Membrane receptors are expressed in the plasma membrane of the cell. These molecules interact with the external environment to trigger internal subcellular processes such as signal transduction, metabolism, and other chemical processes. Because of their diverse role in physiological processes, these receptors are very important in pathological conditions such as diseases and addiction. Both External and endogenous molecules are shown to affect the expression of membrane receptors. For example, both nicotine and acetylcholine upregulate nicotinic acetylcholine receptors (nAChRs) expression. Though traditional ensemble measurement techniques provide useful information on average properties of the population, the structural and functional consequences of the heterogeneous population are hard and often impossible to be studied by such techniques. Single molecule methods, on the other hand, can elucidate powerful information about the expression, trafficking, and subunit composition of individual receptors.<br/>Nicotinic acetylcholine receptors are shown to be expressed in both pre-and post-synaptic regions of the brain. However, their expression level, subunit composition, and trafficking are not well understood. With traditional biochemical methods, it is very difficult to obtain a quantitative measurement of receptor properties and function. To address this, we have developed a total internal reflection fluorescence (TIRF) based single molecule imaging technique applicable in an <i>ex vivo</i> setting. We applied this method to study the expression of nicotinic acetylcholine receptors (nAChRs) in the synaptic regions of a mouse brain. Though some single molecule methods are very difficult to make accessible and applicable in life sciences, TIRF provides a simple way to obtain single molecule resolution by eliminating background fluorescence. Our technique can be applied to study complex pharmacological effects induced by drugs such as nicotine in synaptic regions of the brain.

Study of synaptic membrane receptors by a TIRF based single molecule method

ACS Fall 2021

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