3. How do brain cells communicate?

Neuronal communication

Mechanisms of information transfer between different cellular

compartments: intracellular communication

Mechanisms of information transfer between different neurons:

intercellular communication
 3. How do brain cells communicate?

Mechanisms of intracellular
neuronal communication

-- Specific action potential waveforms and specific

patterns of action potential firing in different types of
neurons and their mechanism of generation

--Electrophysiological methods:
experimental protocols to measure the biophysical
properties of voltage-gated ion channels and to
determine the specific ion channels underlying the
different firing properties of different neurons
 There are different types of neurons, with different
molecular, morphological and functional properties,
including different firing properties.

Each type of neuron has a unique set of voltage-gated ion

channels, which determines the specific electrical activity (firing
properties) of the neuron and, hence, its specific role in
information processing in a given neural circuit.

In particular the specific set of voltage-gated ion channels

located in the initial segment of the axon determines:

1. The shape and amplitude of the action potential.

2. The temporal sequence of action potentials in response to a
given synaptic input.
3. The spontaneous activity in certain neurons
 3. How do brain cells communicate?

Mechanisms of intracellular
neuronal communication

-- Specific action potential waveforms and specific

patterns of action potential firing in different types of
neurons and their mechanism of generation

--Electrophysiological methods:
experimental protocols to measure the biophysical
properties of voltage-gated ion channels and to
determine the specific ion channels underlying the
different firing properties of different neurons
 Channels Electrical signals (V change)
biophysical properties properties

Permeation : Erev (and g) Sign (and amplitude) of V change

Gating: Po (V, t) Amplitude of V change

Kinetics of V change

Passive membrane properties: C, R Amplitude and kinetics of V change

V = IR;
Membrane time constant = RC

Po (V, t)
G(V, t) = N Po(V, t) g
I (V, t) = G(V, t)(V-E)
I (V, t) = ∑iGi(V, t)(V-Ei) + Ileak
Po= single channel open probability =
fraction of open channels in population of
N channels.
g= single channel conductance
I= macroscopic current carried by
population N of channels
G= macroscopic membrane conductance
Action potential
 VOLTAGE-CLAMP technique

Cole, 1947; Hodgkin and Huxley, 1948

Neher and Sakmann, 1981 Patch Clamp Configurations
1. Antivibration table
2. Faraday cage
3. Microscope
6. Micromanipulator
7-8 joystick and micromanipulator
controller
11 videocamera
13 photomultiplier

Not shown: patch-clamp amplifier,

interface, computer
 1. Identification of the different types of ion channels
contributing to the total cellular current

Membrane
potential

Current
Squid giant axon

Which type of voltage-gated ionic channels mediate the fast

transient inward current and the delayed outward current?
 Erev of Iearly is +52 mV, similar to Nernst potential for
T=3.8 °C
Na+ ions most likely Iearly is a Na+ current (carried
Hodgkin et al., 1952 by channels selective for Na+ ions)
Erev of Ilate is < -50 mV, but the exact value cannot be
inferred from the I-V
 Ion substitution experiments to confirm the selectivity of
the involved ion channels
Iearly is a Na+ current
Na+ ions in the extracellular solution are
substituted with impermeant choline+ ions:
fast transient inward current (Iearly)
becomes outward, thus confirming that
it is carried by Na+ ions;
Ilate is unaltered: it is not carried even
in part by Na+ ions.
Other experiments, including substitution
experiments, have shown that it is a K+
current
 Separation-isolation of the different ionic
components of the total current

1. Isolation via ionic substitution

2. Pharmacological separation
Pharmacological isolation of
different ionic currents

Tetrodotoxin (TTX): specific

inhibitor of voltage-gated
Na+ channels (NaV) Current measured
in the presence of TTX
Tetraethilammonium (TEA):
specific inhibitor of voltage-
gated K+ channels (KV) Current measured
in the presence of TEA
 Isolation of a given ionic current via ionic substitution

The Na+ current component ENa with [Na]ext reduced

is eliminated at different V to 10% of the control
value in physiological
by reducing the Na+ solution (100% Na)
concentration to the value at
V
which ENa=V.
The isolated INa is obtained
as the difference between
the total current and the
remaining current

Hodgkin and Huxley, 1952 T=8.5 °C

 The dependence INa and IK on V reflects the V-dependence of both the
fraction of open channels Po (and hence G) and the single channel current i

IK

INa

T=3.8 °C Hodgkin et al., 1952

 V (mV)

i = g (V-E)
i

Po = 1/ (1+ exp (-(V-V1/2)zF/RT ) )

po
po varies from 0 for V--> -∞ to 1 for V-->+∞

V (mV)

V (mV)

I = NiPo
ipo
I = N Po(V) g (V-E) = G(V) (V-E)
Experimental protocols to derive from
measurements of the whole-cell current I:

the V-dependence of the single channel current i

(and hence Erev)

the V-dependence of G (and hence Po(V)).

 Instantaneous I-V curve
Itail (I2) vs V at constant G (G(V1)
I2 is measured in the first microseconds after repolarization
at different values of V2

The instantaneous I-V is nearly

linear
It gives the V-dependence of the
ion flux through the open channels
(i x No)

I = G(V1) (V-E) (I = Itail =I2)

T= 4°C
I = G(V) (V-E) (I = Ipeak =I1)

G(V) = I / (V-E)

T=20 °C Hodgkin and Huxley, 1952

 Ipeak = N Po(V) g (V-E) = G(V) (V-E)
GNa(V) = NNa gNa Po,Na(V) = INa(V) / (V-ENa)

GK(V) = NK gK Po,K(V) = IK(V) / (V-EK)

GK GNa

G(V) for a given ionic component reflects the voltage dependence

of the fraction of open channels Po(V) (Po(V) = G(V)/Ng)
Po vs V curve is the channel steady-state activation curve
 Experimental protocols to derive the biophysical properties of a given
component of the macroscopic ionic current

I-V (Ipeak vs Vt) if Erev > activation threshold

Instantaneous I-V (Itail vs Vrep at constant Vt) if Erev < activation threshold

 Erev

I-V (Ipeak vs Vt);

Itail vs Vt
 G(V)

 Steady state activation curve: Po (V)

G(V)