April 16, 2010 16:28 WSPC/S0219-6352 179-JIN 00236

Journal of Integrative Neuroscience, Vol. 9, No. 1 (2010) 49–64

c Imperial College Press
DOI: 10.1142/S0219635210002366

Research Report

A METHOD FOR ESTIMATING GAP-JUNCTIONAL

CONDUCTANCE BETWEEN CA3 HIPPOCAMPAL
PYRAMIDAL NEURONS

R. R. POZNANSKI∗, L. S. NG and C. S. WOO

Department of Artificial Intelligence
Faculty of Computer Science and Information Technology
University of Malaya, 50603 Kuala Lumpur, Malaysia
∗
roman.poznanski@um.edu.my

Received 23 December 2009

Accepted 18 February 2010

Coupling ratio analysis using dual intracellular recording combined with ultrastructural
examination of particles in plaques corresponding to gap-junctional channels is a quantita-
tive measure of electrotonic coupling. The difficulty in combining both electrophysiological
and morphological methods in determining the gap-junctional conductance of electrical
synapses is addressed through the use of a passive cable model of two electrotonically
coupled CA3 hippocampal pyramidal cells. Cable estimates of the gap-junctional coupling
conductance based on electrophysiological data are combined with freeze-fracture replicas
of connexin channels (gap-junctions) to confirm strong coupling between the somata of
CA3 pyramidal neurons, based on the assumption that the electrical synapse works as an
ohmic resistor influenced by an ensemble of unitary resistances of gap-junctional channels.

Keywords: Gap-junctions; CA3 hippocampal pyramidal cells; somatic coupling resistance;

electrotonic synapse; cable model; connexin Cx36.

1. Introduction
Dye-coupling between CA3 pyramidal neurons was observed at soma-somatic and
dendro-somatic sites which led MacVicar and Dudek [21] to hypothesize the pres-
ence of gap-junctions on somata and proximal apical dendrites of CA3 pyramidal
cells. Electrotonic coupling is mediated by gap-junctions found in neurons through-
out the hippocampus (see [34] for review) as well as other regions of the mammalian
brain (see [37] for review). In particular, gap-junctions have been observed phys-
iologically and ultrastructurally on CA3 hippocampal pyramidal neurons between
their somatas and dendrites [23, 35], and axons [13, 33]. In the mammalian brain
approximately 12% of all CA3 pyramidal neurons in the hippocampus are electro-
tonically coupled [22]. However, neither the number of electrotonic junctions, nor

∗ Corresponding author.

49
April 16, 2010 16:28 WSPC/S0219-6352 179-JIN 00236

50 Poznanski et al.

Fig. 1. A schematic illustration of a pair of tapered equivalent cables representing the dendrites
each attached to a soma (Z = 0) represented by a RC-circuit and coupled together through a
somatic coupling resistance (Rj ). It is assumed that the gap-junction is non-rectifying with individ-
ual channels acting as passive ohmic conductors.

the strength of the coupling at electrical synapses in the hippocampus is known with
certainty.
Poznanski et al. [30] developed a passive cable model of a pair of neurons cou-
pled via a junctional resistance between cell bodies with the dendrites represented as
tapered equivalent cables (see Fig. 1). In that paper, cable estimates of the somatic
coupling conductance were found based only on electrophysiological data which
resulted in weak coupling. This produced a delayed time-to-peak of the coupled
membrane potential for synaptic inputs locally activated near the soma of the pre-
coupled neuron. Although a delay is expected for distally located inputs, such a
delay is not physiological for inputs located proximal to the soma in the absence of
any shunts or regenerative membrane currents, since gap-junctions are associated
with purely resistive electrotonic coupling.
The purpose here will be to undertake a more quantitative analysis of electro-
tonic coupling than previously discussed by Poznanski et al. [30]. This will be accom-
plished by deriving more realistic values of the somatic coupling strength parameter
(R), defined as the ratio between the resistances of the somatic membrane (Rs ) to
the somatic gap-junction (Rj ). In general the value of the parameter R used by
Poznanski et al. [30] was too small to reveal the importance of the model in predict-
ing subthreshold activity observed in hippocampal pyramidal neurons. Estimates of
R were selected in the range 0.01 < R < 2, and this has led to a delayed time-to-
peak of the electrotonic potentials for locally activated synaptic input. Earlier work
had shown that coupling ratios between CA3 pyramidal cells to be 0.05 to 0.15,
signifying weak coupling [41], whilst coupling ratios of 0.7 and 0.8 were found based
on published values of somatic coupling conductance and photographs of freeze-
fracture gap-junctions [9]. It is expected that new estimates of the somatic coupling
resistance will be found based on coupling ratio analysis for both weak and strong
coupling strength from electrophysiological data and verified using morphological
approach based on freeze-fracture replicas of gap-junctional connexins.
April 16, 2010 16:28 WSPC/S0219-6352 179-JIN 00236

Estimating Gap-Junctional Conductance Between CA3 Hippocampal Pyramidal Neurons 51

The more detailed compartmental models which presumably take into account
the detailed anatomy-physiological complexities of CA3 cells [25] are not required
for analysis of events that occur at the soma [24]. Therefore the tapered cable model
of CA3 pyramidal neurons used in this paper is sufficient for the purpose of measur-
ing the somatic gap-junctional conductance. Evans [10–12] extended the model by
obtaining a full analytical solution based on soma-somatic coupling [10] and dendro-
dendritic coupling [11] between cables. However, in neither of those papers were the
estimates of the somatic gap-junction resistance parameter (Rj ) determined from
either electrophysiological data or photographs of freeze-fracture gap-junctions [12].
Furthermore, due to the normalization procedure used by Evans, it is not clear how
an estimate of the somatic coupling conductance could be obtained.

