Molecular Cloning and In Situ Localization in the Brain of Rat Sky Receptor Tyrosine Kinase

Abstract

We previously identified a human cDNA encoding a novel receptor tyrosine kinase, termed Sky, which is predominantly expressed in the brain and has a unique extracellular domain consisting of two immunoglobulin (1g)-like and two fibronectin type III (FN III) motifs. In attempts to define the functional role of the Sky receptor, we cloned a rat sky cDNA, and localized the sites of expression of sky transcript in the adult rat brain by Northern blot and in situ hybridization analyses using the cloned rat cDNA as a probe. The deduced amino acid sequence of rat Sky has an overall sequence and a domain topology highly conserved with human Sky (90% overall identity and 98% identity within the tyrosine kinase domain). Northern blot analysis revealed that a single 3.8-kb sky mRNA is expressed in PC12 pheochromocytoma and Neuro-2a neuroblastoma cell lines and in various regions of the adult rat brain. In situ hybridization analysis revealed widespread but confined neuronal populations in adult rat brain that express sky transcript; prominent hybridization signals were detected in the inner granular layer of the olfactory bulb, CA-1 area of the hippocampus, granule cell layer of the cerebellum, tenia tectum and cingulate gyrus neurons, and wide regions of cortex layers II-VI. The high level of expression of sky mRNA in neurons in restricted brain regions suggests that the Sky receptor may play an important role in development, function, and maintenance of specific neuronal populations in the central nervous system.