LIU Chuan Yong
刘传勇
Institute of Physiology
Medical School of SDU
Tel 88381175 (lab)
88382098 (office)
Email: liucy@sdu.edu.cn
Website: www.physiology.sdu.edu.cn
1
Section 2
Bioelectrical Phenomena
of the Cell
2
Basic Concepts
Volt
A charge difference between two points in
space
3
Basic Concepts
Ions – charged particles
Anions – Negatively charged particles
Cations – Positively charged particles
4
Basic Concepts
Forces that determine ionic movement
Electrostatic forces
Opposite charges attract
Identical charges repel
Concentration forces
Diffusion – movement of ions through
semipermeable membrane
Osmosis – movement of water from region of
high concentration to low
5
Selective Permeability of
Membranes
Some ions permitted to cross more easily than
others
Neuronal membranes contain ion channels
Protein tubes that span the membrane
Some stay open all the time (nongated)
Some open on the occasion of an action potential,
causing a change in the permeability of the
membrane (gated)
6
I. Membrane Resting Potential
A constant potential difference across the resting cell
membrane
Cell’s ability to fire an action potential is due to the
cell’s ability to maintain the cellular resting potential
at approximately –70 mV (-.07 volt)
The basic signaling properties of neurons are
determined by changes in the resting potential
7
Membrane Resting Potential
 Every neuron has a separation of electrical charge
across its cell membrane.
 The membrane potential results from a separation of
positive and negative charges across the cell membrane.
8
Membrane Resting
Potential
 excess of positive charges
outside and negative charges
inside the membrane
 maintained because the lipid
bilayer acts as a barrier to
 gives rise to an electrical
potential difference, which
ranges from about 60 to 70
mV.
 (Microelectrode) 9
Concept of Resting Potential (RP)
 A potential difference across the cell
membrane at the rest stage or when the
cell is not stimulated.
 Property:
 It is constant or stable
 It is negative inside relative to the outside
 Resting potentials are different in different
cells.
10
Ion Channels
Two Types of Ion Channels
Gated
Non-Gated
11
Resting Membrane Potential
Na+ and Cl- are more concentrated outside the
cell
K+ and organic anions (organic acids and
proteins) are more concentrated inside.
12
Intracellular vs extracellular ion concentrations
Ion
Intracellular
Extracellular
Na+
K+
Mg2+
Ca2+
H+
5-15 mM
140 mM
0.5 mM
10-7 mM
10-7.2 M (pH 7.2)
145 mM
5 mM
1-2 mM
1-2 mM
10-7.4 M (pH 7.4)
Cl-
5-15 mM
110 mM
13
Resting Membrane Potential
 Potassium ions, concentrated inside the cell tend to move
outward down their concentration gradient through
nongated potassium channels
 But the relative excess of negative charge inside the
membrane tend to push potassium ions out of the cell
14
Potassium equilibrium
-90 mV
15
Resting Membrane
Potential
Na+ is more concentrated
outside than inside and
therefore tends to flow into the
cell down its concentration
gradient
Na+ is driven into the cell
by the electrical potential
difference across the
membrane.
• But what about sodium?
• Electrostatic and Chemical forces act together on
Na+ ions to drive them into the cell
• The Na+ channel close during the resting state
16
+
Na
electrochemical gradient
17
Equilibrium Potentials
Theoretical voltage
produced across the
membrane if only 1 ion
could diffuse through the
membrane.
If membrane only
permeable to K+, K+
diffuses until [K+] is at
equilibrium.
Force of electrical
attraction and diffusion
are = opposite.
18
Calculating equilibrium potential
Nernst Equation
Allows theoretical membrane potential to be
calculated for particular ion.
Membrane potential that would exactly balance
the diffusion gradient and prevent the net
movement of a particular ion.
Value depends on the ratio of [ion] on the 2 sides
of the membrane.
19
Nernst equation
Equilibrium potential (mV) , Eion
[C]o
RT
=
ln
zF
[C]i
where,
[C]o and [C]i = extra and intracellular [ion]
R = Universal gas constant (8.3 joules.K-1.mol-1)
T = Absolute temperature (°K)
F = Faraday constant (96,500 coulombs.mol-1)
z = Charge of ion (Na+ = +1, Ca2+ = +2, Cl- = -1)
For K+, with [K+]o = 4 mmol.l-1 and [K+]i = 150 mmol.l-1
At 37°C, EK = -97mV ENa = +60mv
20
+10
Experimental points
Membrane
potential
(millivolts)
-60
-70
(Red line shows values
according to Nernst equation)
-130
1
5
10
100
Extracellular potassium concentration (millimoles)
[K+]o = 4 mmol.l-1
21
Resting Membrane Potential
Resting membrane potential is less than Ek because
some Na+ can also enter the cell.
The slow rate of Na+ influx is accompanied by slow
rate of K+ outflux.
