JOURNALOFNEUROPHYSIOL~GY

Vol. 60, No. 4, October 1988. Printed

Spread of Synchronous Firing in Longitudinal Slices
From the CA3 Region of the Hippocampus
RICHARD MILES, ROGER D. TRAUB, AND ROBERT K. S. WONG
Department ofNeurology, Columbia University, New York 10032 and IBM T. J. Watson Research
Center, Yorktown Heights, New York, New York 10598
SUMMARY AND CONCLUSIONS
1. Mechanisms underlying the propaga-
tion of synchronous epileptiform activity in
disinhibited hippocampal slices were exam-
ined in experimental and computer simula-
tion studies.
2. Experiments were performed with lon-
gitudinal slices of the CA3 region. Synchro-
nous firing was initiated by stimulating stra-
tum radiatum fibers in the presence of picro-
toxin. It propagated smoothly and without
decrement at velocities close to 0.15 m/s over
distances up to 10 mm.
3. In elevated extracellular calcium, neu-
ronal firing threshold was increased and syn-
chronous burst firing did not spread. Mono-
phasic excitatory postsynaptic potentials
(EPSPs) were recorded in cells at limited dis-
tances from a stimulus in the presence of 10
mM Ca and picrotoxin. Axonal conduction
velocity, estimated from EPSP latencies, was
several times faster than the spread of syn-
chronous firing.
4. EPSPs recorded in 5-7 mM Ca and pic-
rotoxin could consist of two components.
The properties of the first component were
similar to those of synaptic events recorded
in 10 mM Ca. The second component was of
longer latency and unlike the first component
was suppressed in responses to paired stimuli
at interval 50-300 ms. Recordings from cells
at different distances from a stimulus sug-
gested that the second component spread fur-
ther and more slowly than the first compo-
nent.
5. In computer simulations the CA3 re-
gion was represented by a spatially distrib-
uted network of 9,000 excitatory neurons and
900 inhibitory cells. Individual cells and syn-
apses had properties based on experimental
data. The effects of varying synaptic strength
and connectivity on the spread of activity in
the model was examined.
6. When synaptic inhibition was func-
tional in simulations, firing was restricted to
a single action potential in model cells close
to the stimulus, as in experiments. Synchro-
nous burst firing spread throughout the neu-
ronal array when fast synaptic inhibition was
absent. The velocity of propagation was
slower than conduction in simulated axons
when synaptic contacts made by excitatory
cells were spatially limited. Propagation ve-
locity increased with increases in the spatial
extent of excitatory connectivity.
7. Increasing the threshold of neurons in a
region of the model network reduced the
speed at which synchronous firing spread. In
experiments focal application of y-aminobu-
tyric acid (GABA) elevated neuronal firing
threshold and slowed the propagation of syn-
chrony in a local region.
8. As the strength of synaptic inhibition
was gradually reduced, neuronal activity
spread further and faster through the simu-
lated neuronal network. In experiments
when synaptic inhibition was functional, no
response was evoked in cells sufficiently dis-
tant from a stimulus. As inhibition was grad-
ually suppressed long-latency EPSPs were re-
vealed in distant cells, coincident with the on-
set of burst firing in cells closer to the
stimulus. Later, distant cells discharged
bursts at shorter latencies as synchronous
population activity spread throughout the
slice.
9. We suggest that the spread of synchrony
results from a continuous process in which
active neurons synaptically excite distant in-
OO22-3077/88 $1.50 Copyright 0 1988 The American Physiological Society
1481
1482
R. MILES, R. D. TRAUB, AND R. K. S. WONG
active neurons. Both synaptic inhibition and
neuronal firing threshold affect the spread of
firing via excitatory synaptic pathways. In
this way they influence the extent and speed
with which synchronous firing spreads.
INTRODUCTION
When synaptic inhibition is suppressed
CA3 hippocampal pyramidal cells discharge
in stereotyped, highly synchronous, bursts of
action potentials. This activity has been com-
pared to interictal epileptiform events (13-
1529, 5 1,6 1). A number of mechanisms in-
cluding changes of interstitial ions (32, 50,
52), electrotonic coupling (38, 54), and eph-
aptic interactions (22, 28, 54) may enhance
the synchrony of neuronal firing. However in
the CA3 region of the hippocampus synchro-
nous burst firing appears to be initiated by a
depolarization resulting from transmitter re-
lease at chemical synapses (5, 14,23, 29,41).
Recurrent excitatory synapses between CA3
pyramidal cells may be responsible (5, 35,
37). The observation that activity in a single
CA3 neuron can influence synchronous fir-
ing of the CA3 cell population supports this
assertion (39). A reduction of synaptic inhibi-
tion may enhance the spread of activity
through polysynaptic recurrent excitatory
pathways (9,43,44).
If mechanisms for the initiation of interic-
tal activity in a focus are becoming clearer
( 14), questions remain concerning the spread
of seizure activity from a focus. It is known
that focal discharges may induce neuronal
synchronization in distant, synaptically con-
nected regions (12, 48, 55) although the
mechanisms underlying this form of plastic-
ity are unclear. A related problem is to iden-
tify factors that control the spread of activity
in spatially distributed neuronal networks
(20). In this report we examine the spread of
synchrony within a large disinhibited cell
population using experimental and computer
simulation techniques.
Synchronous firing is often initiated in the
CA3 region of transverse hippocampal slices
(23,3 1,4 1,5 1). In these slices CA3 pyramidal
cell bodies are arranged in a curved layer up
to 3 mm long. In the present study slices were
prepared from the CA3 region cut in the lon-
gitudinal septal-temporal plane of the hippo-
campus. Longitudinal slices had several ad-
vantages for our experiments. The CA3 cell
body layer was straight and up to 10 mm
long, thus offering an improved spatial reso-
lution and reducing uncertainties in measur-
ing distances within a curved structure. Most
afferent fiber systems were probably cut in
preparing these slices. We could then assess
the role of recurrent connections between
neurons in a large population of similar cells
in the spread of synchronous firing.
Mechanisms controlling the propagation
of synchronous discharges were also assessed
with a computer model of the CA3 region
(56-59). In this study the number of simu-
lated neurons was increased to compute the
spread of activity in a large spatially distrib-
uted cell population. Active currents and pas-
sive properties of excitatory and inhibitory
cells were based on experimental data from
CA3 hippocampal cells (56). Synaptic prop-
erties were modelled on those of excitatory
and inhibitory connections between CA3
cells (37, 40, 42). Little information is avail-
able on synaptic connectivity-the number
and spatial distribution of synapses made by
single CA3 cells (9, 16,25,26,53). The ability
to assess in simulations the effects of varying
synaptic strength, density, and spatial distri-
bution was especially useful.
We found that when inhibition was sup-
pressed, synchronous firing spread through-
out longitudinal slices. Synchrony spread
more slowly than conduction in individual
axons. Simulations suggested that a spatially
restricted pattern of excitatory synaptic con-
nectivity could account for this observation.
Both synaptic inhibition and neuronal
threshold were found to control the spread of
synchronous firing.
METHODS
Experimental techniques
Guinea pigs weighing 200-300 g were used.
Their forebrain was rapidly removed after cervical
dislocation and one hippocampus was isolated.
The curvature of the hippocampus was reduced
when its medial surface was stuck to a block with
cyanoacrylate glue. The block was placed in a tis-
sue slicer and longitudinal slices were cut in a
plane orthogonal to the CA2-3a hippocampal sur-
face. One or two longitudinal slices, of thickness
400 pm and length 8-10 mm, included the CA3
region. In preliminary experiments longitudinal
slices were stained with methylene blue. Cell bod-
SPREAD OF SYNCHRONY IN HIPPOCAMPUS 1483
ies in the stratum pyramidale were seen to extend
the length of the slice. Next to it the stratum luci-
dum (mossy fiber layer) was observed as a distinct
band confirming that slices were from CA3 and
not from the CA1 region. Both layers were visible
in transilluminated unstained slices from which
electrical recordings were made.
