A novel seven transmembrane receptor induced during the early steps of astrocyte differentiation identified by differential expression

The rat glial progenitor cell line CG-4 can be induced to differentiate into either oligodendrocytes or type-2 astrocytes. In order to identify genes whose expression varies coincident with such phenotypic differentiation, we employed representational difference analysis (RDA) of mRNA. Here, we report 38 cDNAs induced in type-2 astrocytes, oligodendrocytes, or both differentiated states. Among these were known transcription factors, membrane receptors, extracellular matrix proteins, secreted signaling modulators, chromatin regulators and myelin sheath components. In addition several novel genes were identified; among these was a gene induced during the very early stages of astrocyte differentiation that we have named leda (induced early in differentiating astrocytes). Several leda transcripts were detected by RT-PCR, and appeared to be produced by alternative splicing and promoter usage. The protein deduced from the longest leda mRNA exhibited sequence features characteristic of G-protein coupled receptors, including seven putative transmembrane domains, while the shorter leda transcripts encoded proteins that lacked several transmembrane segments. In the adult rat, leda transcripts were found exclusively in brain and testis. In the developing rat brain, leda expression was first detected at embryonic day 16, that is two days before the first appearance of mature astrocytes. Thus, this approach has yielded a potential source of markers for differentiation states of these two cellular types as well as genes predicted to be functionally involved in the differentiation process itself.