THE NONRECEPTOR TYROSINE KINASE LYN IS LOCALIZED IN THE DEVELOPING MURINE BLOOD-BRAIN-BARRIER

The blood-brain barrier, formed by brain endothelium, is critical for brain function. The development of the blood-brain barrier involves brain angiogenesis and endothelial cell differentiation, processes which require active signal transduction pathways. The differentiation of brain endothelial cells to the ''blood-brain-barrier phenotype'' involves cytoskeletal changes which modulate the tightness of the barrier. In order to identify signal transduction proteins involved in blood-brain barrier development, cDNA from bovine and murine brain endothelial cells was used in a polymerase chain reaction for cloning of DNA encoding Src homology 3 domains. Src homology 3 domains are structural domains found in many signal transduction proteins. These domains often mediate interaction of signaling proteins with the cytoskeleton and therefore may play a role in the regulation of the cytoskeletal changes which occur during blood-brain-barrier development. Unexpectedly, all bovine and murine clones analyzed from polymerase chain reactions encoded the Src homology 3 domain of one protein, namely the non-receptor tyrosine kinase, Lyn, which is involved in signal transduction in cells of the hemopoietic system. In situ hybridization analyses confirmed the presence of lyn mRNA in developing blood vessels in embryonic and early post-natal mouse brain, but not in endothelium outside the brain. In bovine brain endothelial cells in primary culture, p53(lyn) is highly abundant and present in two forms which have different patterns of tyrosine phosphorylation, These data suggest that Lyn may be involved in transduction of growth and differentiation signals required for blood-brain-barrier development.