Introductory Medical Physiology

Sept., 2011 Prof. Dr. Hamdan Noor

i. Overview
Resting membrane potential cannot carry any signal unless and until it is transformed to graded potential and then to action potential. Only action potential can be propagated to carry the message about a stimulus so that appropriate response can be elicited in the target cell. A stimulus triggers transformation of resting membrane potential at special sites of an excitable tissue (at a receptor or sensor, post-synaptic membrane, endplate surface of skeletal muscle, or pacemaker) to produce graded potential. This is due to stimulation of chemical-gated or mechanicalgated ion channels that open and cause a change the permeability of the membrane to certain ions. If the stimulus produces a graded potential that depolarises enough to reach threshold, this voltage difference will stimulate voltage-gated ion channels located close to the receptor site. The membrane will depolarize to form an action potential. This depolarization will in turn stimulate the next voltage-gated channel along the axon and another action potential is produced. This process continues to occur until the action potential reaches the axon terminal, thus carrying the message about the stimulus to be communicated to the next neuron or effector cell.

ii. Content
1. Membrane potentials as information signal 1.1. The role of the nervous system in intercellular communication 1.2. From resting membrane potential to information signal 1.3. Membrane potentials and ion channels 1.4. Locations of the leakage and gated channels and their implication Change in membrane potentials when stimulated: Graded potential 2.1. From resting potential to graded potential 2.2. Graphical representation of graded potential 2.3. Analysis of graded potential Concept of threshold Change in membrane potentials when stimulated: Action potential Conduction/propagation/transmission of action potential 5.1. Mechanism of action potential propagation 5.2. The significance of refractory periods 5.3. Strategies to hasten action potential propagation

2.

3. 4. 5.

iii. Checklist of topics and learning activities in this module

Content i ii iii iv v vi 1 Abstract Learning Resources Background Knowledge Terms to Know Objectives Learning Activities Membrane potentials as information signal 1.1. The role of the nervous system in intercellular communication ACTIVITY 7.1: Significance of intercellular communication 1.2. From resting membrane potential to information signal ACTIVITY 7.2: Membrane potential as information signal 1.3. Membrane potentials and ion channels ACTIVITY 7.3. Significance of ion channels ACTIVITY 7.4: Implication of ionic diffusion via gated channels 1.4. Locations of the leakage and gated channels and their implication ACTIVITY 7.5. Locations of membrane channels Change in membrane potentials when stimulated: Graded potential 2.1. From resting potential to graded potential ACTIVITY 7.6: Equilibrium potential ACTIVITY 7.7: Demonstration on change in membrane potential ACTIVITY 7.8: Demonstration of depolarisation ACTIVITY 7.9: Graded potential 2.2. Graphical representation of graded potential ACTIVITY 7.10: Analysing membrane potential graphically 2.3. Analysis of graded potential ACTIVITY 7.11: Molecular mechanism of graded potential ACTIVITY 7.12: Types of graded potentials ACTIVITY 7.13: EPSP and IPSP ACTIVITY 7.14: Graphs of graded potential Page Comments on mastery

Concept of threshold ACTIVITY 7.15: Stimulus-response relationship 4 Change in membrane potentials when stimulated: Action potential ACTIVITY 7.16: Action potential ACTIVITY 7.17: Mechanism of action potential production 5 Conduction/propagation/transmission of action potential 5.1. Mechanism of action potential propagation ACTIVITY 7.18: Mechanism of action potential propagation ACTIVITY 7.19: The domino effect of action potential propagation 5.2. The significance of refractory periods ACTIVITY 7.20: Relevance of refractory periods 5.3. Strategies to hasten action potential propagation ACTIVITY 7.21. Saltatory conduction of action potential vii Summary viii Conclusion ix Assessment

iv.

Learning Resources

Please refer to the following sources for further information: Boron and Boulpaep, Ch. 2 & 3 Guyton & Hall, pp 10-12 & Ch. 4 Ganong, Ch. 1 Marieb, Ch. 3. + Study partner Tortora & Grabowski, Ch. 3 Vander, Sherman & Luciano, Ch. 3 Supplementary materials provided

Animations: 1. http://bcs.whfreeman.com/thelifewire/content/chp44/4402s.swf tutorial on action potential 2. http://www.tvdsb.on.ca/westmin/science/sbioac/homeo/action.htm action potential 3. http://www.accessexcellence.org/RC/VL/GG/action_Potent.html Propagation of action potentials 4. http://outreach.mcb.harvard.edu/animations/actionpotential.swf action potential

v. Background knowledge
To complete this module successfully, you should have the following background:

Knowledge on the general structure and functions of cell membranes (Module 2). Knowledge on the mechanisms of solute transport across cell membrane (Module 3). Knowledge on the basic mechanism of signal transduction (Module 4). Understanding of the definition of chemical and electrical gradients (Module 5). Knowledge on membrane potentials of excitable tissues at rest (Module 5).

vi. Terms to know

After studying the materials and doing the activities in this module, students should be comfortable with the following terms: action potential active transport body fluid compartments channels chemically activated gates concentration concentration gradients conductance cytoplasm depolarization diffusion dynamic equilibrium electrical gradients electrode electrochemical gradient electrogenic pump excitatory post-synaptic potential (EPSP) fatty acid (tails) gated channels hydrophilic interstitial fluid (ISF) hydrophobic hyperpolarized inhibitory post-synaptic potential (IPSP) ion binding sites membrane permeability membrane potentials milliseconds (msec). Na + /K + -ATPase net charge net movement passive channels phospholipid bilayer receptor site resting membrane potential semi-permeable solute solvent threshold voltage activated gates voltmeter

Please add other terms that you feel are relevant to your understanding of this module.

