Transfection Methods for Neurons in Primary Culture

We describe two simple, quick, and reproducible methods to transfect primary hippocampal neurons. The first method consists of a modified calcium phosphate (CP) transfection. We have developed this method to analyze localization of proteins in fully mature hippocampal neurons, however it can also be used for others purposes, such as the analysis of gene regulation or function of proteins in neurons. This method offers the advantage of transfecting small numbers of neurons at different developmental stages. The second method is an efficient electroporation (EP) protocol to perform large-scale biochemistry experiments. Examples demonstrate the suitability of both techniques. These protocols could be a useful framework to transfect primary neurons cultured from other brain regions.