The sea lamprey UNC5 receptors: cDNA cloning, phylogenetic analysis and expression in reticulospinal neurons at larval and adult stages of development.

UNC5 receptors mediate repulsive signaling of netrin on neurons. Although only one UNC5 receptor has been identified in invertebrates, four members of the UNC5 family have been identified in gnathostomes. Lampreys, together with mixynes, belong to the oldest branch of extant vertebrates, and their phylogenetic position near to the vertebrate root makes them an interesting model for understanding molecular evolution. Here, we cloned three sea lamprey UNC5 (UNC5L) receptors, and phylogenetic analyses indicated that the first two duplications of the ancestral UNC5 gene occurred before the separation of jawless and jawed vertebrates. UNC5 receptors play important roles during early development, but expression studies have also suggested that UNC5 receptors play roles in the mature nervous system. Here, we report the expression of the different UNC5L receptor transcripts in identified reticulospinal neurons of mature larval or adult sea lampreys detected by in situ hybridization in wholemounted brain preparations. In addition, an extensive expression of the UNC5 receptors was also observed in most brain regions of the adult lamprey. An increase in the types of identifiable reticulospinal neurons expressing the UNC5L receptors was observed in adults compared with larvae. Expression of UNC5 receptors at late developmental stages appears to be a shared characteristic of lampreys and mammals. In larvae, expression of UNC5L receptors was observed in reticulospinal neurons that when axotomized are known to be "bad regenerators." Results in lampreys and mammals suggest that the UNC5-Netrin axonal guidance system may play a role in limiting axonal regeneration after spinal cord injury.