‘Physiolony and Behavior, Vol 10, pp. 1005-1008. in Research Publications Inc, 1973, Printed inthe US A. Correlation of Evoked Potentials in the Caudate Nucleus and Conditioned Motor Responses’ J. GRINBERG-ZYLBERBAUM?, R. PRADO-ALCALA AND H. BRUST-CARMONA Physiology Department, Faculty of Medicine and Psychology School, ‘National University of Mexico, Mexico 20, D. F. (Received 10 October 1972) GRINBERG-ZYLDERBAUM, J., R. PRADO-ALCALA AND H. BRUST-CARMONA. Correlation of evoked potent in the caudete nucleus end conditioned motor responses. PHYSIOL. BEHAV. 10(6) 10051009, 1973 The relevant pattcpation of the caudate mucleus (CN) in motor conditioned responses has been described mn diferent papers. Tht function could be reflected by changes init electrical activity. In cats with implanted electrodes in the CN the evoked potentials produced by the conditioned stimulus (light or sound) appeared and increased duting the performance of an "pproaching conditioned response (CR). Extinction ofthe CR caused decrease in amplitode while reconditioning had the ‘opposite effect. The potentials did not decrease as long 36 the conditioning sessions were repeated. The correct performance of the CR was postively cortelated at the level of <0.001 with the amplitude or te appearance of the tvoked potentals. The increment of the evoked potentials seems to be independent of the cat's movements since the ‘movements were much more (requent and vigorous during the fist training sesuons and besides that the increment ofthe potentals preceded the performance of the CK and perssted during the inhibitory conditioning. The recorded potentials seem to maniest the afferent changes capable of evoking responses in areat not directly connected with the Sensory parthways andlor the integra response. Conditioning and evoked potentials Caudate nucleus Leaming mn of the information in the CN which subserved the performance of the leamed Evoked potentials IT HAS been described that both electrolytic lesions (61 and temporary blockage of the caudate nucleus (CN), by topical application of anesthetics [7], atropine or KCL3M, impair simple (Pavlovian type) and’ instrumental motor conditioned responses as well as inhibitory conditioning, while KCI injections applied to the cerebral cortex of atropine applied to the ventricle have no effects (19, 20]. ‘That the aforementioned blockage of conditioned responses is not related to apparent motor deficiencies or to ‘modification in the hunger drive has also been described {19,20}. These facts, supplemented with the results of Dean and Davis [10], of Battig er al. (4], of Chorover and Gross [9] and of other authors demonstrating the effects of lesioning the CN upon delayed and alternating condi- tioning, and the studies of Albe-Fessard et al. (1,21 regarding the polisensorial features of the CN, led us to investigate the possible relation between the evoked elec- trical activity in the CN and motor conditioning, Eight cats of either sex and with a corporal weight between 2.5 and 3.5 kg were used. The cats were trained to remain quietly on a platform placed at the end of an aisle limited by transparent plastic walls, within a soundproof chamber. In this situation a conditioning stimulus (CS), consisting of a continuous light of 1 sec duration and an intensity of approximately 38 lux, was applied. Five of the 8 cats were also submitted to generalization of the conditioning to another CS (train of 20 clicks per sec of | sec duration). If the animal walked towards the feeder (CR) situated at 20 cm from the platform at the other end of the aisle, | ml of milk was offered. Generally 26~28 associ- ations were performed daily, except on Sundays. In three animals, a series of clicks was randomly presented 500 msec after the photic. stimulation (CS) and no reward was offered, hence the animal had to suppress the motor response. This was termed inhibitory conditioning (IC). It is important to mention that the animals always had to inhibit the motor response for at least 500 msec in order to wait for the possible presentation of the clicks. Stainless steel electrodes (isolated wire of 0.25 mm dia.) were stereotaxically implanted (Kopf instrument) in the head of the CN and in the centromedian nucleus (CM) of "The authors express thei grantude to Prof. Dr Alberto Guevara Rosas and Peych, Lyzette Arditti S. for ther collaboration in the performance ofthe experiments "Present addres Laboratory of Prychophysclogcal Research, Psychology Colles, Anahuac Unwersity, Mexico (10, D.F Apdo, Postal 10-844), 100s1006 the thalamus in $ animals after the conditioned training. and before any traning in 3 cats The coordinates were A 16, L4.