Slices were transferred to a recording chamber
where they were supported on nylon mesh. They
were exposed to a warmed, moistened 5% CO;! in
O2 atmosphere at 37C. Their lower surface was
perfused with a solution containing (in mM)
NaCl, 124; KCl, 4.5; CaCl*, 2- 10; MgC12, 2;
NaHC03, 26; and d-glucose, 10 at a pH of 7.4.
Picrotoxin ( lo- 100 PM) was usually added to sup-
press Cl-dependent synaptic inhibition mediated
by y-aminobutyric acid (GABA). In some experi-
ments GABA (100 PM) dissolved in perfusing so-
lution was focally applied by pressure ejection
from low resistance glass electrodes (Picospritzer,
General Valve).
Field potentials were recorded using electrodes
of resistance 5- 10 MQ which contained 2 M NaCl.
Intracellular potentials were recorded with elec-
trodes filled with 3 M K-acetate, bevelled to a resis-
tance of 50-70 MQ. Recordings were made from
the stratum pyramidale with electrodes controlled
by separate manipulators (Zeiss, Jena). Signals
were amplified by high-input impedance amplifi-
ers (WPI M707). Fibers in the stratum radiatum
were stimulated with bipolar tungsten electrodes
using electrical pulses of duration 50- 100 ps. Sig-
nals were displayed on a digital oscilloscope (Nico-
let 4562) and recorded on FM tape (Vetter). Some
synaptic responses were averaged and electrode
coupling artefacts were digitally suppressed (42).
In some experiments synaptic responses in cells
at varying distances from a fixed stimulating elec-
trode were examined. Distances were measured
with an accuracy of - 10% using small strips of
graph paper placed close to the slice. Stimulus in-
tensity was adjusted to elicit a maximal response
in an extracellular recording made at a distance of
1 mm. The maximal distance from the stimulus at
which a field potential could be detected was deter-
mined. Intracellular recordings were then made
from cells at distances within and just beyond this
range. They were usually made in random order.
Records were obtained from more than one cell
at the same distance from the stimulus in some
experiments to check that synaptic responses were
reproducible.
Computer simulations
Computer programs were written in FOR-
TRAN, run on an IBM 3090 machine with 16
Mbytes of virtual memory utilizing the vector fea-
ture. The simulated neuronal network comprised
9000 pyramidal (E) cells and 900 inhibitory (I)
cells (45). An array of 225 X 20 X 2 pyramidal cells
was constructed. Distances between cells in the ar-
ray were equivalent to 20 pm, so the 225 rows of
the array represented a longitudinal distance of
4500 pm. Each row was 2 cells wide, correspond-
ing to 2 pyramidal cells at each point within the
stratum pyramidale. Rows were 20 pyramidal cells
deep, corresponding to a slice 400 pm thick. Each
row also contained 4 I cells. Neuronal activity was
often summed over blocks of 15 rows, each equiv-
alent to 300 pm in length (Fig. 1).
CELLULAR PROPERTIES. Pyramidal cells were
modelled with 28 compartments representing
soma with apical and basilar dendrites. Active
conductances, representing Na, Ca, delayed recti-
fier and Ca-dependent K currents (56), were re-
stricted to the somatic compartment. Passive
properties- membrane and cytoplasmic resistiv-
ity, specific capacitance and leakage conduc-
tance-were modelled as previously (57, 60). A
simulated depolarizing current injection elicited a
burst of action potentials followed by a hyperpo-
larization.
In an attempt to account for the fast and slow
phases of synaptic inhibition elicited in CA3 cells
by afferent stimulation, two types of inhibitory
cells were simulated (59). It has been shown that
unitary inhibitory postsynaptic potentials (IPSPs)
do not have uniform properties and that more
than one type of inhibitory cell exists. In the CA1
region repetitively firing inhibitory cells (30, 33)
initiate IPSPs which differ from many spontane-
ously occurring events (2, 10). In CA3 two types of
inhibitory cells initiate unitary events of differing
amplitude and time course (40, 59). The reversal
potential and picrotoxin sensitivity of the larger,
fast, unitary IPSPs suggest they are Cl-dependent
(40,43) and no late, slow postsynaptic component
appears when presynaptic cells fire maximally.
However, the correlation of activity in different
classes of inhibitory cells with the different phases
of the afferent IPSP remains to be conclusively
demonstrated.
In simulations half of the inhibitory cells initi-
ated fast IPSPs mediated by Cl and sensitive to
convulsants (43). These cells possessed intrinsic
properties similar to those of pyramidal cells.
Other inhibitory cells elicited convulsant-resistant
IPSPs of slower time course. They did not possess
Ca or Ca-dependent K currents and fired repeti-
tively in response to simulated depolarizations.
SYNAPTIC PROPERTIES. Connections between
model cells represented single functional synapses.
In simulations the properties of four classes of syn-
apses were based as far as possible on experimental
data (8, 10, 30, 33, 37, 40, 42). Pyramidal cells
made excitatory synapses onto other pyramidal
cells and onto inhibitory cells. The two types of
1484 R. MILES, R. D. TRAUB, AND R. K. S. WONG
inhibitory cells made synapses onto pyramidal
cells and onto each other (45, unpublished obser-
vation). Postsynaptic potentials (PSPs) were elic-
ited when presynaptic cells were depolarized >20
mV from rest. Synapses were not activated at inter-
vals shorter than 3 ms to account for axonal refrac-
toriness. Synaptic conductances added linearly
and no attempt was made to model either use or
voltage dependence for synaptic events.
Excitatory synapses onto pyramidal cells gener-
ated a current which reversed at a potential 60 mV
depolarized from rest. The current was modelled
as an alpha function which decayed with time con-
stant 3 ms (8, 59). The peak conductance, Ce, was
15 nS and was distributed to four dendritic sites.
The corresponding somatic conductance was -3
nS (cf. Ref. 42). With this synaptic strength a burst
of action potentials in one cell caused a postsynap-
tic cell to fire when inhibitory inputs were not acti-
vated.
Unitary excitatory synaptic events in repeti-
tively firing inhibitory cells have a faster time
course and larger amplitude than those in pyrami-
dal cells (30,33, unpublished observations). Simu-
lated currents at these synapses reversed at 60 mV
depolarized from rest. Excitatory conductances
followed an alpha function with time constant 1
ms and the peak conductance, Ce, was 12 nS. With
this conductance located at the soma, presynaptic
action potentials elicited postsynaptic spikes with
delay 2-4 ms.
Postsynaptic currents generated by burst-firing
inhibitory cells reversed at 15 mV hyperpolarized
from rest. The peak conductance, Cif, was attained
at 2 ms and it then decayed with time constant 7
ms ( 10). The value of Cif was varied between 0 and
12 nS in different simulations and it was distrib-
uted to 3 postsynaptic sites around and on the
soma.
Repetitively firing inhibitory cells generated
postsynaptic currents which reversed at 25 mV hy-
perpolarized from rest. One presynaptic spike elic-
ited a postsynaptic conductance, Cis, which
reached a peak at 40 ms and declined with time
constant 100 ms. Slow inhibitory currents im-
pinged on the same dendritic compartments as ex-
citatory synaptic currents.
SYNAPTIC CONNECTIVITY. Parameters needed
to define synaptic connectivity for one cell in-
clude: 1) the number of cells with which it forms
synapses, 2) the nature of postsynaptic cells, and
3) the spatial distribution of postsynaptic cells.
