281

Electroencephalography and Clinical Neurophysiology, 1975, 38:281-293 "{3 Elsevier Scientific Publishing Company, Amsterdam - Printed in The Netherlands

INTRACORTICAL

TRANSMISSION

ACROSS THE CEREBRAL

OF SPECIFIC EVOKED

ACTIVITY

CORTEX 1

J. A. ENNEVER 2 Department of Physiology, University College, London W.C.1 (Great Britain} (Accepted for publication: October 14, 1974)

There appear to be three main types of purely intracortical tangential propagation of activity described in the literature on the cortex. One type is characterized by a very slow speed of conduction, 0.001 m/sec-0.001 m/min, and includes the Jacksonian march (Jackson 1931), possibly the movement of ophthalmic migraine scotomas (Milner 1958), and spreading depression. Le~o first described the latter: a wave of intense activity spreads with a velocity of about 0.005 m/rain, in a gradually expandin~ circle across the cortex, leaving the tissue electrically inactive for about 1 min afterwards (Le~o 1944). Burns (1958) described a similar slow-spreading phenomenon in slabs of isolated cortex, which supports the idea that this type of spread is a purely intracortical phenomenon. The transmission is thought to involve synaptic excitation, facilitated by the accumulation of extracellular potassium ions (Grafstein 1956; Vysko~il et al. 1972), and is triggered by the simultaneous activation of an abnormally large number of cells. Thus, this first type of spreading activity is highly abnormal. The second, and faster, type of transcortical transmission travels at a velocity of 0.1-4).2 m/sec in a network of cells 1 mm or more beneath the surface. Burns (1951, 1958) evoked this type of spread in chronic isolated slabs of unanaesthetized cortex by a weak direct stimulus, which was not strong enough to produce spreading depression. This "burst response" spread unattenuated throughout the 20 mm long slab, and Burns This research was supported by a Medical Research Council Training Grat~t. 2 Present address: Brain Research Institute, University of Zurich, August-Forel-Strasse 1, CH-8029 Zurich.

concluded that this demonstrated a potential for unlimited-synaptic-tangential transmission inside the normal cortex. Another example of this type of spreading activity was described by Cobb et al. (1955) in isolated cortex, but in the presence of anaesthetic. In this case the spreading "activity" was a strychnine spike, which does not usually "spread" intracortically more than about 2 mm (Dusser de Barenne and McCulloch 1938, and personal observation), but Cobb et al. induced the intracortical transmission by strychninization of the cortex at 5-7 mm intervals. The same experiments in intact cortex were complicated by a much faster spread, via corticocortical fibres. However, only intracortical, not intercortical, transmission is being considered here. Also included in the second group, should be the apparent spread of activity evoked by weak repetitive direct stimulation of the cortex (Adrian (1936) in anaesthetized brain; Penfield and Boldrey (1937) in unanaesthetized patients; Burns and Smith (1962) in unanaesthetized cat cortex). The exact pathway for these spreading influences is not known, but it seems likely that the repetitive stimulation increases the excitability of a deep cortical network in a paroxysmal fashion, and gradually induces the second type of transmission. The third, and fastest, type of intracortical transmission is that evoked by a single weak direct stimulus to the cortex, again a highly abnormal stimulus. This evokes a surface-negative wave which travels up to 4-5 mm from the stimulus site in layer I (Chang 1950; Ochs and Suzuki 1965). Burns (1958) detected the response up to 10 mm away. The velocity was estimated as 1 m/sec in cat and 0.6--0.7 m/sec in monkey by

282

C h a n g (1950) a n d 2 m/sec in cat by Burns (1958). T h e p r o p e r t i e s o f these three types of s p r e a d i m p l y t h a t there a r e p o t e n t i a l t r a n s c o r t i c a l p a t h ways from a n y one cortical p o i n t to a n y other. H o w e v e r , it is i m p o r t a n t to notice that all these types of s p r e a d have only been seen as the result of highly a b n o r m a l cortical stimulation. H o w these p r o p e r t i e s of cortical circuitry are used in the t r a n s m i s s i o n of n o r m a l cortical activity is not known. T h e e x p e r i m e n t s d e s c r i b e d here were designed to investigate w h e t h e r the short latency r e s p o n s e e v o k e d by w e a k s o m a t i c stimulation, which was c o n s i d e r e d to be a relatively n o r m a l response, i n v o l v e d a n y o f the three types of i n t r a c o r t i c a l s p r e a d d e s c r i b e d above.