vii. Learning objectives

After studying the materials in Module 6, the students should be able to: 1. Discuss the role of the nervous system in intercellular communication. 2. Discuss the sequence of events that take place on a cell membrane when a stimulus reaches a receptor, resulting in a change in membrane permeability to certain ions, consequently changing the membrane potential. 3. Describe the significance of the presence of different ion channels in the membrane of a neuron in terms of their effects on membrane potential. 4. Define graded potential, describe how it is established, and describe its characteristics. List down the types of graded potential (including EPSP and IPSP) and state the significance of each. 5. Plot the membrane potential response to subthreshold, threshold, and suprathreshold stimuli and explain the molecular basis of the response. Discuss their significance. 6. Explain the meaning of action potential; describe the mechanism of action potential production and action potential propagation. 7. Compare and contrast between action potential and graded potential. 8. Describe the molecular basis of absolute and relative refractory periods and state the significance. 9. Describe the strategies that can hasten action potential propagation.

Please set up more specific objectives after youve thoroughly studied the material in this module to help yourself in your revision later on. Make notes that meet the requirement of the new objectives.

Objectives from the American Physiological Society

CE 10. Based on the principle of ionic attraction, explain how a potential difference across a membrane will influence the distribution of a cation and an anion. CE 11. Define the term steady state, and differentiate it from equilibrium. Relate the pump-leak model of steady-state ion content to cell solute gradients and cell volume maintenance. CE 12. Write the Nernst equation, and indicate how this equation accounts for both the chemical and electrical driving forces that act on an ion. CE 13. Based on the Nernst equilibrium potential, predict the direction that an ion will move when the membrane potential a) is at its equilibrium potential, b) is higher than the equilibrium potential, or c) is less than the equilibrium potential. List values in a typical non-excitable cell for the membrane potential, for ENa, EK, ECl, and ECa. CE 14. Define the concepts of electrochemical equilibrium and equilibrium potential, and give internal and external ion concentrations. Be able to calculate an equilibrium potential for that ion using the Nernst equation. Contrast the difference in EK (the Nernst potential for K+) caused by a 5 mEq/l increase in extracellular K+ with the change in ENa (the Nernst potential for Na+) caused by a 5 mEq/l increase in extracellular Na+. CE 15. Explain how the resting membrane potential is generated and calculate membrane potential by using either a) the Goldman-Hodgkin-Katz equation or b) the chord conductance equation. Given an increase or decrease in the permeability of K, Na, or Cl, predict how the membrane potential would change.

viii. Learning activities

1. Membrane potentials as information signal
In Module 6 we learned about the membrane potential when the cell is not stimulated. We know that resting membrane potential exists in every cell, but resting potential does not carry any message even in excitable cells, as there is no stimulation. Nevertheless, when the cell is stimulated, the resting membrane potential is transformed into action potential by excitable cells (non-excitable cells are not able to do this). Action potential can be propagated along the excitable cell to affect adjacent cells. In this Module, well explore how excitable cells transform resting membrane potential in the presence of appropriate stimulus. This forms the basis of intercellular communication via the nervous system. 1.1. The role of the nervous system in intercellular communication You have learnt about intercellular communication in Module 5. Activity 7.1 helps you to review the significance of intercellular communication in body functions.
ACTIVITY 7.1: Significance of intercellular communication Human beings must fulfill two basic physiological needs: a) to regulate chemical and physical factors in the internal environment, and b) to respond/adapt to external environment. How do we regulate the chemical and physical factors in the internal environment? What are the basic components of homeostasis? Give examples of homeostatic regulation, and name all the specific components. Explain the mechanism of information flow from one component to another. Why do we need to respond to the external environment? Give examples. Explain the processes (information flow) that are involved in the perception of the external environment. Use Fig. 7.1 to help you in your answer. From the above explanation, deduce the significance of intercellular communication. Receptor Control centre

Suggest some characteristics of response pathways that involve changes in internal environmental parameters vs external environmental parameters. Hint: receptor location, control centre location (perception), effector type, and general response (reflex).

Effector

Figure 7.1. Information pathway from receptor to effector

1.2. From resting membrane potential to information signal From Activity 7.1 you infer that intercellular communication may occur via the nervous system. You also know that neurons are excitable i.e. it can generate impulses that are used as information signals. However, we know from Module 6 that resting membrane potential does not carry any message (why?). Therefore, the resting potential must be transformed in such a manner so as to function as signal carrier. Activity 7.2 helps you to predict the sequence of events that take place on a cell membrane when a stimulus reaches a receptor.
ACTIVITY 7.2: Membrane potential as information signal How is resting membrane potential transformed so that it can be used to transmit information? Hint: 1. Firstly, there must be a stimulus (give examples of internal and external stimuli) to trigger the change. 2. Secondly, there must be an appropriate receptor to the stimulus that is associated with ion channels. 3. Thirdly, the stimulus interacts with membrane receptors change the configuration of ion channels on the membrane change the permeability of membrane to certain ions by altering the characteristics of channel proteins change the membrane potential. Predict the consequence of the above events. Hint: Change in the pattern of ion movement across the membrane. What happens to the membrane potential if the ion channels involved + are Na channels? Predict the implication of the above events. Hint: Resting potential is changed to some other potentials that can be used to carry information (Refer to Table 7.1).

The key molecules that are involved in producing information signal on a cell membrane are the proteins such as stimulus receptors and channel proteins. Before we look at what happens to the membrane potential when stimulated, lets review the relationship between ion channels and membrane potentials.