Previously (59) values for the first two parameters
were based in part on data from paired recordings
from CA3 cells, separated by ~400 pm in trans-
verse slices. Recordings have been made from 65
cell pairs separated by similar distances in longitu-
dinal slices (44, unpublished observations). Multi-
synaptic inhibitory (n = 17) and excitatory (n = 7)
interactions resembled those observed in trans-
verse slices. We do not conclude from this data
that local synaptic connectivities (within 400 pm)
differ in transverse and longitudinal slices. The
density of synapses made by excitatory and inhibi-
tory cells was assumed to be highest close to the
presynaptic cell and to decay with distance.
The neuronal network was constructed for each
simulation with probabilities that two cells were
connected depending on their type (E- or I-cell)
and spatial location in the array. An average cell
made the following connections. An excitatory
cell made 22 synapses, 20 onto other excitatory
cells and 2 onto inhibitory cells. Each inhibitory
cell made 220 synapses, 200 onto excitatory cells
and 20 onto other inhibitory cells. A typical cell,
excitatory or inhibitory, received 20 excitatory
and 20 inhibitory inputs. These connectivities are
similar to those used in previous simulations.
The probability that two cells were connected
was assumed to decay exponentially with the lon-
gitudinal distance between them. The probability
that an excitatory cell at position, LI, in the array
made a synapse with a cell at position, L2, was
given by P(L,) X [exp - (L, - L2)/XJ. The proba-
bility of connection decayed with space constant
X, and P(L,) was a factor which maintained a con-
stant number of inputs (n = 20) to each cell. With
a space constant of 30 cell diameters for excitatory
connections, one pyramidal cell made 10 of its 20
connections to cells in rows <20 cells, or 400 pm,
away. The number of available cells within this
space was 800, which gave a probability of 1.25%
that 2 cells were directly connected. In simulta-
neous recordings, within this distance in trans-
verse slices, monosynaptic connections have been
detected with a probability of -2%.
The spatial distribution of inhibitory synapses
was varied independently. The probability that an
I-cell at position LI made a synapse with a cell at
position L2 was given by Q(L,) X [exp - (L, - LJ/
Xi]. Xi was the space constant for decay of probabil-
ity of inhibitory connection and Q(L,) was again a
scaling factor. In simulations X, was varied be-
tween 8 and 500 cell diameters ( 160- 10,000
pm) and Xi between 6 and 30 cell diameters (120-
600 pm).
For excitatory synapses a delay dependent on
the longitudinal distance between connected cells
was included. It was scaled to an equivalent con-
duction velocity of 0.5 m/s (6, Fig. 3). No conduc-
tion delay was included for inhibitory connec-
tions.
EXAMINATION OF THE VALIDITY OF THE
MODEL. Many assumptions were made in con-
structing this simulated neuronal network. Their
validity was tested by examining responses of neu-
rons in the array to stimulating cells at one end of
the array. Simulated responses were compared to
SPREAD OF SYNCHRONY IN HIPPOCAMPUS
A
1485
15cells =3OOym 15 blocks l 4.5mm 01 5
Distance mm
A
:;f i =
I 50 nS
I 50 nS
gis -
1100 nS
5omsec
FIG. 1. Comparison of experimental and simulated synaptic responses. A: simulated neuronal array was divided
into 15 blocks of 600 pyramidal cells. Each block corresponded to a longitudinal distance of 300 pm. B: diagram of
experimental arrangement. Synaptic responses were recorded from cells in stratum pyramidale (S.p.) at 3 distances
from a stimulus to stratum radiatum. C: a single action potential was evoked in a cell close to the stimulus site (0
mm). An EPSP was followed by an IPSP with fast and slow phases in a cell at 1 mm and no response was detected in
a cell at 5 mm. D: simulated responses from cells at equivalent positions in the array (blocks 1, 4, and 15). Traces
represent membrane potential, I, , excitatory, g,, fast inhibitory, gif, and slow inhibitory, gi, 7 conductances. A depo-
larizing current stimulus of 3 nA, 2 ms was applied to all cells in block 1 at time t = 0.
synaptic events recorded in cells at different dis-
tances from a stimulus in longitudinal slices (Fig.
1). In this simulation the space constant for con-
nections made by pyramidal cells, X,, was 30 cell
diameters and the space constant for inhibitory
connections, Xi, was 6 cell diameters.
Intracellular responses to stratum radiatum fi-
ber stimulation in longitudinal slices varied with
the distance of the cell from the stimulus. Three
types of response were observed (Fig. 1 C). First, in
cells close to the stimulus a single action potential
was elicited, succeeded by a hyperpolarization. As
shown in Fig. 1D simulated cells at the site of stim-
ulation in the array discharged a single action po-
tential followed by a hyperpolarization dependent
on cellular and synaptic currents. Second, in cells
further from the stimulus (1-4 mm), synaptic re-
sponses consisted of a small excitatory component
followed by fast and slow phases of synaptic inhi-
bition. The simulated synaptic response and un-
derlvina excitatorv and inhibitory synaptic con-
ductances for a cell located at an equivalent posi-
tion in the array are shown in Fig. 1D. Finally, no
response was evoked in cells located ~4-5 mm
from the stimulus site in the slice. In the simula-
tion no response was apparent in cells at the far
end of the array. Thus in a test independent of the
construction of the model, reasonable agreement
was observed between simulated and experimen-
tal synaptic responses.
RESULTS
Spread of synchronized activity in
disinhibited slices
In the presence of picrotoxin ( lo- 100 PM)
synchronous neuronal bursts were initiated
by stimulating fibers in the stratum radiatum
of longitudinal slices. Neighboring neurons
in longitudinal slices fired bursts simulta-
neously with an extracellular field Dotential
1486 R. MILES, R. D. TRAUB, AND R. K. S. WONG
(Fig. 2A). Bursts of action potentials were also
recorded in neurons separated by several mil-
limeters but their latency differed (Fig. 2B).
The latency difference increased with cell sep-
aration. This suggested that synchronous fir-
ing did not occur simultaneously throughout
the slice. Field potential recordings from
multiple locations showed that synchronous
population firing propagated in a wavelike
fashion (Fig. 2C). Propagation velocities (Fig.
20) were quite uniform through each slice.
The mean velocity of field potential propaga-
tion was 0.14 t 0.04 (SD) m/s (n = 17 slices).
There was no systematic decrement in field
potential amplitude as population activity
spread for distances up to 10 mm (Fig. 2C).
This contrasts with the limited region in
which synaptic responses could be detected
when inhibition was functional (Fig. 1 C).
Spatial distribution of monosynaptic
excitatorypostsynapticpotentials (EPSPs)
To assess synaptic contributions to the
spread of synchrony we examined the spatial
distribution of cells in which monosynaptic
EPSPs were evoked by fiber stimulation. In-
hibition was blocked with picrotoxin. Cal-
cium was increased from 2 to 10 mM to re-
duce firing and so suppress polysynaptic in-
teractions (7,27,43). Neuronal threshold was
A
C
6 t
6-h
e L
B
6 1
9 !
mm
2Omsec
elevated by 11 t 4 (SD) mV in 5 cells by this
increase in Ca (17, 24). Synchronous burst
firing did not occur and EPSPs could be ex-
amined in isolation. They did not possess dis-
crete late components and were not de-
pressed in responses to two stimuli at interval
50-300 ms.