J.A. ENNEVER

METHODS B l a c k h o o d e d rats of the Lister strain a n d albin o W i s t a r rats were used. O n l y the y o u n g e r rats, 170-200 g, were used when a n extensive craniot o m y was p e r f o r m e d , since the fibrous connections between t h e d u r a a n d c r a n i u m were so t o u g h on o l d e r rats that the r e m o v a l o f a large a r e a o f b o n e caused s u b d u r a l h a e m o r r h a g e s . T h e rats were a n a e s t h e t i z e d with 1.6-1.9 g / k g b o d y weight of urethane, a d m i n i s t e r e d i.p. as 3 6 ~ solution. U r e t h a n e ( e t h y l - c a r b a m a t e ) was used since it p r o d u c e s a s t e a d y d e p t h o f a n a e s thesia for 6-10 h ; a n d secondly, B i n d m a n et al. (1964a) have s h o w n that u n d e r u r e t h a n e the

4 J

2

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Fig. 1. Topographical maps of the cerebral hemisphere of the rat. A : Right side: localization of sensory and motor function in the rat (Woolsey 1958). Left side: cytoarchitectonic map of the cortex (Krieg 1963). Dotted lines represent the sutures of the skull which are used for landmarks in the rat, in the absence of sulci. B: Drawn to same scale (each division = ! mm). Dotted lifie: average boundary of the cortical somatosensory receiving area of the contralateral forepaw (CRA), in urethane anaesthetized rats. Shaded region: central area. Central dot: position at which the largest responses were recorded. Boundary responses were usually recorded from the posterior boundary of the CRA, just inside or on the dotted line.

283

INTRACORTICAL TRANSMISSION OF EVOKED POTENTIALS

potential level of the cortex reflected the state of excitation of the subjacent cells. Thus it was convenient to use the potential level of the cortex as an estimate of cortical activity. A trephine hole, 4 mm in diameter, was made over the forepaw receiving area (Fig. 1), the dura was removed and a small ring cemented with dental wax round the hole. This enabled the cortex to be covered with liquid paraffin. The rectal temperature was maintained at 36-37 ° C through a transistor controlled heating device. The sensory stimulus used was a weak electrical pulse, 0.05 msec duration, delivered to the palmar skin of the contralateral forepaw, through a pair of fine needles. The stimulus strength was always subthreshold for movement. The surface records were taken from the cortex with 0.5 mm silver-ball electrodes, coated with AgC1, or glass-pipettes, 4-8 #m tip diameter, filled with 10 % NaC1. The latter were also used for monitoring multi-unit activity during polarisation. The indifferent electrode was placed on the eye. The recording electrodes were connected to Tektronix 3 A 3 amplifiers in the direct-coupled mode, or with a 300 msec time constant, via cathode followers. Measurements were taken from a continuous film of the oscilloscope display. To change the excitatory state of various elements in the cortex a piece of filter paper 2 mm square soaked in 2 % strychnine was applied directly to the cortex for 1 min ; or 2.5 % solutions of GABA were placed on the cortex and washed away after 10 min. Intravenous injection of 0.3 ml of 2 % strychnine solution, which usually produced convulsions, was also used. A fourth method of artificially altering the excitation state was by polarization. Voltages, connected in parallel with a 100 pF capacitor, could be applied between a saline soaked cotton wick on the cortical surface and a second electrode in the mouth, through a 2 Mf~ potentiometer. Currents of 50-300 pA/mm 2 were used. Assuming a linear functional relationship between the electrocorticograms (ECoGs) at two different recording sites, the cross-correlation coefficient, rxy (T) (where the mutual displacement in time T was always zero, i.e., zero timelag) was estimated in order to measure the degree of synchrony of the two signals x (t) (electrode 1) and Yi (t) (successive electrodes 2, 3, 4, 5

or 6, (i), more posterior). The correlation coefficient was calculated with a Packard Bell 440 from the cross-variance (lagged product-moment correlation), with zero mean ; Cxy(T) = E xt,)" yr, + T)

-

-

X~n)" ~ Ytn + T) N

where n is the sample index at time t; 0 < t < 30 secs; 1 < n < N, the sample rate was 2500/sec, thus N = 2500 x 30; and normalized by the autocovariance functions : Cxy rx,= x / C x x . Cyy giving:

=(

r~y

n=l

x ~o,.y,o, - [ ~n 1 x ,°,. n=Es yi,n,] fi1

N q2~l~( N I- N 72~_~ / I1=1 Z x:,-[l- n=lEx,.,J _J()Y~ ~n=l y,~.,-L._E Yi ,o,J

The six silver ball electrodes were spaced at 1 mm intervals in a line across the cortex, in an anteriorposterior orientation. The simultaneous ECoGs, recorded between each electrode and ground were fed into a digital data acquisition system, which converted the signal amplitude to 6 bit sample values (bandwidth 10,000-12 c/sec). Since the apparatus could only read 4 E C o G channels simultaneously, the data for Fig. 6 were recorded in two sets of four (i.e., electrodes 1, 2, 3, 4; then electrodes 1, 4, 5, 6). Each correlation coefficient was calculated from a 30 sec E C o G sample and averaged with the values obtained in 3 immediately subsequent trials. RESULTS