2. Estimates of the Somatic Gap-Junctional Conductance

Based on Coupling Ratios
One way of estimating the somatic gap-junctional conductance (1/Rj ) is to compare
with the use of a model, the experimental data based on the dual intracellular
recording technique involving the injection of current through a microelectrode and
measuring the voltage response in the coupled neuron. By adjusting Rj parameter
in the model in way to match the experimentally derived data, a chosen value of
Rj can be found. The dual intracellular recording technique uses the method of
coupling ratios with sharp electrodes penetrating the somata of coupled neurons
(see e.g., [22]). That produces false estimates of the strength of the coupling as well
as unexpected longer delays in the time-to-peak of the coupling potentials. Likewise,
significant change to the time course of both the synaptic and coupling potentials
due to the electrode leak can occur, but the cable analysis takes into consideration
effects of somatic-shunt (see [30]).
A quantitative evaluation of electrotonic coupling requires a measure of the
strength of coupling between pairs of neurons. The most common measure has been
the coupling ratio or the coupling coefficient [36]. It depends on the membrane
potential of the activated (pre-coupled) neuron and the resistance to current flow
through the gap-junction and across the membrane of the electrotonically coupled
neuron. It is simply defined as the ratio between the somatic peak amplitude of the
voltage change in the coupled neuron to that in the activated neuron:
CR = Us/Vs (1)
where Us and Vs correspond to the peak amplitudes of membrane potentials in
the soma of the coupled neuron U and the input neuron V, respectively. A way of
estimating the coupling ratio (CR) is to choose a selection of the somatic coupling
strength (R = Rs /Rj ) values and passive parameters for individual CA3 neurons
obtained from intracellular current clamp recordings with sharp electrodes using
the dual intracellular recording technique [5, 45] and substituted into the model
equations reproduced in the Appendix. The equations were numerically solved and
April 16, 2010 16:28 WSPC/S0219-6352 179-JIN 00236

52 Poznanski et al.

Table 1. Estimates of the coupling ratios (CRs) for several values of R

where K is the rate of tapering of the dendritic cable model (see Appendix
for equations and definition of terms).

K\R 0.5 1.0 2.0 5.0 10.0

−4.75 0.085 0.155 0.234 0.37 0.51

−1.80 0.1 0.14 0.24 0.47 0.63

the results presented in Fig. 2 and summarized in Table 1 for a range of R values not
considered by Poznanski et al. [30]. The general trend is that coupling ratios increase
for large somatic coupling strength values. The results indicate if R increases then
CR also increases [36].
In Fig. 2 it can be observed that membrane potential in the coupled neuron soma
is delayed for small R values while larger R values produce a significantly smaller

Fig. 2. Time course of the normalized voltage (synaptic potential) at the soma of CA3 pyramidal
neuron 1 (Vs), and the normalized voltage (coupling potential) at the soma of CA3 neuron 2 (Us)
as a result of synaptic input (α-function with α = 50, [17]) activated at a proximal distance from
the soma of CA3 pyramidal neuron 1, along a sealed-end finite dendritic cable of electrotonic length
L1 = L2 = 0.9, whose diameter decreases exponentially at a rate determined by the constant [27]:
(a) K1 = K2 = −4.75 and (b) K1 = K2 = −1.8. Results are given for R = 0, 0.5, 1, 2, 5, and 10 with
the somatic-to-dendritic conductance ratio ρ = 3 [5]. Time is normalized in terms of the membrane
time-constant (τm ).
April 16, 2010 16:28 WSPC/S0219-6352 179-JIN 00236

Estimating Gap-Junctional Conductance Between CA3 Hippocampal Pyramidal Neurons 53