Sequential recordings were made from
cells at different distances from a fixed stimu-
lus to examine the dependence of EPSPs on
distance. The spatial properties of these
EPSPs (Fig. 3A) suggested that they could not
underly the propagation of synchrony. First,
they spread more quickly (Fig. 3B). The la-
tencies of EPSPs recorded from cells in nine
slices were used to estimate a conduction ve-
locity for presynaptic axons (Fig. 3B). The
mean conduction velocity was 0.48 t 0.10
m/s. Second, EPSPs were not detected in cells
throughout the slice (Fig. 3A), whereas syn-
chrony invariably spread throughout longitu-
dinal slices. In three slices the peak conduc-
tance of EPSPs in cells at different distances
was estimated from their voltage dependence
and neuronal input resistance (19, 40). In
each case the excitatory conductance de-
clined with increasing distance between cell
and stimulus (Fig. 3C). In no slice was a syn-
aptic response detected in a cell further than
4 mm from the stimulus. However in record-
mm
2GLec
D
007
+
$i
/
+
/
E
/
40-
c
* 7
i+
s 0
4
/
d
/
A
o- J
I 1
1
I
5 10
Distance mm
FIG. 2. Spread of synchronous firing. A: In 50 PM picrotoxin s. radiatum stimulation evoked a synchronous burst
in two cells and an extracellular field potential, e. Simultaneous recordings were made 6 mm from the stimulus and
within 400 pm of each other. B: bursts evoked in simultaneously recorded cells at 6 and 9 mm from the stimulus in
the same slice. C: variation of field potentials with distance from the stimulus in a different slice. D: conduction
velocity for field potential propagation in this slice. One electrode was maintained at 1 mm from the stimulus, and
another electrode recorded potentials sequentially at sites from 1 to 10 mm. The mean and standard deviation (n =
12) of the interval between the onset of field potentials at the two sites are plotted against separation. The conduction
velocity derived by a linear regression fit to these points (broken line) was 0.12 m/s.
SPREAD OF SYNCHRONY IN HIPPOCAMPUS 1487
--II
2.5 -+---------- C
0.5
mm
20 msec
10-I
n=8998753
I
1OmV
2
40mV
I
0
I 1
2 4
Distance mm
0 J -0
I I 1
0.5 2.5 4.5
Distance mm
FIG. 3. Variation of monosynaptic EPSPs with distance. A: EPSP amplitude diminished in cells more distant from
a fixed stimulus. Each trace was averaged from 8 responses recorded in 50 PM picrotoxin and 10 mM Ca. No response
was detected in a cell 4.5 mm from the stimulus. However, moving the stimulus closer to the same cell restored a
response (lowest trace). B: EPSP latency plotted against distance from stimulus to recorded cell. Mean latency and
its standard deviation are shown for n cells from recordings in 9 slices. EPSPs were not detected further than 4 mm
from the stimulus in any slice. The velocity of conduction in presynaptic fibers derived from linear regression (broken
line) was 0.48 + 0.08 m/s. C: variation of peak EPSP conductance with distance in one slice. Conductance was
estimated from the input resistance and the potential dependence of EPSP amplitude measured as shown in the inset.
ings made from distant cells where no synap-
tic response was detected, EPSPs were re-
stored by moving the stimulating electrode
closer to the recorded cell (Fig. 3A). These re-
sults suggest that the extent of axons excited
by stimulation at one site was limited com-
pared to the length of these slices. Note that
this extent was similar to the maximal dis-
tance at which synaptic responses were de-
tected with functional synaptic inhibition
(Fig. 1 C).
Polysynaptic EPSPs spread further than
monosynaptic EPSPs
We next asked whether polysynaptic
EPSPs could mediate the spread of syn-
chrony. If CA3 cells could fire, recurrent syn-
apses between them could generate polysyn-
aptic events. These EPSPs might sustain se-
quential polysynaptic activation of neurons
that were not excited by the initial fiber stim-
ulus.
Inhibition was again suppressed with pic-
rotoxin and calcium increased to levels (5-7
mM) at which hyperpolarization could sup-
press burst generation in recorded cells (n = 5
slices). Single stimuli elicited EPSPs in more
distant cells, up to 7 mm, than in 10 mM Ca.
In an attempt to discriminate between mono-
and polysynaptic EPSPs double stimuli sepa-
rated by 100-300 ms were applied (Fig. 4).
At this interval, burst afterhyperpolarizations
probably render CA3 cells refractory and so
prevent the generation of polysynaptic recur-
rent EPSPs.
Figure 4A shows synaptic responses to two
stimuli at interval 150 ms, from cells re-
corded at different distances. The second
stimulus elicited monophasic EPSPs whose
peak amplitude declined with distance be-
tween stimulus and recorded cell in a similar
fashion to the EPSPs recorded in 10 mM Ca
(Fig. 3A). They were presumed to be medi-
ated monosynaptically. The first stimulus
1488 R. MILES, R. D. TRAUB, AND R. K. S. WONG
5
mm
50 msec
I
20mV
0.5 5
Distance mm
I
20mV
FIG. 4. Spatial distribution of mono- and polysynaptic EPSPs. A: recordings made sequentially from cells at differ-
ent distances from a fixed stimulus (50 PM picrotoxin, 6 mM Ca). Double stimuli at interval 150 ms were applied at
1 Hz and cells were hyperpolarized to prevent firing. The second stimulus evoked synaptic events which declined
with distance and were not detected beyond 3 mm. The first stimulus evoked synaptic responses in cells up to 5 mm
distant. Early and late components of these responses were apparent in cells at distances 2 and 3 mm. B: isolation of
the late component of synaptic events by subtracting the second response from the first response of the cell at 2 mm.
evoked larger EPSPs than the second stimu-
lus in cells at all distances. This suggested that
both mono- and polysynaptic events might
contribute to the first response. A presumed
polysynaptic EPSP was isolated by subtract-
ing the second from the first response. Figure
4B shows the subtraction process for the
EPSPs from the cell at 2 mm from the stimu-
lus where two components were evident in
the first EPSP.
The activation of synaptic or intrinsic K
currents following the first EPSP could have
contributed to the reduction of the EPSP
evoked by the second stimulus. The second
EPSP should then have been most strongly
depressed when the first EPSP was largest.
Since this was not the case, the reduction of
the second EPSP by K currents was probably
small. Instead EPSP depression was most
strongly correlated with distance from the
stimulus to the recording site.
The amplitude and latency of both compo-
nents of synaptic events from this experiment
is plotted against distance from recorded cell
to stimulus site in Fig. 5. The polysynaptic
component was recorded in more distant
cells and spread more slowly than the mono-
synaptic EPSPs. EPSPs were compared with
synchronous burst propagation in the same
slice when Ca was reduced to 3.5 mM. After
1 h records were made from another set of
cells at various distances from the same stim-
ulus site (Fig. 5A). The latencies of synchro-
nous bursts, measured from stimulus to the
first action potential for 16 events in each cell,
are plotted in Fig. 5D. The similarity in con-
duction velocities suggests that the polysyn-
aptic component of synaptic events may me-
diate the spread of synchronous burst dis-
charges.
Simulations of the spread of synchrony
We next examined the spread of neuronal
activity in a simulated neuronal network. As
suggested by experiments, the spatial extent
of excitatory connections was limited com-
pared to the extent of the array (cf. Fig. 3).
Burst firing could spread between connected
cells (43) and fast synaptic inhibition was
suppressed.
In the simulation of Fig. 6 the space con-
stant for the distribution of synapses made by
one excitatory cell, X,, was 30 cells and the
longitudinal extent of the array was 225 cells.
Thus an average cell made 95% of its connec-
tions within 90 cell diameters. On stimulating
cells at one end of the array a nondecrement-
ing wave of activity spread throughout the
SPREAD OF SYNCHRONY IN HIPPOCAMPUS 1489
5mm
t
1
I I I
2 3 4
Distance m m
1
5
4OmV
I I I 1
7 2 3 4 5
Distance m m
FIG. 5. Comparison of synaptic events with propagation of synchrony. Recordings from the same slice as in Fig. 4.