Under most anaesthetics, short latency responses evoked by a shock to the contralateral forepaw can be recorded over about half of the hemisphere in the rat. This is called here the cortical forepaw area " C R A " (see area enclosed by the dotted line in Fig. 1, B). The hatched area, in Fig. 1, B encloses the central region of the CRA where the responses, plotted under urethane anesthesia, are greater than half maximum amplitude; this is the region usually referred to as the primary sensory receiving area of the

284 forepaw in the rat (Libouban-Letouz6 1963: Ennever 1971). However, outside this central region smaller short latency responses can still be recorded• The smallest responses, on the boundary of the CRA (marked by the dotted line in Fig. 1, B), will be referred to as boundary responses. Boundary responses are not the result of electrotonic spread from the central area (Ennever 1971). They are accompanied by spike activity in the subjacent cortex, and are not abolished by cuts entering layer VI of the cortex, which would block intracortical transmission from the centre of the CRA. Theretore boundary responses are not themselves simply a result of intracortical spread; they are independent short latency responses, and are used here to study intracortical transmission. Evidence for the presence of intracortical transmission of evoked potentials would be demonstrated if purely cortical changes could influence the position of the CRA boundary, i.e., induce responses in a region previously outside the CRA or reduce the size of the CRA. No such evidence, however, could be found in the attempts to do this described below• The estimation of spread in terms of mass responses in the centre of the CRA is considered in the second half of the Results section. 1. Boundary responses In an attempt to induce or enhance intracortical spread of the evoked responses, or some component of them, the posterior boundary regions of the CRA (those regions of the cortex on the posterior portion of the dotted line in Fig. 1, B) were treated with strychnine. This appears to increase the excitability of some cortical elements that are activated by the afferent volley. Topical application to about 2 mm 2 produced spontaneous strychnine spikes in an area of 3-4 mm 2 local to the application; but the shape of the boundary evoked responses was unchanged (Fig. 2) and no change in the location of the CRA border was ever found. A second method of locally increasing the activity of some cortical elements in order to induce or just alter the intracortical transmission of primary evoked activity, was to pass a small, steady current (50-500/zA/mm 2) across the cortex (polarization). Purpura and McMurtry (1965)

J.A. ENNEVER

TOPICAL APPLICATION OF STRYCHNINE TO POSITION 4. before

after

"?00LLV '30 msec

1234 Fig. 2. Responses evoked by stimulation of the contralateral forepaw, recorded at 1 mm intervals simultaneously from the surface of the cerebral cortex [diagrammatically illustrated below). Electrodes I and 2 are in the central CRA. electrodes 3 and 4 in the more peripheral region. Point 4 is on the CRA boundary. A 2 mm 2 piece of filter paper soaked in 2 o~, strychnine was placed for 1 min on the CRA boundary; spontaneous strychnine spikes occurred at point 4. and could also be detected at 3. No detectable difference could be found in the boundary evoked responses at this point: the relative pattern of responses was unchanged. On this and all other figures• upward deflections signal positivity of the focal electrode.

have shown that surface-positive current (SP) tends to increase the spontaneous firing of some neurones deeper than 0.7 mm4).9 mm in the cortex, whereas surface-negative current (SN) inhibits it• The current has the opposite effect on more superficial cells• The standard changes in the shape of the mass response were seen: SP increased negative components, SN increased positive components (Bindman et al. 1964b). (However, it was noticed that these changes were reversed in the mass responses recorded 3 mm away from the polarizing wick,