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1.3. Membrane potentials and ion channels We know from Module 6 that: ions move across the membrane through ion channels. channels are specific for each ion. membrane potential is the result of unequal distribution of ions across the plasma membrane. We also know that there are two main types of ion channels (Fig 7.2): Passive, or leakage channels that are always open. Active, or gated channels that open or close only in response to various signals. There are three main types of gated channels: o Chemical-gated channel o Voltage-gated channel o Mechanical-gated channel
Ion channels

Passive (leakage)

Active (gated)

Chemicalgated

Voltagegated

Mechanicallygated

Fig. 7.2. Types of ion channels

Activity 7.3 helps you to understand the significance of ion channels.

ACTIVITY 7.3: Significance of ion channels What is the implication of the number of leakage K and Na channels in plasma membrane to the resting potential? You have calculated the EK+ and ENa+ using the Nernsts equation, and the EM using the Goldmans equation. Why is the resting + + membrane potential closer to the equilibrium potential of K compared to that of Na ? Predict what happens to a cell that has only leakage channels (no gated channels) when it is stimulated by an appropriate stimulus. Which type of cell has no gated channels? Which type of cell has gated channels? What is the advantage of having gated channels? Why dont all cells have gated channels? What kind of gated channels exist in receptor cells that directly respond to internal and external environmental stimuli?
+ +

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Ions move along chemical and electrical gradients (electrochemical gradients) through the channels, carrying the +ve or ve charges. This will cause a change in membrane potential. Activity 7.4 helps you to clarify this phenomenon.

Fig. 7.3. Chemical-gated channel

Fig. 7.4. Voltage-gated channel.

ACTIVITY 7.4: Implication of ionic diffusion via gated channels Refer to Fig. 7.3: What happens when acetylcholine binds to its receptor in the membrane? Draw a graph to show resting membrane potential and the change in potential when acetylcholine binds to the receptor. Refer to Fig. 7.4: + What happens to the voltage gated K channel when the membrane potential is changed from -70 mV to -50 mV? What is the implication in terms of the resulting membrane potential? Draw a graph to show resting membrane potential and the change in potential + when the voltage-gated K channel is stimulated by an initial depolarization.

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1.4. Locations of the leakage and gated channels and their implication Leakage channels are found almost everywhere on a cell membrane. However, the number of individual channels on a cell membrane may be different (refer to Activity 7.8). Chemical/mechanical-gated channels are located on: nerve endings or receptor (sensor) sites post-synaptic membrane pacemaker cells muscle (neuromuscular junction) Voltage gated channels are located at axon hillock and along the axon distal to receptor region (Fig. 7.5)
ACTIVITY 7.5: Locations of membrane channels List down the locations of leakage channels, chemicalgated and mechanical gated channels in cell membrane. Hypothesise the significance of the locations of the leakage and gated channels. Based on Fig. 7.5 locate a\and suggest the function of: Input zone, Trigger zone, Conducting zone, Output zone What ion channels are present at the above zones? What are the implications?

Axon hillock

Figure 7.5. Nerve endings or receptor sites have gated channels that are activated by appropriate stimuli.

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2. Change in membrane potentials when stimulated: Graded potential

2.1. From resting potential to graded potential In Module 6 you have worked on equilibrium potential and resting membrane potential using a manipulative model. You also know how to calculate equilibrium potential and resting membrane potential using the Nernst and Goldman equations. In addition, you have also drawn graphs to represent the membrane potentials. Activity 7.6 helps you to revise the significance of equilibrium potential.
ACTIVITY 7.6: Significance of equilibrium potential Please draw line graphs showing EK, ENa, ECl, and EM complete with labeled X and Y axis when a cell is at rest. What happens to the membrane potential when permeability to each of the ions increases/decreases? Illustrate by drawing the graphs superimposed on the previous graphs. What may cause the change in permeability of the membrane to certain ions? Hint: what are the specific stimuli?

In Activity 7.7 you will demonstrate the change in membrane potential by working on the template model (use the model in Appendix A in Module 6).
ACTIVITY 7.7: Demonstration on change in membrane potential Set up your membrane template (Appendix A in Module 6) with a K channel in one gap, + and a Na channel in the other (Appendix B in Module 6). To each of these channels lay a gate across the opening on the outside of the membrane. The small notch in the gate should face up. Add the following ions to the ISF: 10 Na , 1 K , and 3 Cl . Add the following ions to the + + cytoplasm: 1 Na , 10 K , and 1 Cl . Please do not disregard the 20 negative charges inside the model cell due to protein. Note that these do not represent physiological proportions, but will be useful for helping you do the simulations. Determine by the counting technique you learned in Module 6 what the membrane potential is for this model cell. Observe the channels that you set up on your template. What types of channels are these? Specify both the ion and the means of opening/closing. One type of gated channel that is vital for communication between cells in the body is controlled chemically. What does this mean?
+ + +

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These chemical-gated (or ligand-gated) channels are found, for example, on the skeletal muscle cells innervated by the motor neurons. So when you decide to flex your arm at the elbow (Fig. 7.1), the signal is sent out to the skeletal muscle cells of the biceps brachii muscle by means of motor neurons that signal the muscle to contract. In this example, the motor neurons release a chemical signal (a neurotransmitter called acetylcholine) that attaches to the channel protein at a special receptor site. The binding of the chemical signal to the receptor causes the gate to open briefly, causing membrane depolarization. Activity 7.8 helps you to understand the above concept better.
ACTIVITY 7.8: Demonstration of depolarisation Continue your work from Activity 7.7. Choose the square signal molecule for your model, + attach it to the receptor site on the gate of the Na channel, and rotate the channel to an open position. Demonstrate what happens to the sodium ions. The attachment between the signal molecule and the receptor site is weak. Soon it detaches and the gate closes. Demonstrate it. The signal molecule is then split apart by an enzyme (not shown here) in the cell membrane. You can simulate this by simply removing the square signal molecule. Based upon what you have learnt so far, predict what will happen to membrane potential + when Na gate is opened. Test your prediction by running the following simulation: Attach the signal molecule to the + channel, open the gate, move 3 Na into the cell, and close the gate. Determine the new + membrane potential and record it. What forces tend to move Na into the cell? If you did it correctly, there is a change in membrane potential called depolarization of the membrane. What does depolarization mean? Draw a graph showing resting membrane + potential and how this potential changes when Na channel is open. Confirm that the potential difference across the membrane was reduced (went closer to zero). Thus depolarization takes the membrane from its resting potential closer to zero. Try to produce a computer animation of the above process.