delay between the synaptic and coupling potential. For example, if R = 0.5 then a
delay of about 0.5τm occurs, while if R = 10 then a delay of only about 0.1τm exists.
Experimental studies have shown estimates of the coupling ratio (in the steady
state) for CA3 hippocampal pyramidal neurons to range between 0.05 and 0.15 [41]
and generally under 0.3 [22], suggesting the presence of weak electrotonic coupling.
Based on these experimentally derived estimates of the coupling ratio, it can be
seen from the Table 1 that the somatic coupling strength parameter should be less
than 5 as was previously shown by Poznanski et al. [30]. However, since MacVicar
and Dudek [22] had used sharp electrodes, current leakage around the electrode
is expected to reduce the coupling ratio estimates. The electrode “leak” due to
the sharp electrodes, may actually be larger than the coupling. These theoretical
studies suggest that coupling is stronger than before, but to have much validity
new values with tighter seal patch electrodes should be reviewed. The theoretical
modeling presented here can be further modified to include an access resistance (see
[28, 32]) allowing for experimental verification using the dual whole-cell patch-clamp
technique between cultured hippocampal neurons [26] considering large pipettes
with tighter seal are not expected to damage the somatic membrane, thus a reduced
time-to-peak is expected to be evident in the membrane potential of the coupled
neuron soma.
Evans [12] has obtained theoretical estimates of CR through the use of analytical
solutions to the equations for a model of a pair of CA3 hippocampal pyramidal
neurons electrotonically coupled between their somata. Estimates found were CR =
1, 0.8, 0.6 and 0.25 for µ(= RD∞ /Rj ) = 50, 10, 5 and 1, respectively, indicating
a higher coupling strength than previously considered both theoretically [30] and
experimentally [22]. However, Rj estimates cannot be explicitly determined from
CRs due to the normalization of the parameters. Therefore other methods are used
to determine an appropriate somatic gap-junctional coupling conductance.
Poznanski et al. [30] derived the following relation for the somatic coupling
conductance:
Rj = RN {(1 − CR)/CR} (2)
where RN is the input resistance of the coupled neuron (U) in the pair. Note that
CR → 1 when Rj → 0 denotes strong coupling, and CR → 0 as Rj → ∞ denotes
weak coupling. However as mentioned, the junctional resistance based on coupling
ratio analysis is only suitable for weak coupling because it would be difficult to
distinguish the peak somatic potential in the coupled neuron if the coupling was
strong. Further, estimates of the parameter need to be examined with reference to
both strong and weak coupling.

3. Cable Parameter Estimates of the Somatic

Gap-Junctional Conductance
MacVicar and Dudek [22] observed that coupling ratio values are greater than
0.3, but were ignored as possible responses from the same neuron using the dual
April 16, 2010 16:28 WSPC/S0219-6352 179-JIN 00236

54 Poznanski et al.

intracellular recording technique. Given the possibility of strong coupling among

CA3 hippocampal pyramidal neurons exists, the coupling ratio as a quantitative
measure of coupling strength is no longer useful and cable estimates of the somatic
coupling junctional conductance are necessary based on known electrophysiologi-
cal data.
The somatic coupling resistance (Rj ) in MΩ can be given by the following
expression:
Rj = RN [1 + (1 + ρ)/R] (3)
where RN is the input resistance of the coupled neuron (U) in the pair in MΩ, ρ =
Rs /RD is the somatic-to-dendritic conductance ratio (dimensionless), and R = Rs /Rj
is the somatic strength parameter (dimensionless). Under current-clamp, the input
resistance (RN ) of the coupled cell is measured from the slope of the steady-state
voltage–current (V–I) relationship (curves with anomalous rectification should be
excluded from the analysis). This is achieved by injecting current steps intracellu-
larly and measuring the resultant voltage transients (assuming no gated currents are
activated). The only estimate of RN in the range from 2 to 33 MΩ are for electro-
tonically coupled aged CA3 hippocampal pyramidal neurons [1]. These values are
lower than those found by Brown et al. [5] for uncoupled CA3 pyramidal neurons,
suggesting that electrotonic coupling lowers the input resistance of neuron or that
aging alters the cable properties of neurons. The somatic-to-dendritic conductance
ratio (ρ), unlike the input resistance of a neuron, is not changed by the presence of
electrotonic coupling and values obtained from intracellular current clamp record-
ings with sharp electrodes from single CA3 pyramidal neurons (in the absence of
electrotonic coupling) range from 2.4 to 3 [5, 18, 44, 45].
A clearly defined range of Rj values is evident as shown in Fig. 3 using previous R
values (which encompass both strong and weak electrotonic coupling between CA3
pyramidal cells), together with the above values of ρ and RN substituted into Eq. (3).
The choice of parameter values has been given a systematic account of the possible
ranges of ρ and RN for CA3 hippocampal pyramidal neurons [5, 18, 44, and 45], for
example, based on sharp electrodes measurement of CA3 neurons RN = 36–42 MΩ
[5] and ρ = 2.7 [18]. Weak electrotonic coupling expressed by R = 0.5, 1 and 2 shows
large values of the somatic coupling resistance, even possibly approaching the upper
limit for undetectable dye-coupling of 500 MΩ [8]. Strong coupling indicated by R>5
or coupling ratios above 0.3 (see Table 1) show that the somatic coupling resistance
is contained below 100 MΩ, and also there is a small variation in Rj values for the
chosen range of ρ values.
Foregoing experimental work of MacVicar and Dudek [22] appear to show low
coupling coefficients (CRs) in the range of 0.05 to 0.15 for CA3 hippocampal pyra-
midal neurons. From our results this would indicate that the coupling strength
parameter (R) is less than unity, and therefore the coupling coefficient and somatic
coupling resistance can reach high levels depending on the measured input resistance
(RN ), and to a lesser extent, the ρ values.
April 16, 2010 16:28 WSPC/S0219-6352 179-JIN 00236

Estimating Gap-Junctional Conductance Between CA3 Hippocampal Pyramidal Neurons 55

Fig. 3. The somatic coupling resistance (Rj ) plotted against input resistance (RN ) for specifically
selected somatic coupling strength parameter values (R) reflecting both weak and strong electro-
tonic coupling, as well as three ρ values selected from estimates made on single CA3 hippocampal
pyramidal neurons.