INTRACORTICALTRANSMISSIONOF EVOKED POTENTIALS

285

EFFECTS OF CORTICAL POLARIZATION

fire spontaneously; these are interspersed with periods when m a n y cells can be found to fire ON EVOKED RESPONSES. spontaneously. The active bursts are accompanied by a positive D C shift of the surface pobefore during tential (see Fig. 7). These periods of activity are 500lay also related to an increased state of arousal, since any alerting stimulus will evoke such a burst, SN and they occur more frequently and eventually 10reset 5oopv become continuous as the anaesthetic depth is reduced. However, it was found that these bursts SP of generalized cortical activity do not affect the 10msec 5oopv position of the boundary of the CRA. Neither did sub-convulsant and supra-convulsant doses SP of strychnine which also affect the central excita10msec tory state in some way (Fig. 5) (Smolin and Samko Fig. 3. 1: Effect of surface-negative (SN) polarization of 50 1968). The convulsant did increase the amplitude #A/mm2 on cortical responses evoked by contralateral fore- of the primary potentials, but as with polarization paw stimulation. Potentials are recorded from the centre of the change in amplitude was proportional to the CRA. Notice typicalincrease in surface-positivecomponents. original size of the response, and no change in 2: Effect of surface-positive(SP) current of 200/~A/mm2 on a response recorded more peripheral in the CRA than 1. the position of the CRA boundary was found in Note typical increase in surface-negative components. any experiment. 3: Boundary response: 200/zA/mm2 SP increases a negative Thus none of the direct or induced manipulacomponent but the change is very small, proportional to the tions of cortical activity had any effect on the original amplitude.

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presumably because the direction of the effective current is reversed in that part of the field.) A microelectrode monitoring the activity of deep neurones showed that even when deeper structures were facilitated or inhibited by the current (which was far greater than the current intensities that maximally affect the surface mass response), no change in the boundary of the CRA could be detected, and no component of the evoked responses could be induced beyond the original boundary. The current only changed the shape of the existing evoked potentials in the usual manner (Fig. 3). The amplitude of the change produced by a supramaximal current was found to be directly proportional to the original peak-to-peak amplitude (see Fig. 4) and extrapolation of the graph to zero confirms the observation that polarization could not induce a response where previously none existed (i.e., peak-to-peak a m p l i t u d e = 0 ) . This is not the effect that would be expected if the afferent volley evoked activity in an unlimited network, whose excitability was increased by the polarizing current. Under urethane anaesthesia, there are quiescent periods in the cortex during which n o cells

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O.Ol 0.1 .o IO Peak-to-peak amplitudeof evokedpotentials (my) Fig. 4. Notice direct proportionality ofchange in peak-to-peak amplitude of the cortical evoked responses, plotted on a log-log scale, with the amplitude of the response before passing the polarizingcurrent. The potentials were evoked by electrical stimulation of the contralateral forepaw.The results shown on the graph include changes induced by surfacepositive (SP) and surface-negative (SN) current (current densities of 50-300 gA/mm2, and durations of 2-60 min). The direct relationship revealed by the graph is therefore independent of the dimensions of these large currents.

286

J.A. ENNEVER

l o c a t i o n o f the e v o k e d potential. It s h o u l d be e m p h a s i z e d t h a t a decrease in C R A w o u l d also have p r o v i d e d positive evidence for the existence of lateral t r a n s m i s s i o n of the e v o k e d response. H o w e v e r , the results all i m p l i e d t h a t the b o u n daries o f the f o r e p a w a r e a at a n y one time a r e rigid. It is p o s s i b l e t h a t the r i g i d i t y is the result of a n a t o m i c a l limitations, since a t t e m p t s a t altering the p h y s i o l o g i c a l state o f the cortex d i d n o t induce a n y lateral t r a n s m i s s i o n o f the response. Very o c c a s i o n a l l y a n e v o k e d negative wave w o u l d a p p e a r successively a l o n g a r o w o f electrodes, even a l o n g t h o s e o u t s i d e the C R A b o u n dary. H o w e v e r , n e i t h e r their a m p l i t u d e o r freq u e n c y was c h a n g e d b y the a t t e m p t s to alter cortical excitability.

EEG Electrodes lmm

apart

2 mm apart

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OF STRYCHNINE

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5 mm

Fig. 6. The ECoG recorded from points l mm apart yield very similar traces lupper set of records, retouched). If the electrodes are moved 2 mm apart differences marked by arrows become obvious. Graph: data from experiments in which 6 recording electrodes were used. Each point is a quantitative estimation of the variation between two simultaneous ECoG records. The correlation coefficient is a measure of the synchrony between activity from electrodes placed 1-5 mm apart. The graph shows the relative changes over 5 mm.

|

i•

i

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30 msec

1234

Fig. 5. Responses evoked by stimulation of the contralateral forepaw recorded at 1 mm intervals, simultaneously from the surface of the cerebral cortex. After a convulsant dose of strychnine (30. mg/kgl, the amplitude of the central responses, 1 and 2, increased due to a change in the excitation state of the brain: however, the peripheral responses. 3 and 4. were not significantly enhanced.

