Nucleofection of cultured hippocampal neurons

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In order to understand how the brain functions we must be able to record the electrical activity occurring in specialized brain cells called neurons. In order to make electrical recordings from these neurons we need to remove them from the mammalian brain. To keep the neurons alive outside of the brain, they are cultured in an incubator for up to three weeks. The first piece of equipment we are requesting is a laminar flow hood, which is used in the culturing process. Without this piece of equipment the neurons become infected with bacteria and viruses found in the environment; once they are infected they can no longer be used for experiments. Following successful culturing, we need to alter the expression of proteins of interest in order to determine their function. To do this we need to transfect neurons with DNA and RNA that code for the protein we are interested in examining. It is difficult to transfect neurons properly and maintain cell health. However, there is a relatively new piece of equipment called the Amaxa Nucleofector, which achieves high-efficiency stable transfection with no compromise in cell health. We are requesting funds to purchase an Amaxa Nucleofector to allow our investigation of the cellular and molecular mechanisms underlying inhibitory synaptic plasticity. This research will benefit Canadians who suffer from memory problems associated with normal aging and neurological disorders, including epilepsy and Alzheimers'.