These results will clearly need additional verification of the coupling conductance
parameter. The use of a morphological approach based on freeze-fracture replicas of
gap-junctional connexins will be needed to conclusively estimate the somatic gap-
junctional conductance between CA3 hippocampal pyramidal somatas that serves
as a parameter of an ensemble formed by many individual gap-junctions.

4. Morphological Estimates of the Somatic

Gap-Junctional Conductance
A morphological approach can be used to constrain the estimation of the coupling
resistance which we will denote as (Rc ) to distinguish from the physiologically esti-
mated somatic coupling resistance (Rj ). Nevertheless, in principle they both repre-
sent the same parameter. The coupling resistance (Rc ) can be expressed by
Rc = Rchannel /N (4)
where N is the number of open channels in the gap-junction and Rchannel is the
resistance of a single junctional channel. From this relation it can be seen that
the number of particles in a gap-junction determines its strength (i.e., the greater
the number of particles the stronger the coupling via a smaller somatic coupling
resistance). This relationship ignores the modulation of gap-junction coupling by
second-messengers reported between CA3 pyramidal axons [13].
April 16, 2010 16:28 WSPC/S0219-6352 179-JIN 00236

56 Poznanski et al.

Gap junctions are aggregated channels with the number of channels (N) esti-
mated by freeze-fracture electron microscopy, on the assumption that all particles
in plaques, counted from freeze-fracture replicas, are functional gap-junction chan-
nels and are in the open state for most of the time. If not all pores are open in plaques
counted from freeze-fracture then from Eq. (4) smaller N values could reflect greater
Rc values, but extremely high values of Rc would imply undetectable dye coupling.
This is because some dyes (e.g., Lucifer yellow) cannot flow into the coupled cell for
a Rc value greater than 500 MΩ [8], implying that the true extent of coupling could
be greater than those predicted from Lucifer yellow dye-coupling.
Each channel consists of two connexons located in the membranes of adjacent
CA3 pyramidal cells and separated by a small “gap” (see Fig. 4). Each connexon
accommodates six proteins called connexins, thus forming an aqueous pore [2].
Electrical synapses via gap-junctions between hippocampal CA3 pyramidal neurons

Fig. 4. The electrical synapse is a collection of connexin channels that would be seen ultra-
structurally in freeze-fracture replicas as particles in plaques corresponding to gap-junctional
channels. The somatic coupling resistance (Rj ) gives an estimate of the gap-junctional resistance
influenced by an ensemble of unitary resistances of gap-junctional channels Rchannel.
April 16, 2010 16:28 WSPC/S0219-6352 179-JIN 00236

Estimating Gap-Junctional Conductance Between CA3 Hippocampal Pyramidal Neurons 57

might be composed of connexin Cx36 [7, 40]. These studies showed Cx36 mRNA
was particularly prominent in the CA3 region and immune-identification of Cx36
using antibodies was significant between hippocampal pyramidal cells. To estimate
the resistance of a single junctional channel, it is assumed that the channel is cylin-
drical, and consists of the sum of its pore resistance and access resistances of its
mouths [16]:
Rchannel = ri Lchannel /(πa2channel ) + 0.5 ri /achannel (5)
where ri is the resistivity of the intracellular medium (Ωcm), Lchannel is the length
of the pore (nm), and achannel is the channel (pore) radius (nm). For example, if we
assume ri = 200 Ωcm derived from patch-clamp studies of CA3 neurons [38], and
using liver cell gap-junction estimates of achannel = 1 nm (as the average limiting
diameter of the largest dye molecule that can pass through the junctional channel),
together with Lchannel = 15 nm (from electron microscopy) [6], then this gives a
conductance of 1/Rchannel = 95 pS. Equation (5) can also be adapted to deal with
the case of a channel with variable radius [39]. If neuronal gap-junctions are smaller
with achannel = 0.6 nm and Lchannel = 10 nm based on data from the giant axon
of Lumbricus terrestris [4] then this gives a conductance of 1/Rchannel = 50 pS.
Although there is no a priori reason that Rchannel will not vary between different
neurons, the available evidence suggests a range of unitary conductances of gap-
junctional channels between 50 to 150 pS for various connexins [8]. In fact, more
recent data suggest that connexin Cx36 carries a very low unitary conductance
of 10–15 pS [40]. Another assumption is that the unitary resistance of all connxin
channels associated with a grouping of gap-junctions from freeze-fracture replicas
(plaques) is of the same value.
The somatic coupling strength (R) follows from its definition Rs /Rj to equal
8.2 if (i) the somatic membrane resistance (Rs ) is found from membrane resistivity
estimates using the relation: Rs = Rm /surface area of soma where some ad hoc
estimates used by Traub and colleagues [41, 43]: Rm = 10,000 Ωcm2 and soma surface
area 3,320 µm2 [43] give Rs = 301 MΩ, and (ii) the junctional resistance is chosen
as Rc = 34 MΩ which was used by Traub and Wong [43] in their network modeling
based on a unitary conductance of a gap-junction channel of 50 pS, which is relatively
low and might suggest that the cell-to-cell communication is strong. The low value
of the gap junctional resistance (RC ) is based on N value taken from the somata of
a CA3 pyramidal cell.
Figure 5 illustrates the relation as expressed by Eq. (4) for three choices of
1/Rchannel . It is apparent that for small N values (under 50) high Rc values are
expected, which would occur when N < 100 for dendritic gap junctions with a more
realistic estimate to be N = 30 to 50 (note: p = 10−12 ). Furthermore, Dudek et al.
[9] estimated the somatic coupling resistance to be 17 MΩ based on estimates of
a unitary conductance for single gap-junctional channel at 100 pS from embryonic
cells of teleosts [2] and photographs of freeze-fracture gap-junctions on somata of
CA3 pyramidal neurons of approximately N = 590 particles in a single plaque [35].
April 16, 2010 16:28 WSPC/S0219-6352 179-JIN 00236

