UNIT 5.16 Green Fluorescent Protein in the Study of Neuronal Signaling Pathways
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Neuroscience
How to Cite
Blair, L., Bence-Hanulec, K. and Marshall, J. 2001. Green Fluorescent Protein in the Study of Neuronal Signaling Pathways. Current Protocols in Neuroscience. 14:5.16:5.16.1–5.16.11.
- Published Online: 1 MAY 2001
- Published Print: FEB 2001
In recent years, techniques have been established for transiently co-transfecting cells with cDNA of the jellyfish green fluoresent protein (GFP), a reporter gene that encodes a non-toxic marker. This approach can be applied to primary neurons where it has become especially useful for the study of neuronal second messenger pathways. This unit describes procedures for transfecting neurons in primary culture: transfection with GFP DNA, including co-transfecting with separate GFP and gene-of-interest constructs, transfecting with a single construct containing the gene of interest fused to a GFP gene, and transfecting with a single construct containing separate gene-of-interest and GFP cassettes. Also included is a method for the rapid, large-scale preparation of a nearly homogeneous population of neurons from rat cerebellum. The Commentary provides several examples of how this approach can be applied to specific biological questions on neuronal signaling pathways.