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In Activity 7.8, the influx of Na+ ions creates a new gradient inside the cell (i.e. charge difference underneath the membrane between the site of Na+ entry and the adjacent sites i.e. the dark band in Fig. 7.6a). To understand this, consider the concentration of sodium ions in the cytoplasm directly beneath the channel compared to the concentration in the adjacent area. What is the gradient? Refer to Fig 7.6a. What do you expect will happen to the extra Na+ ions that just came in? The resulting diffusion of ions creates a tiny current in the cell. It also will restore the local membrane potential back toward the resting value (Fig 7.6b). Fig. 7.7 shows what happens in a graphical format.

Figure 7.6 a. Stimulation of the membrane causes deplarisation; b. The positive charges spread to the neighbouring sites

Figure 7.7. Graphical representation of the spread of charges from the site of depolarization.

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Now you know what happens when the cell is stimulated by a ligand that sits on its receptor, causing the gated ion channel to open. If the stimulus causes Na+ channel to open, Na+ will rush in, depolarizing the membrane from its resting state. The Na+ ions will be attracted to the negative charges of the neighbouring sites and soon the membrane potential falls back to the resting state. This type of transient depolarization is called graded potential. Activity 7.9 helps you to explore more about graded potentials.
ACTIVITY 7.9: Graded potential What is graded potential? Can graded potential be used to carry information? Predict the function of graded potential.

Table 7.1 gives a brief description of different types of membrane potentials.

Table 7.1. Types of membrane potentials Types of potential Potential = potential difference Membrane potential Equilibrium potential Description The voltage difference between two points The voltage difference between the inside and outside of the cell The voltage difference across a membrane that produces a flux of a given ion species that is equal but opposite the flux due to the concentration gradient affecting the same ion species The steady transmembrane potential of a cell that is not producing an electrical signal A potential change of variable amplitude and duration that is conducted decrementally; it has no threshold or refractory period A brief all-or-none depolarisation of the membrane, reversing polarity in neurons; it has a threshold and refractory period and is conducted without decrement A graded potential change produced in the postsynaptic neuron in response to release of a neurotransmitter by a presynaptic terminal; it may be depolarising (EPSP) or hyperpolarising (IPSP) A graded potential produced at the peripheral endings of afferent neurons (or in separate receptor cells) in response to a stimulus A spontaneously occurring graded potential change that occurs in certain specialised cells

Resting potential Graded potential

Action potential

Synaptic potential

Receptor potential

Pacemaker potential

In summary, graded potential occurs only at specific locations on excitable tissues e.g. nerve endings, post-synaptic junctions and pacemaker cells, where ligand-gated or mechanical-gated ion channels exist. Appropriate stimuli stimulate these ion channels, resulting in slow depolarization or hyperpolarisation. This is graded potential.

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2.2. Graphical representation of graded potential Membrane potentials change rapidly over time. In a matter of a few milliseconds (onethousandth of a second) major shifts can occur in membrane potentials with the production and transmission of signals among cells of the body. These changes are often represented graphically as shown in Fig 7.8. This graph represents a recording, over time, of the membrane potential from a single cell. You may remember that the recording electrode is inside the cell (cytoplasm) while the reference electrode is outside in the ISF (Fig 6.4, Module 6).

20 0 Membrane potential (mV) -20 -40 -60 -80 -100 a b c g d h

e f
1 2

6 7

9 10 11 12 13 14 15

Time (msec)
Figure 7.8. Change in membrane potential when a cell is stimulated

The graphs in Fig 7.8 shows a recording of what happens to the membrane potential when 2 separate cells are stimulated (at the arrow, time = 1 msec.) Activity 7.10 helps you to analyse the graph clearly.

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ACTIVITY 7.10: Analysing membrane potential graphically Refer to Fig. 7.8. Look at the top curve abcd and answer the following questions: a. What is the membrane potential at a called? What is its value? b. What is the name of the event when the curve goes up at b? b. Referring to your work in the prior section, what change in the membrane is likely to have caused the shift in membrane potential at b? c. What is the value of membrane potential at the peak of the curve? d. What is the name of the event when the curve goes down at c? e. What change in the membrane is likely to have caused the shift in membrane potential at c? f. Is there any difference between the values of a and d? Why? The lower curve efgh in Fig 7.8 comes from a recording in a different cell. a. What is its resting membrane potential, e? b. What is the name of the event when the curve goes down at f? c. Referring to your work in the prior section, what change in the membrane is likely to have caused the shift in membrane potential at f? d. What is the most negative reading on the lower curve? e. What is the name of the event when the curve goes up at g? f. What change in the membrane is likely to have caused the shift in membrane potential at g? g. Is there any difference between the values of e and g? Why? Fig 7.9 repeats the same concept of depolarization and hyperpolarisation presented in Fig. 7.8. Study the graphs carefully. Where do you think the phenomenon represented by each graph occurs in your body?

