Equipment for brain slice patch-clamp electrophysiology

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Communication between neurons in the brain is governed by synaptic transmission, which is mediated by the presynaptic release of chemical neurotransmitters and their binding to cell surface receptors on the postsynaptic neuron. The research in our lab focuses on a class of ligand-gated ion channels called nicotinic acetylcholine receptors. Our long term goal is to examine the basic function of nicotinic receptors in modulating neurotransmission in the CNS. We have developed a novel approach to understanding nicotinic receptor mediated signaling in the brain. We have genetically engineered knock-in mice in which we fused a fluorescent protein to the alpha4 nicotinic receptor subunit. To our knowledge this is the first knock-in mouse in which an ion channel has been tagged with a fluorescent protein. Alpha4 beta2 nicotinic receptors constitute the major nicotinic receptor subtype in the CNS. For the first time, we have the ability to quantify accurately at submicron resolution the subcellular distribution of the receptor in neurons in the CNS. This important information will allow us to precisely target those neurons for electrophysiology to ascertain the function of alpha4 subunit containing nicotinic receptors. Furthermore, we have also generated knock-in mice with a gain-of-function mutation to the alpha4 nicotinic receptor gene. This mutation renders the receptor 50 fold more sensitive to activation by nicotine or acetylcholine and expresses four fold greater nicotinic responses thus facilitating the recording of alpha4 nicotinic receptors.