58 Poznanski et al.

Fig. 5. The somatic coupling resistance (Rc ) plotted against the number of open channels in
gap-junctions (N) was found on the somata of CA3 pyramidal neurons for three values of the
conductance of a single junctional channel 1/Rchannel. The estimate range of N is based on the
assumption that only a small percentage of all pores are in the open state for most of the time.

It may be also possible that not all pores are open in plaques counted from freeze-
fracture during activation of gap-junction or that the unitary conductance of a gap-
junction channel for CA3 hippocampal pyramidal neurons is below 100 pS. Hence
from Eq. (4), smaller N or larger Rchannel values could reflect greater Rj values and
also smaller R values. If we assume N = 590 particles in a single plaque on somata
of CA3 hippocampal pyramidal neurons [9] are open then the somatic membrane
gap-junctional resistance would be under 100 MΩ and the electrophysiological data
indicate strong coupling. However, if we assume that only 1% of particles in plaques
are open gap-junction channels then the somatic membrane gap-junctional resistance
becomes greater than 5000 MΩ, implying weak coupling.
A simple algorithm on how to estimate the somatic coupling junctional resistance
is shown below:

Step 1: Obtain estimates of the CR’s as given by Eq. (1).

Step 2: If CR < 0.5 then go to step 3 else go to step 4.
Step 3: Weak somatic coupling: Rj is given by Eq. (2); go to step 5.
Step 4: Strong somatic coupling Rj is given by Eq. (3).
Step 5: Compare with the morphological estimate Rc given by Eq. (4).
Step 6: If Rj ≈ Rc then end; else go to step 1.

Few comments are needed with regard to the choice of the parameters. Firstly,
estimates of the somatic gap-junctional conductance using sharp microelectrodes will
April 16, 2010 16:28 WSPC/S0219-6352 179-JIN 00236

Estimating Gap-Junctional Conductance Between CA3 Hippocampal Pyramidal Neurons 59

give different estimates to those measuring the transjunctional conductance through

gap-junctions using dual whole-cell patch-clamp. The latter techniques of recording
from coupled neurons limit the leakage caused by a sharp microelectrode penetrating
the somatic membrane. Therefore, there is no need for a somatic-shunt parameter
to be introduced in the model. Secondly, patch-clamp measurements of the passive
properties of CA3 pyramidal neurons show increased values of RN in comparison
with sharp electrode measurements [19]. This would also imply greater somatic
coupling resistance values (see Fig. 3). Indeed if cells with larger RN values also have
a smaller number of gap-junction channels (N) then from Eq. (4) this would imply
greater Rc values, as shown by Bigiani and Roper [3] using whole-cell patch-clamp
recordings from receptor cells in the Necturus taste buds. Thirdly, estimates of CR
values obtained for CA3 hippocampal pyramidal neurons were estimated with sharp
electrodes using a dual intracellular recording technique [22], and therefore a direct
comparison between estimates of CR values obtained for CA3 pyramidal neurons
using sharp electrodes and patch-pipettes cannot be made. The leakage caused by the
sharp electrode is included in the model, however, from the analysis presented here it
is clear that estimates of the somatic coupling strength (R) show that the CR of CA3
pyramidal neurons is greater than previously presented using sharp electrodes [22],
implying a more stronger coupling exists between CA3 pyramidal neurons. Fourthly,
mechanisms of electrical synaptic plasticity were not considered in the model. For
simplicity we assumed an ohmic gap-junctional conductance without reference to
voltage dependence of coupling or to its modulation by various neurotransmitters
and second messengers.