Figure 7.9. Membrane potential showing a) depolarization and b) hyperpolarisation

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2.3. Analysis of graded potential In this section you will explore: What is graded potential? Where does it occur? What is it used for? Characteristics of graded potential: Short-lived, local changes in membrane potential (depolarisation or hyperpolarisation) due to opening of chemical-gated ion channels (Fig 7.10a) Amplitude proportional to strength of stimulus (Fig 7.10b) Current flows decreases with distance; so, cannot be utilized for long distance signals (Fig 7.10c) Can be summed spatially and temporally (concept of temporal and spatial summation) (Fig 7.10d)

Activity 7.11 helps you to appreciate the characteristics of graded potential by explaining the molecular mechanisms involved.

Figure 7.10. Characteristics of graded potential

Activity 7.12 helps you to identify the different types of graded potentials that occur in ACTIVITY 7.11: Molecular mechanism of graded potential your body.
For each of the characteristics mentioned above, please explain the molecular mechanism involved.

ACTIVITY 7.12: Types of graded potentials Specific names of graded potential based on location/ function: Receptor potential or generator potential Postsynaptic potential (EPSP and IPSP) Pacemaker potential End plate potential Please provide appropriate diagrams (your own sketch or drawing from a source) to show the location where the above graded potentials occur.

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The graded potential events represented in Fig 7.10 are frequent occurrences in neuron and muscle cells. For example, the brief depolarization of the membrane of a post-synaptic neuron in response to a chemical signal (neurotransmitter) is called an excitatory postsynaptic potential (EPSP). It is so named because it increases the likelihood that the cell will produce an action potential, another type of communication signal (Section 4). The hyperpolarization in Fig 7.10 is called an inhibitory post-synaptic potential (IPSP). These signals have the opposite action of EPSPs. They decrease the likelihood that the cell will produce an action potential. Some neurotransmitters cause depolarization while others cause hyperpolarisation.
ACTIVITY 7.13: EPSP and IPSP

Compare and contrast between EPSP and IPSP. What is the relevance of EPSP and IPSP?

Activity 7.14 helps you to present graded potential in graphical format.

ACTIVITY 7.14: Graphs of graded potential

In Fig 7.11 draw a graph from the data in the table provided. The recordings in the table are from two different cells. Plot the points and connect them to make the curve. When you have completed the graphing, label the segments of each curve by referring to Fig 7.8. (A new segment begins with a change in direction of the curve). You may need to use more letters on the top curve than the bottom. Next, interpret in words what is happening in each segment of the curve. Suggest what is happening in the membrane to bring about the changes from one segment to the next.

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Time (msec) 0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0

Membrane potential (mV)

Membrane potential (mV) Cell 1 Cell 2 -60 -75 -60 -75 -60 -75 -50 -75 +20 -84 -50 -88 -60 -84 -65 -75 -63 -75 -60 -75 -60 -75

20 0 -20 -40 -60 -80 -100

1 2 3 4 5 6 7 8

Time (msec)
Figure 7.11. Membrane potential recordings for two cells

What is the relevance of graded potential? If it cant carry signals over long distance, what is it for? Graded potential is actually important in initiating action potentials, the long distance signals. How is this possible? Before answering the above questions, lets first look at the concept of threshold.

3. Concept of threshold
What is threshold stimulus? It is a stimulus which is strong enough to cause formation of action potential. What is threshold potential? It is a membrane potential (voltage) which is strong enough to stimulate voltage-gated channels to open. It is normally ~15mV above resting potential. To understand the concept of threshold stimulus and threshold potential, lets carry out the Activity 7.15:

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ACTIVITY 7.15: Stimulus-response relationship Study the two panels in Fig. 7.12. Both panels are drawn on the same time scale. The Y-axis of the upper panel represents membrane potential, and that of the lower panel represents stimulus strength. a. Label the Y-axis of the upper panel with -70 mV, -55 mV, 0 mV, and +30 mV. You can disregard the square markings. b. For this simulation, lets use an arbitrary unit for the stimulus strength (which could be solute concentration, strength of mechanical force, etc). Lets assume that the threshold stimulus is 10 units, and the threshold potential is -55 mV (i.e. 15 mV above resting potential). Label the Y-axis of the lower panel with 0 units and 10 units. c. Look at stimulus a. Since its strength is below threshold, it is called a subthreshold stimulus. When you give this stimulus to a cell, its going to open up a few chemical+ gated Na channels, therefore there is a little depolarization (observe the corresponding membrane potential response in the upper panel). This depolarization is below threshold potential, and it dies off when the stimulus is taken off. d. Look at stimulus b. It is still subthreshold, but the strength of the stimulus is greater than a. What do you expect to happen to the cell membrane? The corresponding membrane potential in the upper panel reflects an appropriate depolarization response to stimulus b. e. Stimulus c is greater than b, but is still subthreshold. Draw the appropriate depolarization on the upper panel. f. Stimulus d is greater than c, but still subthreshold. Draw the appropriate depolarization on the upper panel. Based on this activity so far, summarise the characteristics of graded potential. g. Now, look at stimulus e. It has reached threshold, so it is called a threshold stimulus. That means the stimulus is strong enough to open enough ligand-gated channels to + depolarize the membrane to a level where it will stimulate voltage-gated Na channels to open. The characteristic feature of this voltage-gated channel is that all of the channels present at the point of stimulation will open. What do you expect to happen to the + movement of Na ions? This characteristic is also known as all-or-none, i.e. all of the voltage-gated channels either open together under the influence of a threshold stimulus, or none of them will open when the stimulus is subthreshold. Another characteristic feature of the channel is that the opening is transient (only for 1 msec). What do you think will happen when the channels are closed? The spike of depolarization due to simultaneous opening of voltage-gated channels together with repolarisation when the channels are closed is called an action potential. Please refer to the membrane potential (upper panel) in response to stimulus e. h. Stimulus f is greater than threshold stimulus, so it is called a suprathreshold stimulus. + Remember that with threshold stimulus, all the voltage-gated Na channels are open. What do you expect to happen when the cell is given a suprathreshold stimulus? Please draw the membrane potential appropriately as a response to stimulus f. i. Stimuli g and h are also suprathreshold. Please draw the membrane potential appropriately as a response to the stimuli. j. Label subthreshold stimulus, threshold stimulus and suprathresold stimulus, graded potentials, action potentials. k. In addition, another characteristic of a graded potential is it can be summed temporally and spatially. Please refer to Fig. 7.13 and explain the meaning of the terms spatial summation and temporal summation.

