OBJECTIVE: Tyro-3 and Axl receptors are expressed in brain in a region-specific manner and their bioactivities in the central nervous system remain still elusive. The aim of the present study was to investigate their functions in neuronal differentiation.METHODS: PC12 cells overexpressing Tyro-3 or Axl were established by transfection with full-length CMV-Tyro-3-eCFP or CMV-Axl-eGFP plasmid, respectively. CMV-eGFP plasmid served as a control vector. After that, the fluorescence intensity and distributions of green fluorescent protein (GFP) and cyan fluorescent protein (CFP) in the cells with or without nerve growth factor (NGF) treatment were real-time monitored.RESULTS: Expressions of Tyro-3 and Axl receptors were under the regulation of NGF and associated with neuronal differentiation. This was not observed in CMV-eGFP-transfected PC12 cells. Besides, confocal microscopy revealed that NGF affected intracellular localization of full-length Axl-eGFP and Tyro-3-eCFP in PC12 cells. Moreover, the development of outgrowth of differentiated PC12 cells under stimulation of NGF was promoted by overexpression of Tyro-3 or Axl.CONCLUSION: Expressions of Tyro-3 and Axl receptors are under the regulation of NGF and are involved in NGF-induced neuronal differentiation of PC12 cells.
Animals Blotting, Western