2. Central responses E v o k e d responses r e c o r d e d at two p o i n t s near the centre o f the C R A . a b o u t 2 m m a p a r t , are usually similar in c o n f i g u r a t i o n , a n d their s h a p e varies in a n identical m a n n e r . T h e variations in s h a p e have been s t u d i e d by B i n d m a n et al. (1964a), w h o have shown t h a t the shape of a r e s p o n s e varies with the s p o n t a n e o u s activity in the c o r t e x o n the a r r i v a l o f the afferent volley. T h e p e a k - t o - p e a k a m p l i t u d e o f the responses a r e r o u g h l y p r o p o r t i o n a l to the a m p l i t u d e o f the surface-positive D C shift which a c c o m p a n i e s the s p o n t a n e o u s activity (i.e.. E C o G a m p l i t u d e ) . E x p e r i m e n t s were designed to d i s c o v e r w h e t h e r the changes in s h a p e of the t w o s i m u l t a n e o u s l y r e c o r d e d responses were closely c o u p l e d because of i n t r a c o r t i c a l spread, o r the result o f identical fluctuations in the cortical activity at each point.

INTRACORTICALTRANSMISSIONOF EVOKED POTENTIALS

a. The instantaneous synchrony of the cortical activity at two points E E G activity was recorded simultaneously from six surface electrodes, placed in a line at 1 m m intervals across one hemisphere, and with n o specific relationship to the CRA. The sync h r o n y between the E C o G at the first electrode and each of the other five was measured a n d expressed in terms of a correlation coefficient. The results (Fig. 6, graph) show that there is a high correlation between the instantaneous activity at the two points, which decreases with increasing distance over a range of a b o u t 2 m m before reaching a constant level. Therefore, it is possible that the closely coupled variations in the shape o f the evoked responses were due only to the high correlation of the s p o n t a n e o u s activity at each point.

b. Shape of two simultaneously recorded responses when cortical excitation states at the two recording sites differ W h e n a burst of s p o n t a n e o u s activity occurred

287

at one point a n d n o t the other, and it was arranged for a stimulus to be delivered at such an instant, the shape of the responses was completely different. E a c h evoked response always assumed the appropriate s h a p e for the level o f s p o n t a n e o u s activity at its recording site (Fig. 7). T h u s the Shape of the p r i m a r y evoked response is mainly determined by the state of a local area o f cortex less than a b o u t 2 m m in radius, a n d it is not strongly affected by activity 2 m m or m o r e away.

c. Measurement of the dependence of evoked response on remote cortical excitation A l t h o u g h the responses 2.5 m m apart (Fig. 7) appeared to behave independently, it was possible that the burst o f s p o n t a n e o u s activity at one point still had a small influence on the evoked activity in a neighbouring area. Therefore, the peak-topeak amplitude (a) of between 30-90 responses, at one point, was correlated with the coincident levels of s p o n t a n e o u s cortical activity at points 1.5-2:5 m m away. In fact, since the level of s p o n t a n e o u s activity is directly p r o p o r t i o n a l to

I0 m s e c Fig. 7. Two top traces: ECoG recorded at two points 2.5 mm apart in the. central CRA. I, II and III are taken from the same records at times indicated by arrow, but on an expanded time scale. They show the responses evoked by weak electrical stimulation of the contralateral forepaw, more clearly. The responses in III are typical of evoked potentials taken when the cortex is "quiescent" (DC level=0). This stimulus also generates a burst of activity after a cortical response (see top 2 traces). Responses in II are entirely negative, both were evoked during a burst of activity, when the potential level of the cortex was positive with respect to the baseline. I shows the positive and negative forms occurring together. It can he seen from I that there is a burst of activity (DC level positive) at one point and not at the other, and the evoked potentials at this instant vary completely independently of each other. Voltage calibration = 1 mV.

288

J.A. ENNEVER

the DC level of the cortex (Bindman et al. 1964a), in this preparation the DC level (B) was used in the correlations as the measure of the level of spontaneous activity. It was shown above that there is a high degree of correlation between the ongoing spontaneous activity at two adjacent points: therefore since (a) is very highly correlated with the DC level (A) at that point, (a) will also be highly correlated with (B), the DC level 1 or 2 mm away.However, the aim of the present calculation was to try to uncover some relationship between (B) and [a) independent of this correlation. To do this the following analysis was performed. The correlation coefficient ra.A for (a) on (A) was calculated, and similarly the coefficients ra.B and rA. B" Using these three correlation coefficients, the partial correlation coefficients were calculated for the dependence of (a) on (A) when the variations due to (B) are excluded, i.e., when (B) is effectively kept constant (r~A.B), and (a) on (B) when (A) is effectively kept constant (r~B.A). The significance of these coefficients was found from standard statistics tables. The results of eight analyses are shown in Table I. In none of the cases where (A) and (B) vary significantly independently (i.e., raA,B has P