A: synchronous bursts recorded in cells 1, 3, and 5 mm from stimulus (50 PM picrotoxin, 3.5 mM Ca). B: EPSPs
recorded in cells at 1,3, and 5 mm from stimulus (50 PM picrotoxin, 6 mM Ca). These are the same cells as in Fig. 4.
C: amplitude of presumed mono- (0) and polysynaptic (0) components of EPSPs plotted against distance between
stimulus and cell. The components were isolated as in Fig. 4B. D: latency of mono- (0) and polysynaptic (0) compo-
nents plotted for cells at different distances. Filled squares show the latency of the first action potential of the synchro-
nous burst from cells recorded in 3.5 mM Ca (as shown in A). Data points of C and D represent mean and standard
deviation calculated from 16 events.
A
se*
B
600
0
1
:
20 msec
13
I
4
40mV
A poonS
s 2Omsec
C
40
1
E
20-
2
al -
4
O-
Block
FIG. 6. Simulated spread of synchronous firing. A: number of cells firing in blocks 1,4,7, 10, and 13 plotted against
time. At 2 ms a depolarizing current stimulus was applied to all cells in block 1 at time t = 0. B: membrane potential
and excitatory synaptic conductance traces for representative cells from blocks 4 and 13. C: latency of synchronous
activity plotted against block location. Synchronous firing in a block was defined to begin when 50 of its 600 cells
were firing. This level is indicated by the broken line in the lowest trace of A. The equivalent conduction velocity
estimated by linear regression (broken line) was 0.16 m/s.
1490 R. MILES, R. D. TRAUB, AND R. K. S. WONG
network (Fig. 6A). Figure 6B shows traces for
membrane potential and excitatory synaptic
conductance for two cells from different posi-
tions in the array. In this case the velocity
with which population activity spread
through the array was equivalent to 0.16 m/s
(Fig. 6C). The conduction velocity for axons
in the array was 0.5 m/s.
The influence on propagation velocity of
varying the spatial distribution of excitatory
connections was examined (Fig. 7). The
number of synapses made by each cell was
held constant and their spatial distribution
was varied by changing X,. Conduction ve-
locity increased moderately as the spatial ex-
tent of connections was increased. When X,
was 500 cell diameters the probability that a
cell was connected with any other cell in the
array was almost uniform. Then synchro-
nous activity spread at a velocity of 0.48 m/s,
approaching axonal conduction velocity.
Local synaptic integration
synchronized activity
and the spread
of
Both in longitudinal slices and in simu-
lated networks with local connections, popu-
lation activity spread more slowly than axo-
nal conduction velocity. This suggested that
delays in addition to those for conduction
and transmitter release were involved in the
propagation of synchrony. If polysynaptic
pathways formed the substrate for the spread
of synchrony the interval between onset of an
EPSP and the time when the synaptic depo-
larization reached threshold would provide
an additional delay. Increasing neuronal fir-
ing threshold or reducing excitatory synaptic
strength would prolong this synaptic integra-
tion process and so might slow the spread of
synchrony.
In an attempt to elevate neuronal thresh-
old, we applied GABA which in the presence
of picrotoxin should activate postsynaptic
potassium channels on hippocampal neurons
(46). Its effect on the propagation of syn-
chrony was examined (n = 6 slices) using
three extracellular electrodes. They recorded
field potentials close to the stimulus, close to
the site of focal GABA application and at a
distant site. Figure 8A shows field potentials
before and after GABA application. The con-
trol conduction velocity measured between
the most distant recording sites was 0.16 m/s.
When GABA was applied the field potential
recorded close to the application site was
smooth suggesting that cells in this area did
not fire. The conduction velocity between the
most distant sites was reduced to 0.05 m/s.
l
l
l
l
l
l
l
m
0 1
I 4 I
0 50 160 500
Xe
Cells
FIG. 7. Spatial extent of excitatory connections affects the speed at which synchrony spreads. Simulations per-
formed as in Fig. 6 with values of X, between 8 and 500 cell diameters. In the inset the probability that a cell in block
8 was connected to cells throughout the array is plotted for X, values of 15 and 100. The propagation velocity of
synchrony increased with the spatial extent of excitatory connections. When connectivity was essentially uniform,
(X, = 500) the conduction velocity of synchronous firing, 0.48 m/s, approached that of individual connections.
SPREAD OF SYNCHRONY IN HIPPOCAMPUS 1491
Stim
s.p.
&ABA
Control
01 3 --6
Distance mm
3/ -
Block 1
Block5
\
Block 9
n Block 11
\
600
1
Block 15
0
t 20 msec
D
40
E
x20
I
I,/
1 1 I 1
1 5 10 15
Block number
E
11
A
I
-1
20msec
201
! I---
10
ca
0
I
40mV
200nS
I,
r 4 i0 lb
Block number
FIG. 8. Focal GABA application slows the propagation of synchrony. A: field potentials recorded at distances of 1,
3, and 6 mm from the stimulus before and after focal application of 100 PM GABA at 3 mm. This experiment was
simulated by applying a maintained current to prevent firing in blocks 6- 10 of the array. B: number of cells firing in
blocks 1,5,8, 11, and 15 after a stimulus applied to one cell in block 1. C: membrane potential and excitatory synaptic
conductance for simulated cells from blocks 5 and 11. The excitatory input to cell 11 is smaller and rises more slowly
than that for cell 5. D: latency of synchronous firing plotted against block number. The onset of synchrony in a
particular block was defined to be when 10% of its cells were firing (lower broken line in upper trace ofB). E: synchrony
developed most slowly in block 11, distal to the hyperpolarized region. The interval, T,, between firing in 10% and
85% of cells (upper broken line in upper trace of B) is plotted for each block.
The effect of increased neuronal threshold
on the spread of synchrony was examined in
simulations. A conductance increase, equiva-
lent to the opening of channels controlled by
GABA, was applied to prevent firing in cells
of a central portion of the simulated array.
Figure 8B shows the number of cells firing at
several locations in the array including one
where current was applied. Membrane poten-
tial and excitatory conductance traces are
shown for two cells from blocks proximal and
distal to the site of hyperpolarization (Fig.
8C). The excitatory synaptic input to the cell
distal to the site of hyperpolarization in-
creased more slowly than that in the proximal
cell. Population activity also built up more
slowly at sites distal to the hyperpolarized re-
gion (Fig. 8, B and E). Distal cells here re-
ceived fewer synaptic inputs, since some of
their presynaptic cells could not fire. The pro-
longed interval between the onset of synaptic
events and firing in these cells reduced the ve-
locity with which population firing spread
through the arrav (Fin. 80).
Influence of synaptic inhibition on the spread
of synchrony
Finally we examined the influence of syn-
aptic inhibition on the spread of synchrony.
Field potentials recorded during picrotoxin
application suggested that partly synchro-
nous neuronal activity might spread to some
extent before synaptic inhibition was com-
pletely suppressed (Fig. 94. In simulations
the effects of varying the conductance of fast
IPSPs, gif, on the spread of population activ-
ity through the array were examined (Fig. 9,
B and C). With a conductance of 10 nS, stim-
ulation at one end of the array induced firing
that was restricted to cells close to the stimu-
lus site. When the strength of synaptic inhibi-
tion was reduced to 2-3 nS some activity be-
gan to spread further. It spread more slowly
than when fast synaptic inhibition was com-
pletely suppressed and not all cells partici-
pated (cf. Fig. 94.
