PCOL 5-2 Cellular techniques in neuropharmacology

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32 Questions

Which type of electrode is mainly used for in vivo recordings in animals?

Extracellular electrode

What is the main disadvantage of extracellular recordings?

All of the above

Which type of electrode is mainly used in oocytes for drug screening?

Microelectrode

What is the main advantage of using microelectrodes in oocytes?

All of the above

What is the main disadvantage of using microelectrodes in neurons?

All of the above

Which of the following is true about cellular techniques in neuropharmacology electrophysiology?

It studies the electrical properties of cells or tissues

What does electrophysiology tell us?

How drugs modify the way ion channels activate

What influences cell function in relation to ion channels?

The regulation of ion channels

Which type of patch-clamping technique allows access to a whole cell and multiple ion channels?

Outside-out

What does the amplifier in patch-clamping experiments measure?

Current required to maintain membrane potential

Which type of patch-clamping technique allows access to a single ion channel in a patch of membrane?

Inside-out

What can patch-clamping techniques measure in mammalian neurons?

Voltage-dependent ion channel responses

Which patch-clamping technique allows for the control or clamping of voltage in most mammalian neurons?

Outside-out

Which type of current clamp technique measures changes in membrane potential caused by applied current?

Whole cell clamp

What does an outward current and downward deflection indicate in electrophysiology?

Positive ions leaving the cell

What is the advantage of using pulled patches in electrophysiology?

High resolution and low noise

What factors influence the size of post-synaptic current in electrophysiology?

All of the above

What can be studied using whole cell clamp technique in electrophysiology?

Intracellular milieu of cells

Which of the following is a disadvantage of patch-clamping technique?

Technically complex

What can fluorescence-based assays tell us?

How drug alters longer term processes

What is a limitation of fluorescence-based assays?

Access to inside cell

What is a disadvantage of using fluorescent materials in assays?

Interference with measurements

What is a limitation of patch-clamping technique in terms of throughput?

Low throughput

What can fluorescence-based assays measure in cells?

Membrane voltage

What is a disadvantage of fluorescence-based assays compared to electrophysiology?

Multiple readouts at once

Which of the following can be determined using immunohistochemistry?

The specific tissue components in a brain region

How can non-specific labeling be minimized in immunohistochemistry?

Blocking non-specific sites with bovine serum

What is the role of the secondary antibody in immunohistochemistry?

To interact with the primary antibody

What are the advantages of using fluorescent secondaries in immunohistochemistry?

All of the above

Which of the following is NOT a disadvantage of immunohistochemistry (IHC) technique?

Can have false negatives or false positives

What type of information does electrophysiology provide?

Function and location

Which technique allows for high throughput testing of drug action on expressed receptors?

Live cell imaging

Study Notes

Electrodes and Recordings

  • Tungsten electrodes are mainly used for in vivo recordings in animals.
  • The main disadvantage of extracellular recordings is that they are not very sensitive and can detect signals from multiple neurons.

Electrophysiology in Oocytes

  • Microelectrodes are mainly used in oocytes for drug screening.
  • The main advantage of using microelectrodes in oocytes is that they allow for high-resolution recordings of ion channels.

Microelectrodes in Neurons

  • The main disadvantage of using microelectrodes in neurons is that they can be damaged easily.

Cellular Techniques in Neuropharmacology Electrophysiology

  • Cellular techniques in neuropharmacology electrophysiology allow for the study of ion channels and their function in relation to drug action.

Electrophysiology and Ion Channels

  • Electrophysiology tells us about the electrical properties of cells and how they relate to ion channels.
  • Ion channels are influenced by the cellular environment, including pH, temperature, and ion concentrations.

Patch-Clamping Techniques

  • The whole-cell patch-clamping technique allows access to a whole cell and multiple ion channels.
  • The amplifier in patch-clamping experiments measures the tiny currents generated by ion channels.
  • The cell-attached patch-clamping technique allows access to a single ion channel in a patch of membrane.
  • Patch-clamping techniques can measure ion channel activity, membrane potential, and synaptic currents in mammalian neurons.
  • The voltage-clamp technique allows for the control or clamping of voltage in most mammalian neurons.
  • The current-clamp technique measures changes in membrane potential caused by applied current.

Electrophysiology Readings

  • An outward current and downward deflection indicate an inhibitory postsynaptic potential (IPSP) in electrophysiology.
  • Pulled patches are used in electrophysiology to increase the signal-to-noise ratio.

Whole Cell Clamp Technique

  • The whole-cell clamp technique can be used to study the properties of ion channels, synaptic currents, and neuronal excitability.

Disadvantages of Patch-Clamping Technique

  • A disadvantage of patch-clamping technique is that it is a low-throughput technique.

Fluorescence-Based Assays

  • Fluorescence-based assays can measure changes in ion channel activity, membrane potential, and synaptic currents.
  • Fluorescence-based assays can measure changes in ion channel activity in response to drug application.
  • A limitation of fluorescence-based assays is that they can be affected by photobleaching and Background fluorescence.
  • A disadvantage of using fluorescent materials in assays is that they can be toxic to cells.
  • A limitation of fluorescence-based assays compared to electrophysiology is that they are less sensitive and specific.

Immunohistochemistry

  • Immunohistochemistry can be used to determine the expression and localization of ion channels and receptors.
  • Non-specific labeling can be minimized in immunohistochemistry by using specific antibodies and blocking non-specific binding sites.
  • The secondary antibody is used in immunohistochemistry to amplify the signal and detect the primary antibody.
  • The advantages of using fluorescent secondaries in immunohistochemistry are that they allow for multiple labeling and can be used to detect multiple proteins.
  • One disadvantage of immunohistochemistry is that it can be subjective and requires expertise.

Techniques Comparison

  • Electrophysiology provides information about the electrical properties of cells and ion channels.
  • Patch-clamping technique allows for high-resolution recordings of ion channels, but it is a low-throughput technique.
  • Fluorescence-based assays can measure changes in ion channel activity, but they are less sensitive and specific than electrophysiology.
  • Immunohistochemistry can be used to determine the expression and localization of ion channels and receptors, but it can be subjective and requires expertise.

Test your knowledge on cellular techniques in neuropharmacology electrophysiology with this quiz. Learn about the electrical properties of cells and tissues, the regulation of ion channels, and the measurement of ion movement. Explore how drugs can modify these processes.

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