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Related Concept Videos

Patch Clamp01:18

Patch Clamp

7.0K
Many fundamental cell functions such as muscle contraction and nerve transmission rely on the electrical signals produced by the movement of positively and negatively charged ions across the cell membrane. One competent method to record current flowing across the whole cell or single ion channel is the patch-clamp technique.
In this method, a glass micropipette containing electrolyte solution is tightly sealed against a small portion of the cell membrane. As a result, a patch of the cell...
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Cholinergic Receptors: Nicotinic01:15

Cholinergic Receptors: Nicotinic

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Nicotinic receptors are ligand-gated ion channels that are activated by acetylcholine and nicotine. Upon activation, they cause a rapid increase in the permeability of cells to K+, Na+, and Ca2+, followed by depolarization and excitation. They are in the autonomic ganglia, skeletal neuromuscular junction, CNS, and adrenal medulla.
There are two types of nicotinic receptors: neuromuscular (NM/NM/N1) and neuronal (NN/NN/N2). The two families differ based on their location and selectivity to...
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Cholinergic Receptors: Muscarinic01:25

Cholinergic Receptors: Muscarinic

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The pharmacological actions of acetylcholine are elicited via its binding to two families of cholinergic receptors or cholinoceptors, namely, muscarinic and nicotinic receptors. Muscarinic receptors are G protein-coupled receptors and have five subtypes, M1–M5. All mAChR subtypes are activated by acetylcholine and blocked by the antagonist, atropine. 
The subtypes M1, M3, and M5 couple with the Gq subunit and activate the phospholipase C (PLC) activity, mobilizing intracellular Ca2+....
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Related Experiment Video

Updated: Mar 6, 2026

Local Application of Drugs to Study Nicotinic Acetylcholine Receptor Function in Mouse Brain Slices
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Local Application of Drugs to Study Nicotinic Acetylcholine Receptor Function in Mouse Brain Slices

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High-Throughput Patch Clamp Screening in Human α6-Containing Nicotinic Acetylcholine Receptors.

Lucas C Armstrong1, Glenn E Kirsch1, Nikolai B Fedorov1

  • 11 Charles River Discovery, Cleveland, OH, USA.

SLAS Discovery : Advancing Life Sciences R & D
|March 17, 2017
PubMed
Summary

Researchers developed a new cell line to study specific nicotinic acetylcholine receptors (nAChRs) involved in nicotine addiction. This tool helps identify compounds that selectively target these receptors, advancing addiction research.

Keywords:
automated patch clampelectrophysiological screeningion channelnicotinic acetylcholine receptor

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Spectral Confocal Imaging of Fluorescently tagged Nicotinic Receptors in Knock-in Mice with Chronic Nicotine Administration
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Area of Science:

  • Neuroscience
  • Pharmacology
  • Molecular Biology

Background:

  • Nicotine, the primary addictive substance in tobacco, acts on nicotinic acetylcholine receptors (nAChRs) in the brain.
  • The α6β2β3 nAChR subtype, found in dopaminergic neurons, plays a crucial role in modulating dopamine release within brain regions implicated in nicotine addiction.
  • Understanding the subtype-selective actions of tobacco constituents on nAChRs is vital for elucidating the neurological basis of tobacco addiction.

Purpose of the Study:

  • To develop and validate a novel recombinant cell line expressing the human α6/3β2β3V273S nAChR subtype.
  • To establish a robust screening and profiling assay using an automated patch clamp platform for this specific nAChR subtype.
  • To identify novel modulators of the α6β2β3 nAChR subtype.

Main Methods:

  • Development of a recombinant cell line engineered to express the human α6/3β2β3V273S nAChR.
  • Pharmacological characterization of the cell line using selective agonists, antagonists, and pore blockers.
  • Utilization of an automated patch clamp system (IonWorks Barracuda) for high-throughput screening and electrophysiological assays.
  • Pilot screening of a library of Food and Drug Administration-approved drugs.

Main Results:

  • The developed cell line and automated patch clamp assays accurately replicated findings from conventional electrophysiological methods.
  • Pharmacological characterization confirmed the specificity and functionality of the expressed α6/3β2β3V273S nAChR.
  • A pilot screen identified previously unrecognized FDA-approved drugs that selectively inhibit the α6/3β2β3V273S nAChR subtype.

Conclusions:

  • The novel recombinant cell line and automated patch clamp assays provide effective tools for screening and subtype-selective profiling of compounds targeting α6β2β3 nicotinic receptors.
  • These tools can accelerate the discovery of novel therapeutic agents for nicotine addiction and related neurological disorders.
  • The findings highlight the potential of existing drugs to modulate specific nAChR subtypes, opening new avenues for drug repurposing.