The mechanism for inhibitory control over
the spread of population activity was exam-
ined as shown in Fig. 10. Simultaneous re-
1492 R. MILES, R. D. TRAUB, AND R. K. S. WONG
PTX t=Omin
*I
14 min
21 min
20 msec
B
gi = 36
2w ICI Jr-
9
A J-L
15
A -Al-
2CGGC
C gi
I
1
I I I
5 10 15
Block Number
OnS
2nS
3nS
1OnS
cells
FIG. 9. Spread of synchrony during partial block of inhibition. A: field potentials recorded during suppression of
inhibition by picrotoxin (100 PM). Electrodes were located at distances of 0.5, 3.5, and 6 mm from the stimulus. B:
numbers of cells firing in blocks 2,9, and 15 of the simulated array when gi, the conductance of unitary fast IPSPs,
was varied. A low level of cell firing spread throughout the array when gi was reduced to 3 nS. At 2 nS many more,
but not all, cells from each block fired. Propagation through the array was slow. Reducing gi to 0 nS resulted in firing
of all cells and increased the velocity of propagation. C: number of cells firing from each block from simulations with
gi values of 10, 3,2, and 0 nS.
cordings from cells at two sites were main- distant from the stimulus. Synaptic responses
tained as picrotoxin was added to suppress in- in cells at this distance normally possessed a
hibition (n = 8 slices). One site was l-3 mm prominent inhibitory phase and firing was
0 1
5
Distance mm
PTX t= Omin
-, -J
3-T
- w
mm A
15min
19min
100 msec
40mV
FIG. 10. Suppressing inhibition reveals latent EPSPs as synchrony begins to spread. Synaptic responses in cells 1
and 5 mm from the stimulus. In control solution (t = 0 min) an EPSP was followed by an IPSP in the cell at 1 mm.
No response was detected in the cell at 5 mm. At 15 min after adding 10 PM picrotoxin (PTX) a burst was evoked in
the cell at 1 mm and a slow EPSP was elicited with latency 40 ms in the distant cell. At 19 min synchronous bursts
were recorded in both cells.
SPREAD OF SYNCHRONY IN HIPPOCAMPUS 1493
rarely evoked. The other site was at least 5
mm away from the stimulus. No synaptic re-
sponse was detected over a range of mem-
brane potentials in cells at this distance. On
adding picrotoxin the IPSP in the cell closer
to the stimulus was suppressed and the EPSP
that was uncovered initiated a burst of several
action potentials (Fig. 10). At this time EPSPs
of slow time course were observed in the dis-
tant cell. Their latency was initially 35-50
ms. This conforms to the very slow spread of
population activity in simulations when inhi-
bition was reduced but not totally suppressed
(Fig. 9B). The latency of the EPSP in the dis-
tant cell then shortened and its amplitude
grew until firing was elicited and synchro-
nous bursting began to spread throughout the
slice. These results suggest that burst firing in
intercalated cells is needed to elicit EPSPs in
distant cells for synchrony to spread. Synap-
tic inhibition normally limits this process.
DISCUSSION
In the present study synchronous neuronal
firing was shown to spread throughout disin-
hibited longitudinal slices from the CA3 hip-
pocampal region (Fig. 2). We have focused on
synaptic mechanisms underlying the propa-
gation of synchrony. This seems justified
since the extent and speed of synchronous
burst propagation was correlated with a com-
ponent of synaptic events recorded in ele-
vated Ca (Fig. 5). Other factors may assist the
spread. Ephaptic interactions probably as-
sure tight synchronization of action poten-
tials within the propagating bursts (22, 54).
Electrotonic coupling exists in the hippocam-
pus (38) although it is not certain that all CA3
cells are coupled in this way. Changes of in-
terstitial Ca and K ions may be involved in
the genesis of some synchronous neuronal
population activities. However, these events
spread slowly at velocities in the range 0.00 l-
0.0 1 m/s (32, 34, 52). In contrast, synchro-
nous firing spread through longitudinal slices
at velocities in the range 0.12-o. 18 m/s (3 1).
Synaptic basis for the spread ofsynchrony
If chemical synaptic interactions are criti-
cal, what synaptic mechanism underlies the
spread of synchrony? We have attempted to
discriminate between two possible synaptic
A
20 /Jo po po
B
FIG. 11. A and B: 2 synaptic substrates for the spread
of synchrony. Excitatory connections are shown and S
represents fiber stimulation.
substrates which are illustrated in Fig. 11: I)
fiber stimulation excited axons which ran
throughout longitudinal slices (Fig. 11A).
Transmitter released from synapses formed
by these axons initiated synchronous bursts
and 2) alternatively axons were shorter than
longitudinal slices (Fig. 11B). Then syn-
chrony spread by a sequential process in
which active cells synaptically evoked firing
in distant cells which in turn generated EPSPs
to recruit more distant cells (1).
Our experimental and simulation studies
support the second scheme (Fig. 11B). First,
axons excited by stimulation did not appear
to extend throughout longitudinal slices (Fig.
3). Second, estimated axonal conduction ve-
locity was faster than the spread of synchrony
(Figs. 3 and 8). This scheme suggests that re-
current synaptic pathways between CA3 cells
mediate the longitudinal spread of synchrony
and that synaptic inhibition normally sup-
presses transmission in these recurrent path-
ways (9, 44). Simulations suggested that in-
hibitory control over the spread of synchrony
was graded rather than absolute (Fig. 9). The
uncovering of latent EPSPs as propagation
(Fig. 10) developed is consistent with this hy-
pothesis.
The scheme of Fig. 1lB also suggests that
fiber stimulation might sometimes elicit
EPSPs with two components. The first would
result from synapses of axons that were di-
rectly excited and the second from recurrent
synapses activated due to CA3 cell firing.
Such synaptic events were observed (Fig. 4)
when synaptic inhibition was blocked and ex-
1494 R. MILES, R. D. TRAUB, AND R. K. S. WONG
tracellular Ca was moderately elevated. EP-
SPs did not possess a second component in
10 mM Ca when neuronal threshold was con-
siderably increased (Fig. 3), as might be ex-
pected if the second component were depen-
dent on cell firing. We found the second
EPSP component was also blocked by paired
stimuli at an interval coincident with a post-
burst hyperpolarization that renders CA3
cells refractory and thus may prevent the gen-
eration of polysynaptic EPSPs (Fig. 4). EPSPs
with fast and slow components do not neces-
sarily involve the activation of two classes of
postsynaptic channels, with fast and slow ki-
netics, by transmitter released from a single
set of presynaptic terminals ( 15, 55).
Simulation studies
Synchronous discharges in disinhibited
transverse hippocampal slices spread at about
0.1 m/s compared to velocities of 0.3 to 0.5
m/s for conduction in mossy fibers or
Schaffer collaterals (4, 3 1, 5 1). One explana-
tion might be that synchrony is mediated via
axons which extend throughout the synchro-
nized cell population and which conduct es-
pecially slowly (58). However in longitudinal
slices the velocity with which synchrony
spread, 0.15 m/s, was slower than conduction
in longitudinal axons, 0.5 m/s. Further, ax-
ons excited by a point stimulus did not ex-
tend throughout these slices. Simulated neu-
ronal arrays were therefore constructed with
spatially limited excitatory connectivities. In
neuronal networks with this structure and
with many more neurons than previously
(58) synchrony spread more slowly than con-
duction in individual axons. The spatial pat-
tern of excitatory connections directly
affected the propagation velocity of synchro-
nized activity. The spread of synchrony ap-
proached axonal conduction velocity when
excitatory connections were not spatially lim-
ited (Fig. 7).
Anatomic considerations
Collateral synapses between CA3 pyrami-
dal cells mediate the spread of population ac-
tivity through longitudinal slices. These stud-
ies offer some insights into the organization
of these recurrent synaptic networks. First, it
is apparent that some collaterals run longitu-
dinally. Anatomic and physiological studies
have identified a longitudinal association
pathway in the CA3 region (6, 25, 36, 53).