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Membrane potential (mV)

Threshold potential Resting membrane potential

Stimulus strength

Threshold stimulus

Time

Figure 7.12. Stimulus-response relationship.

Figure 5.13. Temporal and spatial summation

In summary, action potential can only be produced normally when graded potential has reached threshold. This is the minimum amount of energy required to open the voltagegated Na+ channels that allows depolarisation to take place. The graded potential can be summed spatially or temporally in order to reach threshold.

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4. Change in membrane potentials when stimulated: Action potential

You have seen in the previous section the relationship between graded potential and action potential. Action potential is basically a brief reversal of membrane potential with a total amplitude (change in voltage) of about 100mV (from 70 to +30 mV).
ACTIVITY 7.16: Action potential What does reversal of membrane potential mean? Only cells with excitable membranes (neurons and muscle cells) can generate action potential. Why? Please give a molecular evidence.

Review: Action potentials occur in certain parts of the cell membrane where there are voltage-gated channels. Nerve impulse is transmitted action potentials. It is used as a principal method of communication by neurons Stimulus for generation of action potential is graded potential. It involves voltagegated channels at axon hillocks or proximal to receptor regions An action potential is of no use in terms of signal carrying capacity unless it can be propagated as impulses. In neurons, action potentials trigger neurotransmitter release; in muscle cell they cause contraction. In interpreting the segments of the curve that make up the action potential, note that for a brief period of time, the membrane potential is actually reversed. Locate this portion of the action potential and label it "reversal of membrane potential". The upward section of the curve in Fig. 7.11 between 1.5-2.0 msec is due to the opening of voltage-activated Na+ channels that allow Na+ to flow briefly into the cell. The downward segment from 2.0-2.5 msec is due to outward movement of K+ ions through voltage-activated channels. The period of hyperpolarization between 3.0-4.5 msec results from the fact that more K+ leave the cell than required to bring it back to resting potential. Lets now look closely at the mechanism of action potential production: Formation of action potential involves 3 consecutive but overlapping changes in membrane permeability due to opening and closing of active ion gates: i. Transient increase in Na+ permeability depolarisation ii. Na+ impermeability repolarisation iii. Short-lived increase in K+ permeability

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There are two types of gates on voltage dependent Na+ channels: i. Activation gate ii. Inactivation gate Please use Fig. 7.14 to figure out the involvement of the gates in the formation of an action potential.

Figure 7.14. Mechanism of action potential formation.

ACTIVITY 7.17: Mechanism of action potential production Study Fig 7.14 carefully and write a short assay on the mechanism of action potential production. Fig 7.15 will help you to explain the ionic mechanism associated with the phases (1-4) of action potential.

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Fig. 7.15. Ionic mechanism associated with the phases of action potential

The depolarization phase of the action potential generated when the membrane reaches threshold actually has a positive-feedback relationship (Fig. 7.16). The threshold potential causes opening of the voltage-gated Na+ channels which will increase the permeability of the membrane to Na+. This will increase the flow of Na+ into the cell and will enhance depolarization. The positive feedback continues until all the voltage-gated Na+ channels are open. When the voltage-gated Na+ channels close, the membrane repolarises (Fig. 7.16).

Fig. 7.16. Positive-feedback relationship during depolarization phase of action potential

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You should be comfortable with the relationship between graded potential and action potential now. Table 7.2 describes the comparison between graded potential and action potential.
Table 7.2. Comparison between graded potential and action potential Action potential All-or-none once membrane is depolarised to threshold. Amplitude is independent of initiating event Cannot be summed Threshold is usually about 15 mV depolarised relative to the resting potential Has refractory period Conducted without decrement Duration constant Depolarisation with overshoot Initiated by graded potential Mechanism depends on voltage-gated channel

Graded potential Amplitude varies with conditions of the initiating event Can be summed Has no threshold Has no refractory period Conducted decrementally Duration varies with initiating condition Can be depolarised or hyperpolarised Initiated by environmental stimulus, neurotransmitter or spontaneously Mechanism depends on chemical/mechanical-sensitive channel

You know that action potential is of no use in terms of signal carrying capacity unless it can be propagated. Let us now explore how action potential is propagated and form what is known as impulses.

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5. Conduction/propagation/transmission of action potential

5.1. Mechanism of action potential propagation An action potential is of no use if it cannot be propagated along the entire length of the neuron or muscle fibre. Why? How is action potential propagated? The first action potential in a neuron results from a graded potential (receptor potential, synaptic potential, end-plate potential, or pacemaker potential) that is still at threshold when it reaches the voltage-gated channel at the axon hillock. If this action potential is not propagated, the signal that establishes the graded potential will not be transmitted. To appreciate the above concept, lets do Activity 7.18.
ACTIVITY 7.18: Mechanism of action potential propagation Fig. 7.17. Formation of consecutive action potential. Note the of chemical-gated, Review the positions timing of action potential + formation in the three panels. mechanically-gated and voltage-gated Na channels on the membrane of a neuron. Follow the explanation below while referring to Fig. 7.17.

