Whole-cell patch-clamp recording and parameters.

Biophys Rev

Cardiovascular Division, Brigham and Women's Hospital, Harvard Medical School, Boston, USA.

Published: April 2023

AI Article Synopsis

  • - The patch-clamp technique is an advanced electrophysiological method used to study single channels in cell membranes, with five main configurations (loose patch, cell-attached, whole-cell, inside-out, outside-out) to explore various electrical signals like action potentials and resting membrane potential.
  • - This method allows for the detailed analysis of ion channel behavior, indicating that even small malfunctions can significantly affect action potentials, and it provides reliable recordings if performed correctly.
  • - While patch-clamp is a powerful tool for examining ion currents, other methods like optical techniques exist but may not yield as robust signals, demonstrating the complexity behind cellular electrophysiology and its evolutionary similarities across species.

Article Abstract

The patch-clamp technique represents an electrophysiology type of method. This is one of several insightful approaches with five major configurations, namely a loose patch, cell-attached (also known as on-cell), whole-cell, inside-out, and outside-out modes. The patch-clamp method is more advanced compared to classical electrophysiology since it elucidates single-channel activation in a tiny portion of the membrane in addition to action potential (AP), junction potential (JP), endplate potential (EP), electrical coupling between two adjacent cells via Gap junction hemi-channels, excitatory/inhibitory postsynaptic potentials, and resting membrane potential (RMP). In fact, a malfunction of only one channel or even one component will alter AP amplitude or duration in vitro. If parameters are inferred appropriately and recordings are performed properly, the patch-clamp trace readouts and results are robust. The main hallmarks of currents via voltage-dependent calcium (Cav), hyperpolarization-activated cyclic nucleotide gated non-selective cation (HCN), inwardly rectifying potassium (Kir), voltage-dependent potassium (Kv), and voltage-dependent sodium (Nav) channels are similar and tractable among cells even when they are derived from evolutionary distinct organs and species. However, the size of the membrane area, where the functional subunits reside, and current magnitudes vary among cells of the same type. Therefore, dividing current magnitudes by cell capacitance- current density enables the estimate of functional and active channels relative to recorded cytoplasmic membrane area. Since the patch-clamp recordings can be performed in both current- and voltage-clamp modes, the action potential or spike durations can be adequately elucidated. Sometimes, optical methods are preferred to patch-clamp electrophysiology, but the obtained signals and traces are not robust. Finally, not only an alternans of AP durations, but also that of 'action potential shape' is observed with electrophysiology.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10133435PMC
http://dx.doi.org/10.1007/s12551-023-01055-8DOI Listing

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