Depends upon 2 factors:
Ratio of the concentrations of each ion on the 2
sides of the plasma membrane.
Specific permeability of membrane to each
different ion.
Resting membrane potential of most cells ranges
from - 65 to – 85 mV.
22
The Sodium-Potassium Pump
extrudes Na+
from the cell
while taking
in K
• Dissipation of ionic gradients is ultimately
prevented by Na+-K+ pumps
23
Resting Potential
24
The formation of resting potential
depends on:
 Concentration difference of K+
across the membrane
 Permeability of Na+ and K+ during
the resting state
 Na+-K+ pump
25
Factors that affect resting potential
 Difference of K+ ion concentration
across the membrane
 Permeability of the membrane to Na+
and K+.
 Action of Na+ pump
26
Basic Electrophysiological Terms I:
 Polarization: a state in which membrane is
polarized at rest, negative inside and positive
outside.
 Depolarization: the membrane potential
becomes less negative than the resting potential
(close to zero).
 Hyperpolarization: the membrane potential is
more negative than the resting level.
27
Basic Electrophysiological Terms I:
 Reverspolarization: a reversal of
membrane potential polarity.
 The inside of a cell becomes positive
relative to the outside.
 Repolarization: restoration of normal
polarization state of membrane.
 a process in which the membrane potential
returns toward from depolarized level to the
normal resting membrane value.
28
II Action Potential
Successive Stages:
(2)
(1) Resting Stage
(3)
(2) Depolarization stage
(1)
(4)
(3) Repolarization stage
(4) After-potential stage
29
Concept
 Action potential is a rapid, reversible, and
conductive change of the membrane
potential after the cell is stimulated.
 Nerve signals are transmitted by action
potentials.
30
Action Potential
Sequence
• Voltage-gated Na+ Channels open and Na+
rushes into the cell
31
Action Potential
Sequence
•
At about +30 mV, Sodium channels close, but now,
voltage-gated potassium channels open, causing an
outflow of potassium, down its electrochemical gradient
32
Action Potential
Sequence
equilibrium potential of the cell is restored
33
Action Potential
Sequence
• The Sodium – Potassium Pump is left to clean
up the mess…
34
Ion Permeability during the AP
35periods
Figure 8-12: Refractory
Basic Electrophysiological Terms II (1)
 Excitability: The ability of the cell to generate
the action potential
 Excitable cells: Cells that generate action
potential during excitation.
 in excitable cells (muscle, nerve, secretery cells), the
action potential is the marker of excitation.
 Some scholars even suggest that in excitable
cells, action potential is identical to the
excitation.
36
Basic Electrophysiological Terms II (2)
Stimulus: a sudden change of the (internal or
external) environmental condition of the cell.
includes physical and chemical stimulus.
The electrical stimulus is often used for the
physiological research.
Threshold (intensity): the lowest or minimal
intensity of stimulus to elicit an action
potential
(Three factors of the stimulation: intensity,
duration, rate of intensity change)
37
Basic Electrophysiological Terms II (3)
 Types of stimulus:
Threshold stimulus: The stimulus with the
intensity equal to threshold
Subthreshold stimulus: The stimulus with
the intensity weaker than the threshold
Suprathreshold stimulus: The stimulus with
the intensity greater than the threshold.
38
Action Potential Summary
 Reduction in membrane potential
(depolarization) to "threshold" level leads to
opening of Na+ channels, allowing Na+ to
enter the cell
 Interior becomes positive
 The Na+ channels then close automatically
followed by a period of inactivation.
 K+ channels open, K+ leaves the cell and the
interior again becomes negative.
 Process lasts about 1/1000th of a second.
39
Properties of the Action Potential
 “All or none” phenomenon
 A threshold or suprathreshold stimulus
applied to a single nerve fiber always initiate
the same action potential with constant
amplitude, time course and propagation
velocity.
 Propagation
 Transmitted in both direction in a nerve
fiber
40
III Initiation of Action
Potential
41
Squid giant axon
42
Gated channel states
43
Na+ Channel a1-Subunit Structure
I
II
III
IV
NH2
Outside
+
+
+
+
+
+
+
+
+
+
+
+
b1
Inside
CO2H
I
F
M
NH2
CO2H
+ + + + + + + +
RVIRLARIGRILRLIKGAKGIR
I
F
M
- Inactivation “Gate”
IVS4 Voltage Sensor
44
Voltage gated
But “ready”
Not “ready”
45
Activation & Fast Inactivation
46
Sodium Activation and Inactivation Variable
vs Voltage
Activation Gate
Inactivation gate
If resting potential depolarized by 15 – 20 mV, then activation
gate opened with 5000x increase in Na+ permeability followed
by inactivation gate close 1 ms later
47
Stimulation
Positive feedback loop
Reach
“threshold”?
If YES, then...
48
Action potential initiation
S.I.Z.