Graded potential (formed as receptor potential, synaptic potential, end-plate potential, or pacemaker potential) + stimulates voltage-gated Na channel formation of the first action potential + (depolarisation i.e. Na high inside): Top panel of Fig. 7.17 Membrane depolarization of the first action potential stimulates the adjacent voltage-dependent channels to open, thus a new action potential is produced (Middle panel in Fig. 7.17). The second action potential stimulates the third, and so on (bottom panel in Fig. 7.17). As long as the stimulus is there to produce the initial action potential, it will continuously be propagated along the axon of the neuron This is like a domino effect, which you will work on in Activity 7.19.

Figure 7.17. Mechanism of action potential propagation

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Activity 7.19 helps you to feel the process of action potential propagation and to relate it to the strength of the stimulus.
ACTIVITY 7.19: The domino effect of action potential propagation Arrange about 20 dominos on the floor.

First give a little push to the first domino; use a force that is not sufficient to cause the domino to fall. What do you notice? Relate it to the subthreshold stimulus. The stimulus you gave was not strong enough to cause the first domino to fall onto the second domino, just like a subthrehold stimulus is not strong enough to depolarize the neuron to open the voltage-gated channel. Give a stronger push onto the first domino until it is just sufficient to fall onto the second domino. What do you notice? Once initiated, the action potential is a self propagating process that continues along the axon at a constant velocity (like domino effect). Relate this to the events that take place in a neuron when a threshold stimulus is applied. Hypothesise what would happen if you push the first domino harder. Verify this by timing the fall of the 20 dominos. Compare this to the threshold and suprathreshold effects of stimuli. Please use the analogy of the connection between one domino and the next domino to the connection between one action potential to the next action potential in a neuron. Please explain based on the molecular level.

In summary, transmission of action potential automatically occurs once an action potential is established as a result of receptor stimulation. This is due to the presence of voltage gate ion channels present in the membrane of excitable cells.

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5.2. The significance of refractory periods Why is action potential always propagated in a one-way direction i.e. away from the point of origin towards the axon terminal? The answer is straightforward; it is because naturally the neuron is always stimulated at the dendrite or soma region and therefore voltage-gated Na+ channels immediately distal to it (e.g. at the axon hillock) open consecutively from the point of stimulation towards the axon terminal. Moreover, during the depolarizing phase of the action potential, the positive charges cannot cause depolarization of the preceding site because the site is in refractory period. This is the reason why propagation of action potential is unidirectional. What is refractory period? Absolute refractory period: a period when no action potential can be formed regardless of the strength of the stimulus Relative refractory period: a period when action potential can only be formed when the stimulus is stronger than the normal threshold. Activity 7.17 helps you to understand more about the relevance of refractory periods.

Fig. 7.18. Absolute and relative refractory periods

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ACTIVITY 7.20: Relevance of refractory periods Refer to Fig. 7.18 (this is the same figure as 7.15). What is: Absolute refractory period? Relative refractory period? Please explain the molecular mechanism that causes absolute refractory period and relative refractory period. Hint: you must know what happens to the channel proteins during depolarization, repolarisation, and after-hyperpolarisation (undershoot). Why cant depolarization of the membrane immediately distal to the patch that is undergoing refractory period produce action potential on that patch?

In summary, refractory period is an important membrane potential phenomenon that allows unidirectional flow of action potential. This ensures that information is transmitted in one direction only. In other words, this prevents crossing of information throughout the nervous system which would cause a lot of communication problems.

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5.3. Strategies to hasten action potential propagation

Signals of some information need to be propagated very quickly, while others do not require very fast delivery. For example, signals from peripheral touch receptors are propagated quickly to the brain, whilst signals via the autonomic nervous system need not be sent as quickly. What are the strategies that can hasten the rate of propagation of action potentials? a. the presence of myelin sheath b. diameter of the axon: larger diameter permits faster conduction c. influence of chemicals Myelin sheaths on axons have significant effect on the rate of action potential conduction (Fig. 7.19). Voltage gated channels are mostly at the nodes of Ranvier. The myelinated portion of the axon does not permit much ion exchange due to the sheaths insulating property. Therefore, current carried by Na+ and K+ flows through the membrane mainly at the nodes. In other words, action potentials occur only at the nodes. This hastens the rate of propagation of action potentials. This is called saltatory conduction. Saltatory conduction is analogous to walking with normal steps, whereas continuous conduction is like walking by arranging one heel to touch the toes of the other leg.

Fig. 7.19. Mechanism of salutatory conduction of action potentials

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ACTIVITY 7.21. Saltatory conduction of action potential Based on Fig. 7.19 and 7.20: explain the molecular mechanism of saltatory conduction compare and contrast between continuous and saltatory conduction. explain the significance of saltatory conduction. which neurons in the body are myelinated?

Axon diameter also affects the rate of action potential conduction. Larger diameter axons propagate impulses faster than smaller ones. Therefore, information that requires vey fast response (for example sensory inputs and motor outputs to muscles and joints) is transmitted by thicker fibres. Effect of axon diameter on speed of conduction: Group A fibres: largest diameter ~5-20 mm; all myelinated; somatic sensory (touch, pressure) and motor fibres to skin, muscle and joints; speed of conduction: 12-130 m/sec Group B fibres: diameter 2-3 mm; lightly myelinated; conduct sensory impulses to CNS, and all ANS axons to autonomic ganglia; speed: up to 15 m/sec Group C fibres: diameter 0.5-1.5 mm; all unmyelinated; conduct sensory impulses for 34

