Characterization of progenitor-cell-specific genes identified by subtractive suppression hybridization

We have utilized subtractive suppression hybridization (SSH) to identify differentially expressed genes present in either neuroepithelial (NEP) cells or glial restricted precursor (GRP) cells. Eighteen clones enriched in GRP cells and 28 in NEP cells were identified. Five of the GRP-specific clones (tenascin C, cystatin C, GABA transporter 3, extracellular matrix molecule 2 and H2-4) were characterized further, and their glial specificity was confirmed by RT-PCR, in situ hybridization and immunocytochemistry. H2-4 (an expressed sequence tag) was shown to be part of chondroitin sulfate proteoglycan 3. Overall, our results show that SSH can be used to identify lineage-and stage-specific markers and that extracellular matrix molecules likely play important roles in the migration and differentiation of GRPs.