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Related Concept Videos

The Resting Membrane Potential01:21

The Resting Membrane Potential

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Potentiometry: Membrane Electrodes01:15

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Membrane electrodes, also known as p-ion electrodes, use membranes that selectively interact with free analyte ions, generating a potential difference across the membrane. The resulting membrane potential, known as the asymmetry potential, is not zero even when analyte concentrations on both sides of the membrane are equal. The membrane's response is typically not selective to a single analyte but proportional to the concentration of all ions in the sample solution capable of interacting at the...
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The relative difference in electrical charge, or voltage, between the inside and the outside of a cell membrane, is called the membrane potential. It is generated by differences in permeability of the membrane to various ions and the concentrations of these ions across the membrane.
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Resting Membrane Potential01:24

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The resting membrane potential of a neuron (-70mV) is sustained due to the selective ion permeability of the membrane. At the resting potential, the membrane is slightly permeable to ions like sodium (Na+) and chloride (Cl−) and highly permeable to potassium ions (K+). Differences in the ions' concentration inside the cell compared to the outside are maintained by membrane transport proteins like channels and pumps.
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Potentiometry is an analytical technique that measures the potential difference between two electrodes in an electrochemical cell without drawing any significant current that could alter the solution's composition. This method employs an indicator electrode, which exchanges electrons with the analyte solution, and a reference electrode with a constant potential. Each electrode is immersed in a solution comprised of two half-cells. In a conventional setup, the reference electrode serves as the...

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Measuring the Induced Membrane Voltage with Di-8-ANEPPS
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Quantitative analysis of membrane potentials.

Manus W Ward1

  • 1Department of Physiology and Medical Physics, Royal College of Surgeons in Ireland, Dublin, Ireland.

Methods in Molecular Biology (Clifton, N.J.)
|December 4, 2009
PubMed
Summary
This summary is machine-generated.

Researchers can now quantitatively assess mitochondrial energetics and membrane potential changes using potentiometric fluorescent probes. Accounting for plasma membrane potential is crucial for accurate mitochondrial assessments, aided by computational modeling.

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Area of Science:

  • Cellular and Molecular Physiology
  • Biophysics
  • Biotechnology

Background:

  • Electrochemical gradients across biological membranes are key indicators of physiological and pathophysiological processes.
  • Accurate measurement of mitochondrial membrane potential (Δψm) is essential for understanding cellular energetics and drug effects.
  • Cationic fluorescent probes (e.g., TMRM, TMRE, Rhodamine 123) are commonly used to assess Δψm but their interpretation is complicated by concurrent changes in plasma membrane potential (Δψp).

Purpose of the Study:

  • To provide researchers with methods for quantitative assessment of mitochondrial energetics at a single-cell level.
  • To detail the use of potentiometric fluorescent probes for measuring mitochondrial membrane potential (Δψm).
  • To address the challenge of interpreting Δψm measurements by incorporating plasma membrane potential (Δψp) assessments and computational modeling.

Main Methods:

  • Utilizing potentiometric fluorescent probes (TMRM, TMRE, Rhodamine 123) sensitive to mitochondrial membrane potential (Δψm).
  • Employing anionic fluorescent probes to monitor changes in plasma membrane potential (Δψp) for validation.
  • Developing and applying computational modeling techniques based on probe redistribution to analyze simultaneous Δψm and Δψp changes.

Main Results:

  • Demonstrated a quantitative approach to assess mitochondrial energetics via Δψm measurements using cationic fluorescent probes.
  • Highlighted the necessity of accounting for Δψp to accurately interpret Δψm probe responses.
  • Showcased computational modeling as a tool to resolve complex changes in both Δψm and Δψp.

Conclusions:

  • Accurate single-cell assessment of mitochondrial energetics requires simultaneous measurement and analysis of both mitochondrial (Δψm) and plasma (Δψp) membrane potentials.
  • Potentiometric fluorescent probes, when used in conjunction with computational modeling, offer a robust method for quantitative analysis of cellular electrochemical gradients.
  • This approach enhances the understanding of cellular responses, disease mechanisms, and drug interactions.