49
Action potential termination
50
51
Threshold Potential
 Threshold potential plays a key role in the genesis of
action potential.
 Threshold potential is a critical membrane potential
level at which an action potential can occur.
 Why can all the Na+ channel open at the threshold
potential? It is dependent on the gating property of the
voltage-gated Na+ channels.
 The value of threshold potential of most excitable cell
membrane is about 15 to 20 mV less negative than the
resting potential.
 The threshold stimulus is just strong enough to
depolarize the membrane to the threshold potential level,
therefore it can cause an action potential.
52
Electrophysiological Method
to Record Membrane
Potential I
Voltage Clamp
53
Andrew Fielding Huxley
“for their discoveries concerning the ionic
mechanisms involved in excitation and
inhibition in the peripheral and central
portions of the nerve cell membrane”
The Nobel Prize in Physiology or Medicine (1963)
Alan Lloyd Hodgkin
54
The voltage clamp
 Cole and colleagues developed a method for maintaining Vm at any
desired voltage level (FBA, Feedback Amplifier)
 Required monitoring voltage changes, feeding it through an amplifier
to drive current into or out of the cell to dynamically maintain the
55
voltage while recording the current required to do so
The Hodgkin-Huxley Model of Action Potential
Generation
56
57
Triphasic response
58
Evidence for a
Sodium Current
Remove extracellular
sodium
59
Modern
proof of
nature of
currents
Use ion
selective
agents
60
 Removing Na+ from the bathing medium, INa becomes
negligible so IK can be measured directly.
 Subtracting this current from the total current yielded
61
INa.
62
Conductance of Na+ and K+ channels
63
Voltage-Dependence of Conductance
64

gNa increases quickly, but
then inactivation kicks in and
it decreases again.
An action potential
 gK increases more slowly,
and only decreases once the
voltage has decreased.
 The Na+ current is
autocatalytic. An increase in
V increases gNa , which
increases the Na+ current,
and increases V, etc.
 Hence, the threshold for
action potential initiation is
where the inward Na+
current exactly balances
the outward K+ current.
65
Bert Sakmann
"for their discoveries concerning the function
of single ion channels in cells"
The Nobel Prize in Physiology or Medicine (1991)
Erwin Neher
66
Patch clamp recording
Suction
"Giga-seal"
1 µm
Cytoplasm
Glass
microelectrode
Ion channels
Cell Membrane
67
68
Single channel record
Closed
4 pA
Open
100 ms
69
One result from patch clamp
studies was the finding that
ion channels conduct currents
in an all or nothing fashion
70
Voltage-dependent Channel
Conductance
71
How channel conductances accumulate
Next page shows an idealized version
72
73
Inactivating Na+ channel currents
74
IV Local Response
75
Graded
(local)
potential
changes
2 x more chemical=
2 x more potential
change
76
Local Response
 Definition:
 Local response is a small change in
membrane potential caused by a
subthreshold stimulus
 Properties:
 It s a graded potential
 Its propagation is electronic conduction
 It can be summed by two ways
 Spatial summation
 Temporal summation
77
Membrane Potential (mV)
Excitatory
a
Excitatory
b
Inhibitory
c
d
a
b
c
d
Spatial
Summation
Time
Spatial
Summation
78
Membrane Potential (mV)
Excitatory
a
Excitatory
b
Inhibitory
c
d
a
b
c
d
Temporal
Summation
Time
Temporal & Spatial
Summation
79
Distribution of channels
Axon Hillock
(Trigger Zone)
Leak channels everywhere
80
Role of the Local Potential
 Facilitate the cell.
 This means it increase excitability of
the stimulated cell
 Cause the cell to excite once it is
summed to reach the threshold
potential
81
82
V. Propagation of the Action Potential
83
84
85
Myelinated neuron of the central nervous system
86
Saltatory conduction:
The action potential jumps from node to node
87
88
Saltatory Conduction
89
Saltatory Conduction
 The pattern of conduction in the
myelinated nerve fiber from node to
node
 It is of value for two reasons:
 very fast
 conserves energy.
90
91
Factors that affect the propagation
 Bioelectric properties of the membrane
 Velocity and amplitude of membrane
depolarization
92
V Excitation and
Excitability of the Tissue
93
Excitation and Excitability of the
Tissue
Review: Excitation and Excitable Cell
Review: Threshold Stimulation and
Excitability
Change of Excitability after the
Excitation
94
95
Slide 3 of 28
96
4. Factors that Determine the
Excitability
 Resting potential
 Threshold potential
 Concentration of extracellular Ca2+
97
Activation Gate
Inactivation gate
m
h
n
If resting potential depolarized by 15 – 20 mV, then activation
gate opened with 5000x increase in Na+ permeability followed
by inactivation gate close 1 ms later
98
•the threshold for action potential initiation is where
the inward Na+ current exactly balances the
outward K+ current.
99
Concentration of extracellular Ca2+
100