Second, the amplitude of synaptic events was
progressively reduced as recordings were
made from cells at greater distances from a
fixed stimulus site. This could result if the
density of synaptic connections made by one
cell declined with distance from the cell-one
assumption made in constructing our simu-
lated neuronal network. Third, since synaptic
responses were not observed in cells > 4 mm
from a stimulus, pyramidal cell axon collater-
als appeared not to extend throughout these
slices. It must be recognized that some collat-
erals were probably cut during slice prepara-
tion. However dye injections made in vivo
have shown that CA3 pyramidal cells pos-
sess axon collaterals which arborize for dis-
tances > 1 mm but less than the longitudinal
extent of the hippocampus (16, 26). This is
further than the 400 pm extent estimated for
CA3 axon collaterals in transverse hippocam-
pal slices in experiments using focal gluta-
mate applications (9).
The rather uniform velocities at which syn-
chrony spread within each slice may offer fur-
ther insight into synaptic organization within
the CA3 region. In cortical regions with a co-
lumnar organization synchrony spreads at
velocities which fluctuate locally (11, 2 1).
These periodic fluctuations in velocity have
been attributed to spatial periodicities in hor-
izontal collateral arborizations of cortical py-
ramidal cells (18, 49). We did not examine
axon collateral morphology directly. How-
ever simulations yielded a uniform velocity
when the density of synaptic contacts made
by each cell was highest locally and declined
with longitudinal distance. Thus excitatory
synaptic networks in the CA3 region may be
organized according to a rather simple pat-
tern (47). This pattern does not seem to be
constrained by the transverse lamellar orga-
nization proposed for the hippocampus (3).
ACKNOWLEDGMENTS
We thank Dr. Angelo Rossi for assistance in using the
IBM 3090 vector facility.
This work was supported by grants from the National
Institutes of Health and the Klingenstein Foundation.
Received 13 January 1988; accepted in final form 19
May 1988.
SPREAD OF SYNCHRONY IN HIPPOCAMPUS 1495
REFERENCES
1.
2.
3.
4.
5 . .
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
18.
ADRIAN, E. D. The spread of activity in the cerebral
cortex. J. Physiol. Lond. 88: 127- 16 1, 1938.
ALGER, B. E. AND NICOLL, R. A. Spontaneous in-
hibitory post-synaptic potentials in the hippocam-
pus: mechanism for tonic inhibition. Brain Res. 200:
195-200, 1980.
ANDERSEN, P., BLISS, T. V. P., AND SKREDE, K. K.
Lamellar organization of hippocampal excitatory
pathways. Exp. Brain Res. 13: 222-238, 197 1.
ANDERSEN, P., SILFVENIUS, H., SUNDBERG, S. H.,
SVEEN, O., AND WIGSTROM, H. Functional charac-
teristics of unmyelinated fibres in the hippocampal
cortex. Brain Res. 144: 1 l- 18, 1978.
AYALA, G. F., DICHTER, M., GUMNIT, R. J., MAT-
SUMOTO, H., AND SPENCER, W. A. Genesis of epi-
leptic interictal spikes. New knowledge of cortical
feedback systems suggests a neurophysiological ex-
planation of brief paroxysms. Brain Res. 52: l- 17,
1973.
BARTESAGHI, R., GESSI, T., AND SPERTI, L. Interla-
mellar transfer of impulses in the hippocampal for-
mation. Exp. Neural. 82: 550-567, 1983.
BERRY, M. S. AND PENTREATH, V. W. Criteria for
distinguishing between monosynaptic and polysyn-
aptic transmission. Brain Res. 105: l-20, 1976.
BROWN, T. H. AND JOHNSTON, D. Voltage-clamp
analysis of mossy fiber synaptic input to hippocam-
pal neurons. J. Neurophysiol. 50: 487-507, 1983.
CHRISTIAN, E. P. AND DUDEK, F. E. Characteristics
of local excitatory circuits studied with glutamate
microapplication in the CA3 area of rat hippocam-
pal slices. J. Neurophysiol. 59: 90- 109, 1988.
COLLINGRIDGE, G. L., GAGE, P. W., AND ROBERT-
SON, B. Inhibitory post-synaptic currents in rat hip-
pocampal neurones. J. Physiol. Lond. 356: 55 l-564,
1984.
CONNORS, B. W. AND CHERVIN, R. D. Physiological
evidence for periodicity and directionality of lateral
excitatory connections in rat neocortex. Sot. Neu-
rosci. Ahstr. 12: 350, 1986.
CROWELL, R. M. Distant effects of an epileptogenic
focus. Brain Res. 18: 137-l 54, 1970.
DICHTER, M. AND SPENCER, W. A. Penicillin-in-
duced interictal discharges from the cat hippocam-
pus. I. Characteristics and topographical features. J.
Neurophysiol. 32: 649-662, 1969.
DICHTER, M. AND AYALA, G. F. Cellular mecha-
nisms of epilepsy: a status report. Science Wash. DC
237: 157-164, 1987.
DINGLEDINE, R., HYNES, M. A., AND KING, G. L.
Involvement of n-methyl-d-aspartate receptors in
epileptiform bursting in the rat hippocampal slice. J.
Physiol. Lond. 300: 175-l 89, 1986.
FINCH, D. M., NOWLIN, N. L., AND BABB, T. L.
Demonstration of axonal projections of neurons in
the rat hippocampus and subiculum by intracellular
injection of HRP. Brain Res. 27 1: 20 1-2 16, 1983.
FRANKENHAEUSER, B. AND HODGKIN, A. L. The
action of calcium on the electrical properties of
squid axons. J. Physiol. Land. 137: 2 18-244, 1957.
GILBERT, C. D. AND WIESEL, T. N. Clustered intrin-
sic connections in cat visual cortex. J. Neurosci. 3:
1116-l 133, 1983.
19. GINSBORG, B. L. Electrical changes in the mem-
brane in junctional transmission. Biochim. Biophys.
Acta 300: 289-3 17,1973.
20. GOLDENSOHN, E. S. AND SALAZAR, A. M. Temporal
and spatial distribution of intracellular potentials
during generation and spread of epileptogenic dis-
charges. In: Basic Mechanisms of the Epilepsies, Ad-
vances in Neurology, edited by A. V. Delgado-Es-
cueta, A. A. Ward, D. M. Woodbury, and R. J. Por-
ter. New York: Raven, 1986, vol. 44, p. 559-582.
2 1. GUTNICK, M. J. AND WADMAN, W. J. Intrinsic neu-
22.
23.
24.
25.
26.
27.
28.
29.
30.
31.
32.
33.
34. LEAO, A. A. P. Spreading depression of activity in
ronal connectivity in neocortical brain slices as re-
vealed by non-uniform propagation of paroxysmal
discharges. Sot. Neurosci. Ahstr. 12: 349, 1986.
HAAS, H. L. AND JEFFERYS, J. G. R. Low-calcium
field burst discharges of CA 1 neurones in rat hippo-
campal slices. J. Physiol. Lond. 354: 185-20 1, 1984.
HABLITZ, J. J. Picrotoxin induced epileptiform ac-
tivity in hippocampus: role of endogenous versus
synaptic factors. J. Neurophysiol. 5 1: 10 1 l- 1027,
1984.
HILLE, B. Charges and potentials at the nerve sur-
face: divalent cations and pH. J. Gen. Physiol. 5 1:
221-236,1968.
HJORTH-SIMONSEN, A. Some intrinsic connections
of the hippocampus in the rat: an experimental anal-
ysis. J. Comp. Neural. 147: 145- 162, 1973.
ISHIZUKA, N., KRZEMIENIEWSKA, K., AND
AMARAL, D. G. Organization of pyramidal cell axon
collaterals in field CA3 of the rat hippocampus. Sot.