5. Discussion
Coupling ratio analysis involving sharp electrodes may also include components of
regenerative voltage responses generated in coupled systems. Since our model only
included passive membrane the simulation will give misleading results for estimating
coupling properties of electrical synapses even in the sub-threshold range of voltage.
There are two ways to extend the model: firstly, use of models with active membrane
can be utilized based on concept of ionic cable theory (see [12]); secondly, since the
dual whole-cell patch-clamp measures only the transjunctional conductance Rj , it
would avoid the nonlinear effects and therefore is a better technique. Our model can
be adapted to the whole-cell patch clamp with the introduction of the patch-access
resistance [20, 32].
The evidence of somatic coupling between mature CA3 pyramidal neurons comes
from MacVicar and Dudek [21] and Dudek et al. [9] using older recording tech-
niques. There is ultrastructural evidence of somatic electrotonic coupling in guinea
pig hippocampus [35]. It was shown that CA3 pyramidal neurons express soma-
somatic gap junctions, although electrotonic synapses were not proven, and there-
fore gap-junctions on the somata of CA3 pyramidal neurons from that study could
April 16, 2010 16:28 WSPC/S0219-6352 179-JIN 00236

60 Poznanski et al.

function for metabolic communications or cooperation independently of electrical

interactions.
The physiological role of somatic electrotonic synapses were hypothesized to
synchronize electrotonic excitatory postsynaptic potentials by shunting current from
one CA3 pyramidal cell to another in the presence of chemical synaptic activity
[29] in the same way as “ultrafast” communications between CA1 (n = 82) and
to some extent CA3 (n = 7) pyramidal neurons [33]. Further work is required
to determine the functional role of axo-axonal coupling in a cluster or network of
neurons interconnected by axo-axonal gap-junctions [42], as ultrastructural evidence
for axo-axonic gap-junctions has only so far been obtained in dentate granule cell
mossy fibers which are synaptically connected to the CA3 pyramidal neurons [15].

6. Conclusion
We have presented a model that describes electrophysiological properties of signal
transmission between pairs of CA3 hippocampal pyramidal neurons based on elec-
trophysiological experiments for the membrane potential responses. We proposed
a method to estimate the gap-junctional conductance between pairs of somatically
coupled CA3 pyramidal cells, based on simulated membrane potential responses,
which derived from a cable model taking the gap-junction as a linear conductance.
The effectiveness of our proposed method has been confirmed ultrastructurally to
suggest strong somatic coupling between pairs of CA3 pyramidal cells.

Acknowledgement
We thank Professor S. Hidaka for fruitful discussions and comments on this paper.

Appendix
The classical model of a dendritic tree is referred to by a tapered equivalent core-
conductor [14, 27]. A dendritic tree can be represented in terms of a continuously
tapering cable if the following key assumption is satisfied: the dendritic branches are
cylindrical and their diameters at equal electrotonic distance each raised to the 3/2
power approximate an exponential taper with electrotonic distance — the combined
dendritic trunk parameter (Dtaper ) defined by the so-called branching condition:
 


ni 
no 
Dtaper = dj(x)3/2 dj(x)3/2 ≈ exp(2/3Kz)
 
j=1 j=1

where dj represents the diameter of the j-th branch at a given electrotonic distance
from the soma, no is the number of primary trunks emanating from the soma, ni
is the number of branches at any particular distance from the soma. It should be
mentioned that in a tapered modification of the equivalent cylinder, all dendritic
April 16, 2010 16:28 WSPC/S0219-6352 179-JIN 00236

Estimating Gap-Junctional Conductance Between CA3 Hippocampal Pyramidal Neurons 61

branches are not required to terminate at the same electrotonic distance from the
soma. In view of this, the electrotonic length of a tapered equivalent cable (L) should
be interpreted to correspond to the maximum electrotonic length of a particular
branch of the dendritic tree [31].
The basic equations of the problem is a system of convection-diffusion equations
coupled at the boundary (Z = 0). The membrane potential of the pre-coupled neuron
V (in mV) activated by a synaptic current injection at z = −ξ satisfies the following
boundary-value problem:
VT = Vzz − K1 Vz − V + ri,taper λtaper Isyn (T)δ(z + ξ)
(1 + Rs /Rj )V(0, T) + γVz (0, T) + σVT (0, T) = Rs /Rj U(0, T)
Vz (z = −L1 , T) = 0
V(z, 0) = 0.
The membrane potential of the coupled neuron U (in mV) satisfies the following
boundary-value problem:
UT = Uzz + K2 Uz − U
(1 + Rs /Rj )U(0, T) − γUz (0, T) + σUT (0, T) = Rs /Rj V(0, T)
Uz (z = L2 , T) = 0
U(z, 0) = 0
x
where z = dξλtaper is the electrotonic distance (dimensionless); x is the physical
distance from the soma (x = 0) in cm, λtaper = λ exp(Kz/3) is the space-constant
of an exponentially tapering cable (cm); λ = [(Rm/4Ri)D]1/2 is the characteristic
length parameter of a uniform equivalent cable (cm); D being the diameter of a
uniform equivalent cable structure (cm), and the actual tapering of the equivalent
cable structure is given by Dtaper = Dexp(2Kz/3) where K < 0 is a rate constant
which determines the amount of taper in the equivalent cable; Ri is the resistivity of
the intracellular medium (Ω cm); T is the dimensionless time (t/τm ); τm = Rm Cm
is the membrane time-constant (msec); Cm is the membrane capacitance (µF/cm);
Rm is the resistance across a unit are of membrane (Ωcm2 ); Rs = Rm /πd2soma is the
somatic input resistance (MΩ); dsoma is the diameter of the soma(cm); ri,taper is the
intracellular resistivity (Ω/cm); L1 and L2 are the electrotonic lengths of cable in
neuron 1 and 2, respectively; K1 < 0 and K2 < 0 are the tapering rate constant
of cable in neuron 1 and 2, respectively; γ = Rs/ri,taper , λtaper is the somatic-to-
dendritic conductance ratio for a semi-infinite cable (dimensionless); ρ = γ tanh(L)
is the somatic-to-dendritic conductance ratio of a cable with electrotonic length L
(dimensionless); σ = τs /τm is the somatic-shunt parameter (dimensionless); τs =
Rs Cs is the somatic membrane time-constant (msec); Cs is the capacitance of the
somatic membrane (µF/cm); RN = Rs RD /(Rs + RD ) is the input resistance of the
neuron (MΩ); RD = Rs /ρ is the input resistance of the dendritic cable (MΩ); δ is the
Dirac-delta function and subscripts z and T denote partial derivates with respect to
these variables.
April 16, 2010 16:28 WSPC/S0219-6352 179-JIN 00236

