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Neuromuscular junctions are specialized cholinergic synapses between motor neurons and skeletal muscle cells. Understanding the mechanisms that modulate the function of these synapses are of fundamental interest. In addition, since neuromuscular synapses are critical sites for the action of acetylcholine-dependent neurotransmission, disturbance of these synapses can result in several neuromuscular diseases, such as myasthenia gravis, and can lead to an age-related decline in the strength of muscle contraction. The goal of this research is to understand how acetylcholine release is modulated at the neuromuscular junction. We have three specific aims. In Specific Aim 1 we will determine the mechanism of transmission of signals from glutamate receptors to the voltage-gated calcium channels at the nicotinic acetylcholine receptor-neural junction. In Specific Aim 2, we will study postsynaptic modulation of the voltage-gated calcium channels at the neuromuscular junction. We will determine if membrane potential affects the calcium response to acetylcholine and if the hyperpolarization-activated calcium current is modulated by activation of G protein-coupled receptors or G protein-coupled pathways such as the phosphoinositide 3-kinase pathway. In Specific Aim 3, we will determine the mechanism of postsynaptic modulation of acetylcholine release from the nerve terminal. The nicotinic acetylcholine receptor channel undergoes transition between two open states during channel activation, and the open time of the channel is regulated by receptor phosphorylation and dephosphorylation of the receptor. We will determine the mechanism by which receptor phosphorylation/dephosphorylation controls the open time of the receptor channel

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