Neurosci. Abstr. 12: 1254, 1986.
JAHR, C. E. AND JESSELL, T. M. Synaptic transmis-
sion between dorsal root ganglion and dorsal horn
neurons in culture: antagonism of monosynaptic ex-
citatory post-synaptic potentials and glutamate exci-
tation by kynurenate. J. Neurosci. 5: 228 l-2289,
1985.
JEFFERYS, J. G. R. Influence of electric fields on the
excitability of granule cells in guinea pig hippocam-
pal slices. J. Physiol. Land. 300: 448-460, 198 1.
JOHNSTON, D. AND BROWN, T. H. Giant synaptic
potential hypothesis for epileptiform activity. Sci-
ence Wash. DC 2 11: 294-297, 198 1.
KNOWLES, W. D. AND SCHWARTZKROIN, P. A. Lo-
cal circuit synaptic interactions in hippocampal
brain slices. J. Neurosci. 1: 3 18-322, 198 1.
KNOWLES, W. D., TRAUB, R. D., AND STROW-
BRIDGE, B. W. The initiation and spread of epilepti-
form bursts in the in vitro hippocampal slice. Neuro-
science21: 441-455, 1987.
KONNERTH, A., HEINEMANN, U., AND YAARI, Y.
Nonsynaptic epileptogenesis in the mammalian hip-
pocampus. I. Development of seizurelike activity in
low extracellular calcium. J. Neurophysiol. 56: 409-
423, 1986.
LACAILLE, J.-C., MUELLER, A. L., KUNKEL, D. D.,
AND SCHWARTZKROIN, P. A. Local circuit interac-
tions between oriens-alveus interneurons and CA 1
pyramidal cells in hippocampal slices: electrophysi-
ology and morphology. J. Neurosci. 7: 1979-1993,
1987.
1496 R. MILES, R. D. TRAUB, AND R. K. S. WONG
35.
36.
37.
38.
39.
40.
41.
42.
43.
44.
45.
46.
47.
48.
the cerebral cortex. J. Neurophysiol. 7: 359-390,
1944.
LEBOVITZ, R. M., DICHTER, M., AND SPENCER,
W. A. Recurrent excitation in the CA3 region of cat
hippocampus. ht. J. Neurosci. 2: 99- 108, 197 1.
L~RENTE DE No, R. Studies on the structure of the
cerebral cortex. II. Continuation of the study of the
ammonic system. J. fir Psychol. Neural. Leipzig 46:
113-177,1934.
MACVICAR, B. A. AND DUDEK, F. E. Local synaptic
circuits in rat hippocampus: interactions between
pyramidal cells. Brain Res. 184: 220-223, 1980.
MACVICAR, B. A. AND DUDEK, F. E. Electrical cou-
pling between pyramidal cells: a direct demonstra-
tion in rat hippocampal slices. Science Wash. DC
213: 782-785,1981.
MILES, R. AND WONG, R. K. S. Single neurones can
initiate synchronized population discharge in the
hippocampus. Nature Lond. 306: 37 l-373, 1983.
MILES, R. AND WONG, R. K. S. Unitary inhibitory
synaptic potentials in the guinea-pig hippocampus
in vitro. J. Physiol. Lond. 356: 97-l 13, 1984.
MILES, R., WONG, R. K. S., AND TRAUB, R. D. Syn-
chronized afterdischarges in the hippocampus: con-
tribution of local synaptic interactions. Neurosci-
ence 12: 1179-l 189,1984.
MILES, R. AND WONG, R. K. S. Excitatory synaptic
interactions between CA3 neurones in the guinea-
pig hippocampus. J. Physiol. Lond. 373: 397-418,
1986.
MILES, R. AND WONG, R. K. S. Inhibitory control
of local excitatory circuits in the guinea pig hippo-
campus. J. Physiol. Lond. 388: 6 1 l-629, 1987.
MILES, R. AND WONG, R. K. S. Latent synaptic
pathways revealed after tetanic stimulation in the
hippocampus. Nature Lond. 329: 724-726, 1987.
MISGELD, U. AND FROTSCHER, M. Post-synaptic
GABA-ergic inhibition of non-pyramidal cells in the
guinea-pig hippocampus. Neuroscience 19: 193-
206,1986.
NEWBERRY, N. R. AND NICOLL, R. A. Comparison
of the action of baclofen with -amino-butyric acid on
rat hippocampal pyramidal cells in vitro. J. Physiol.
Lond. 360: 161-185, 1985.
NJA, A. AND P~RVES, D. Specific innervation of
guinea-pig superior cervical ganglion cells by pre-
ganglionic fibres arising from different levels of the
spinal cord. J. Physiol. Lond. 264: 565-583, 1977.
PENFIELD, W. AND JASPER, H. Epilepsy and the
Functional Anatomy qf the Human Brain. Boston,
MA: Little, Brown, 1954.
49.
50.
51.
52.
53.
54.
55.
56.
57.
58.
59.
60.
61.
ROCKLAND, K. S. AND LUND, J. S. Widespread peri-
odic intrinsic connections in the tree shrew visual
cortex. Science Wash. DC 2 15: 1532- 1534, 1982.
RUTECIU, P. A., LEBEDA, F. J., AND JOHNSTON, D.
Epileptiform activity induced by changes in extra-
cellular potassium in hippocampus. J. Neurophysiol.
54: 1363-1371, 1985.
SCHWARTZKROIN, P. A. AND PRINCE, D. A. Cellular
and field potential properties of epileptogenic hippo-
campal slices. Brain Res. 147: 117- 130, 1978.
SNOW, R. W., TAYLOR, C. P., AND DUDEK, F. E.
Electrophysiological and optical changes in slices of
rat hippocampus during spreading depression. J.
Neurophysiol. 50: 56 l-572, 1983.
SWANSON, L. W., WYSS, J. M., AND COWAN, W. M.
An autoradiographic study of the organization of in-
trahippocampal association pathways of the rat. J.
Comp. Neural. 181: 681-716, 1978.
TAYLOR, C. P. AND DUDEK, F. E. Synchronous neu-
ral afterdischarges in rat hippocampal slices without
active chemical synapses. Science Wash. DC 2 18:
810-812,1982.
THOMSON, A. M. AND WEST, D. C. N-methylaspar-
tate receptors mediate epileptiform activity evoked
in some but not all conditions in rat neocortical
slices. Neuroscience 19: 116 1- 1177, 1986.
TRAUB, R. D. Simulation of intrinsic bursting in
CA3 hippocampal neurons. Neuroscience 7: 1233-
1242,1982.
TRAUB, R. D. AND WONG, R. K. S. Cellular mecha-
nism of neuronal synchronization in epilepsy. Sci-
ence Wash. DC 2 16: 745-747, 1982.
TRAUB, R. D., KNOWLES, W. D., MILES, R., AND
WONG, R. K. S. Models of the cellular mechanism
underlying propagation of epileptiform activity in
the CA2-CA3 region of the hippocampal slice. Neu-
roscience 2 1: 457-470, 1987.
TRAUB, R. D., MILES, R., AND WONG, R. K. S.
Models of synchronized hippocampal bursts in the
presence of inhibition. 1. Single population events.
J. Neurophysiol. 58: 739-75 1, 1987.
WILDER, B. J. Experimental studies, models and
phylogenetic aspects of secondary epileptogenesis.
In: Secondary Epileptogenesis, edited by A. Mayers-
dorfand R. P. Schmidt. New York: Raven, 1982, p.
27-43.
WONG, R. K. S. AND TRAUB, R. D. Synchronized
burst discharge in the disinhibited hippocampal
slice. I. Initiation in the CA2-CA3 region. J. Neuro-
physiol. 49: 442-458, 1983.