62 Poznanski et al.

References
[1] Barnes CA, Rao G, McNaughton BL, Increased electrotonic coupling in aged rat hip-
pocampus: A possible mechanism for cellular excitability changes, J Comp Neurol
259:549–558, 1987.
[2] Bennett MVL, Spira ME, Spray DC, Permeability of gap-junctions between embryonic
cells of fundulus: A re-evaluation, Dev Biol 65:114–125, 1978.
[3] Bigiani A, Roper SD, Estimates of the junctional resistance between electrically coupled
receptor cells in Necturus taste buds, J Gen Physiol 106:705–725, 1995.
[4] Brink PR, Dewey MM, Evidence for fixed charge in the nexus, Nature 285:101–102,
1980.
[5] Brown TH, Frike RA, Perkel DH, Passive electrical constants in three classes of hip-
pocampal neurons, J Neurophysiol 46:812–827, 1981.
[6] Casper DLD, Goodenough DA, Makowski L, Phillips WC, Gap junction structure.
I. Correlated electron microscopy and X-ray diffraction, J Cell Biol 74:605–628, 1977.
[7] Condorelli DF, Parenti R, Spinella F, Trovato-Sanlinaro A, Belluardo N, Cardile V,
Cicirata F, Cloning a new gap junction gene (Cx36) highly expressed in mammalian
brain neurons, Eur J Neurosci 10:1202–1208, 1998.
[8] Dermietzel R, Spray DC, Gap junctions in the brain: Where, what type, how many
and why? Trends Neurosci 16:186–192, 1993.
[9] Dudek FE, Andrew RD, MacVicar BA, Snow RW, Taylor CP, Recent evidence for and
possible significance of gap junctions and electrotonic synapses in the mammalian brain,
in Jasper HH, van Gelder NM (eds.), Basic Mechanism of Neuronal Hyperexcitability
Liss, New York, 1983.
[10] Evans JD, A cable model for coupled neurons with somatic gap junction, Biol Cybern
92:164–176, 2005.
[11] Evans JD, Analytical solution of the cable equation with synaptic reversal potentials
for passive neurons with tip-to-tip dendrodendritic coupling, Math Biosci 196:125–152,
2005.
[12] Evans JD, Multicylinder models for synaptic and gap-junctional integration, in Reeke
GN, Poznanski RR, Lindsay KA, Rosenberg JR, Sporns O (eds.), Modeling in the
Neurosciences: From Biological Systems to Neuromimetic Robotics, 2nd ed., CRC Press,
Boca Raton, FL, 2005.
[13] Gladwell SJ, Jefferys JG, Second messenger modulation of electrotonic coupling
between region CA3 pyramidal cells axons in the rat hippocampus, Neurosci Lett
300:1–4, 2001.
[14] Goldstein SS, Rall W, Changes of action potential shape and velocity for changing core
conductor geometry, Biophys J 14:731–757, 1974.
[15] Hanzei-Sichani F, Kamasawa N, Janssen WGM, Yasumura T, Davidson KGV, Hof
PR, Wearne SL, Stewart MG, Young SR, Whittington MA, Rash JE, Gap junctions
on hippocampal mossy fiber axons demonstrated by thin-section electron microscopy
and freeze-fracture replica immunogold labeling, Proc Natl Acad Sci USA 104:12548–
12553, 2007.
[16] Hille B, Ionic Channels of Excitable Membranes, 2nd ed., Sinauer, Sunderland, 1992.
[17] Jack JJB, Noble D, Tsien RN, Electric Current Flow in Excitable Cells, Clarendon
Press, Oxford, 1975.
April 16, 2010 16:28 WSPC/S0219-6352 179-JIN 00236

Estimating Gap-Junctional Conductance Between CA3 Hippocampal Pyramidal Neurons 63

