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Quantal analysis of inhibitory synaptic transmission by Patch clamp (CAT#: STEM-PET-0038-WXH)
Introduction
Synaptic inhibition plays an important role in sculpting signals within cortical microcircuits. At the most basic level, synaptic inhibition prevents hyperexcitability by providing activity-dependent inhibition. Inhibitory synaptic transmission uses a neurotransmitter called GABA. This interacts with GABA receptors, ion channels that are permeable to negatively charged chloride ions. Thus opening of these channels makes it harder for a neuron to generate an action potential.
Principle
The patch-clamp technique involves a glass micropipette forming a tight gigaohm seal with the cell membrane. The micropipette contains a wire bathed in an electrolytic solution to conduct ions. To measure single ion channels, a “patch” of membrane is pulled away from the cell after forming a gigaohm seal.
Applications
• Study of ionic currents in individual isolated living cells, tissue sections, or patches of cell membrane.
• Study of excitable cells such as neurons, cardiomyocytes, muscle fibers, and pancreatic beta cells.
• Study of ion channels.
• Study of excitable cells such as neurons, cardiomyocytes, muscle fibers, and pancreatic beta cells.
• Study of ion channels.
Procedure
1. Fabrication of glass electrodes
2. Measuring glass electrode resistance and compensating offset potential
3. Glass electrode contact to cell membrane and obtain a GΩ seal
4. Acquire and analyse recordings using the appropriate software.
2. Measuring glass electrode resistance and compensating offset potential
3. Glass electrode contact to cell membrane and obtain a GΩ seal
4. Acquire and analyse recordings using the appropriate software.
Materials
Patch clamp system
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