pain, touch, pressure, heat and cold from the skin, and pain impulses from the viscera. Include autonomic fibres from autonomic ganglia to heart, smooth muscle and glands. Speed: 0.5-2m/sec) Chemical factors can also affect impulse conduction. These are substances which alter membrane permeability to ions, thus influence the rate of impulse conduction. For example: Ca2+ are required to close Na+ channels in axon membranes during an action potential. Lack of Ca2+ Na+ channels remain open Na+ diffuse in again and again impulses transmitted repeatedly. If impulses to skeletal muscle muscle spasm (tetanus). Usually occur in women during pregnancy, people with diet lacking Ca2+ or Vit D, or prolong diarrhea. A small increase in concentration of extracellular K+ resting potential of nerve fibres less negative threshold potential reached with smaller stimulus i.e. the affected fibers are very excitable convulsion may occur. A big decrease in concentration of extracellular K+ resting potential very negative action potential cannot occur muscles become paralysed. Certain anaesthetic drugs e.g. procaine decrease membrane permeability to Na+ prevent impulses from passing through the affected area prevent perception of touch and pain. In summary, structural modification of the axons helps in determining the velocity of action potential transmission. Axons that have to transmit action potential fast are bigger in size and are myelinated. In addition, chemicals in the extracellular fluid around the axons would also affect transmission of action potentials.

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vii. Summary
The resting membrane potential itself cannot propagate impulses, thus it cannot transmit information signals. However, it is a basis for production of graded potentials when the membrane is stimulated. This is due to the membrane characteristics of excitable cells i.e. by having gated ion channels that are responsive to chemicals, change in voltage, or mechanical stimulation. The action potentials produced are propagated along the axon due to the presence of voltage-gated ion channels on the axon membrane. This is how information signal is transmitted from the receptor (sensor) to the central nervous system (CNS) for processing, and from the CNS to the effector for appropriate response to the stimulus.

ACTIVITY 7.22: Summary Draw a creative and comprehensive concept map that encompasses the main ideas in this module. Describe what you have learnt from this module, including non-academic outcomes. Comments on the activities, and suggest innovations for improvement of the module.

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viii. Conclusions
Please make sure that you achieve all the objectives set up at the beginning of this module: Objectives 1. Discuss the role of the nervous system in intercellular communication. 2. Discuss the sequence of events that take place on a cell membrane when a stimulus reaches a receptor, resulting in a change in membrane permeability to certain ions, consequently changing the membrane potential. 3. Describe the significance of the presence of different ion channels in the membrane of a neuron in terms of their effects on membrane potential. 4. Define graded potential, describe how it is established, and describe its characteristics. List down the types of graded potential (including EPSP and IPSP) and state the significance of each. 5. Plot the membrane potential response to subthreshold, threshold, and suprathreshold stimuli and explain the molecular basis of the response. Discuss their significance. 6. Explain the meaning of action potential; describe the mechanism of action potential production and action potential propagation. 7. Compare and contrast between action potential and graded potential. 8. Describe the molecular basis of absolute and relative refractory periods and state the significance. 9. Describe the strategies that can hasten action potential propagation. Comments

Dont forget the objectives that you have constructed yourselves!

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ix. Assessment
1. Match each phase of an action in the following list with the appropriate description below: depolarization (D), repolarization (R), after-hyperpolarization (AH). Some statements may have more than one right answer. a. ______ fast, inward movement of sodium b. ______ membrane potential goes from 70 mV to 80 mV c. ______ fast, outward movement of potassium d. ______ membrane potential goes from +30 mV to 70 mV e. ______ activation and inactivation gates of Na+ channels are both open f. ______ membrane potential goes from 70 mV to +30 mV g. ______ potassium gates are open

2. Fill in the blanks: Action potentials are rapid depolarizations of the plasma membrane that travel from the (a) to the (b). The action potential is divided into three phases. Depolarization is caused by the opening of (c) channels, which allows this ion to (d) the cell. The repolarization phase is caused by the closing of (e) channels and the opening of (f ) channels; this allows potassium to (g) the cell. After hyperpolarization occurs because (h) channels are slow in closing, allowing continued movement of potassium (i) of the cell for a brief period of time. a. _________________________ f. _________________________ b. _________________________ g. _________________________ c. _________________________ h. _________________________ d. _________________________ i. _________________________ e. _________________________

3. Label the following statements as true (T) or false (F). If false, change the statement to make it true. ______ During the depolarization phase of an action potential, the membrane potential changes from approximately +30 mV to 70 mV. ______ Saltatory conduction is seen in unmyelinated axons. ______ Myelinated axons conduct nerve impulses faster than unmyelinated axons. ______ The membrane potential during after-hyperpolarization goes from approximately 70 mV to +30 mV. ______ A subthreshold stimulus at the axon hillock initiates an action potential.

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______ If given enough of a stimulus, an action potential can be stimulated during the relative refractory period. ______ Action potentials tend to be graded in their response. ______ The positive feedback loop of the opening of sodium activation gates is initiated at threshold. ______ During repolarization, voltage-gated potassium gates are open.

4. Place the following events of an action potential in the correct order. a. Sodium permeability decreases rapidly and potassium permeability increases. b. Potassium exits the cell rapidly. c. After-hyperpolarization occurs. d. Potential returns from approximately +30 mV back to 70 mV. e. Potassium permeability remains elevated briefly after reaching 70 mV. f. Membrane potential changes from approximately 70 mV to +30 mV. g. rapid sodium influx into the cell h. an increase in sodium permeability Sequence: ____________________________

5. Assume that a nerve cell has just been stimulated at a dendrite and an action potential will take place in that neuron. Place in sequence the following locations with their associated potentials. a. axon hillock : action potential b. axon terminal : induces a synaptic potential in postsynaptic cell c. axon : action potential d. dendrite : graded potential Sequence: ____________________________

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