending limb of Henle's loop and the collecting duct of the mouse kidney provided expression data for several thousand genes. Known markers were found appropriately enriched, and several of the thick ascending limb or collecting duct specific transcripts had no database match.


CONCLUSIONS
The microassay for serial analysis of gene expression makes possible large-scale quantitative measurements of mRNA levels in nephron segments. The comprehensive picture generated by analyzing both known and unknown transcripts in defined cell populations should help to discover genes with dedicated functions.
Elalouf
Jean-Marc
JM
Département de Biologie Cellulaire et Moléculaire, Service de Biologie Cellulaire, CNRS URA 1859, CEA SACLAY, F-91191 Gif-sur-Yvette, France. elalouf@dsvidf.cea.fr
Aude
Jean-Christophe
JC
Billon
Emmanuelle
E
Cheval
Lydie
L
Doucet
Alain
A
Virlon
Bérangère
B
D000818:Animals; D020869:Gene Expression Profiling; D023281:Genomics; D006801:Humans; D007668:Kidney / Q000502:physiology*; D014158:Transcription, Genetic / Q000235:genetics*