[18] Johnston D, Passive cable properties of hippocampal CA3 pyramidal neurons, Cell Mol
Neurobiol 1:41–55, 1981.
[19] Jonas P, Major G, Sakmann B, Quantal components of unitary EPSCs at mossy fibre
synapse on CA3 pyramidal cells of rat hippocampus, J Physiol (Lond) 472:615–663,
1993.
[20] Krzyzanski W, Bell J, Poznanski RR, Neuronal integrative analysis of the ‘dumbbell’
model for passive neurons, J Integr Neurosci 1:217–239, 2002.
[21] MacVicar BA, Dudek FE, Dye-coupling between CA3 pyramidal cells in slices of rat
hippocampus, Brain Res 196:494–497, 1980.
[22] MacVicar BA, Dudek FE, Electrotonic coupling between pyramidal cells: A direct
demonstration in rat hippocampal slices, Science 213:782–785, 1981.
[23] MacVicar BA, Dudek FE, Electrotonic coupling between granule cells of rat dentate
gyrus: Physiological and anatomical evidence, J Neurophysiol 47:579–592, 1982.
[24] Major G, Passive cable modelling — A practical introduction, in De Schutter (ed.),
Computational Neuroscience: Realistic Modelling for Experimentalists, CRC Press,
Boca Raton, 2000.
[25] Major G, Larkman AV, Jonas P, Sakmann B, Jack JJB, Detailed passive cable models
of whole-cell recorded CA3 pyramidal neurons in rat hippocampal slices, J Neurosci
14:4613–4638, 1994.
[26] O’Beirne MA, Bulloch GM, MacVicar BA, Dye and electrotonic coupling between
cultured hippocampal neurons, Neurosci Lett 78:265–270, 1987.
[27] Poznanski RR, Membrane voltage changes in passive dendritic trees: A tapering equiv-
alent cylinder model, IMA J Math Appl Med Biol 5:113–115, 1988.
[28] Poznanski RR, Electrophysiology of a leaky cable model for coupled neurons, J Austral
Math Soc B 40:59–71, 1998.
[29] Poznanski RR, Strong coupling can reduce the time-to-peak of EEPSPs: A substrate
for neural synchronization, ICONIP 3:1243–1247, 1998.
[30] Poznanski RR, Gibson WG, Bennett MR, Electrotonic coupling between two CA3
hippocampal pyramidal neurons: A distributed cable model with somatic gap-junction,
Bull Math Biol 57:865–882, 1995.
[31] Poznanski RR, Glenn LL, Estimating the effective electrotonic length of dendritic neu-
rons with reduced equivalent cable models, Neurosci Res Comm 15:69–76, 1994.
[32] Poznanski RR, Umino O, Determination of cable parameters for neurons with gap
junctions, in Poznanski RR (ed.), Modeling in the Neurosciences: From Ionic Channels
to Neural Networks, Harwood Academic Publishers, Amsterdam, 1999.
[33] Schmitz D, Schuchmann S, Fisahn A, Draguhn A, Buhl EH, Petrasch-Parwez E, Der-
mietzel R, Heinemann U, Traub RD, Axo-axonal coupling: A novel mechanism for
ultrafast neuronal communication, Neuron 31:831–840, 2001.
[34] Schuster TH, Gap junctions and electrotonic coupling between hippocampal neurons:
Recent evidence and possible significance–a review, Concepts Neurosci 3:135–155, 1992.
[35] Schmalbruch H, Jahnsen H, Gap-junctions on CA3 pyramidal cells of guinea pig hip-
pocampus shown by freeze-fracture, Brain Res 217:175–178, 1981.
[36] Socolar SJ, The coupling coefficient as an index of junctional conductance, J Membrane
Biol 34:29–37, 1977.
April 16, 2010 16:28 WSPC/S0219-6352 179-JIN 00236

64 Poznanski et al.

[37] Sohl G, Maxeiner S, Klaus W, Expression and functions of neuronal gap junctions, Nat
Revs Neurosci 6:191–200, 2005.
[38] Spruston N, Johnston D, Perforated patch-clamp analysis of the passive membrane
properties of three classes of hippocampal neurons, J Neurophysiol 67:508–529, 1992.
[39] Smart OS, Breed J, Smith GR, Sanson MSP, A novel method for structure-based
prediction of ion channel conductance properties, Biophys J 72:1109–1126, 1997.
[40] Srinivas M, Rozental R, Kojima T, Dermietzel R, Mehler M, Condorelli DF, Kessler
JA, Spray DC, Functional properties of channels formed by the neuronal gap-junction
protein connexin36, J Neurosci 19:9848–9855, 1999.
[41] Traub RD, Miles R, Neuronal Networks of the Hippocampus, Cambridge University
Press, New York, 1991.
[42] Traub RD, Schmitz D, Jefferys JGR, Draguhn A, High-frequency population oscilla-
tions are predicted to occur in hippocampal pyramidal neuronal networks intercon-
nected by axo-axonal gap junctions, Neurosci 92:407–426, 1999.
[43] Traub RD, Wong RKS, Synaptic mechanisms underlying interietal spike initiation in
a hippocampal network, Neurol 33:257–266, 1983.
[44] Turner DA, Segmental cable evaluation of somatic transients in hippocampal neurons
(CA1, CA3, and dentate), Biophys J 46:73–84, 1984.
[45] Wong RKS, Prince DA, Afterpotential generation in hippocampal pyramidal cells,
J Neurophysiol 45:86–97, 1981.