$ and H+ 5.5 for CN and A 5.6, L 2 Sand H+ 10 for CM after Jasper and Ajmone Marsan’s Atlas [17] ‘The electrical activity was mono- or bipolarly recorded with a c, preamplifiers (Grass P 7) with the low frequency filter set at 0.3 eps and higher frequency at 0.5 ke. The amplified signal was then recorded photographically (Grass quimographic camera) from the CRO (Tektronix 502). ‘The electrical activity was studied in the following ‘manner first, it was analyzed to determine af an evoked potential of “constant latency, with respect to the CS application, appeared. Afterwards, the percentage of evoked potential occurrences, in relation with the number ‘of daily trails, was determined. The appearance of the evoked potentials was, on occasion, measured by three Aifferent persons, independently and blindly; only & maxt- mum vanation of 4% between measurements was observed. Second, the voltage of the evoked potentials, peek to peak, was measured in photographic impressions of 10 oscilloscopic traces synchronized with the presentation of the CS (photographic means) ‘The average voltage recorded when the animals were fully conditioned was considered 100%. The mean voltage obtained in each daly session was transformed into percentage and the results were represented graphucally ‘The statistical significance of the encountered differences was analyzed by the random block test of Edwards (11] The possible correlation between the magnitude of the potentials oF their appearance with the CR was tested using the rank correlation technique (Sperman's coefficient, 21) ‘Aiter concluding the experiments, the animals were deeply anesthetized and their brains were perfused with a 10% Formalin solution injected in the left ventncle of the heart, preceded by 200 ml of a 09% saline solution. The brains wore maintamed during one week in a 10% Formalin solution after which transversal sections of 30 microns ‘width were made by means of the freezing technique The histological sections were used as negatives to make photographic impressions and in this way the location of the recording electrode tsps was established 15] GRINBERG-ZYLBERBAUM, PRADO-ALCALA AND BRUST-CARMON RESULTS ‘The behavior of the anunals was very similar t0 thai previously described [3] After a few assoctations CS-US the cats learned to sit quietly on the platform unt the CS appeared, then the eat moved towards the cup in which | ml of milk was offered. The repetition of these associations always ongimated the CR (zvamtenance) Suspending the presentation of the US caused extinction of the CR, but ‘was reestablished when the US was presented anew (re conditioning) In the preously conditioned ammals, the CS evoked in the CN and in the CM a byphasic potential, sometimes {mphasic, the first peak being negative, the second postive and the third one (if present) negative’ The average latency fof the potential was of 80 msec, although i varied in inverse relation with the intensity of the stimulus while the amplitude remained unchanged, The amphtude of these evoked potentials remained stable during the maintenance phase. The first time that the reinforcement was suppressed (extinction), the animals would leave the platform and go to the feeder when the CS appeared. However, ater a few presentations of CS without reinforcement, ‘the animals ‘would turn their heads towards the door of the chamber, or to the feeder, would then make one or two steps and stop On other occasions, they would remain on the platform. ‘The potentials evoked by the light increased considerably in this situation, but once the animal stopped responding to the CS the amplitude of the potentials decreased and then appeared rarely until finally disappearing in most of the tases, and comeiding with the disappearance of the CR. On the other hand, when the reinforcement was presented anew (reconditioning) both the behavioral responses 8s well, fa the amplitude and constancy of appearance of the potentials imereased in such a way that from the List Feconditioning sessions the behavioral and the electrical percentages were greater than 80%. Figure I shows the amplitude of the evoked potentials reconled during the ‘maintenance, extinction and reconditioning phases, The amplitude of the potential kept a close correlation with the behavioral response as can be seen in Fig 2 The same FIG. 1 The evoked potentuls i the CN ebetted by the CS, in three cats, mcreased during the mamtenance (A), decreased during the ‘extinction (B); and reappeared during the reconditioning phase (C). In this and in all suular figures each trace represen the photographie ‘im of 10 ovcloscopic trace,CONDITIONED AND EVOKED POTENTIALS IN THE CN 20 22 2 26 28 80 1007 cAUOATE NUCLEUS (CS* GREEN LIGHT —cr Bee 343088 «40a SESSIONS FIG. 2. This graph shows the varying percentage ofthe conditioned responses (slid line; the appearance of the evoked potentials inthe (CN Ginterupied line) and its voltage (dotted line) duting the maintenance, extinction and reconditioning phase, The corration was atstially significant at the level of p<0.001, figure shows how the constancy and amplitude of the evoked potentials followed the decrement and recovery of the CR. The aforementioned changes were observed in the CN and CM of the five cats except in one, where no recovery of the evoked potential was observed in the CM, during the reconditioning phase ‘The variations of the amphtude and of the rate of appearance of the evoked potentials during the different phases: maintenance, extinction and reconditioning were Statistically significant (F = 17.98 and 44.87, »<0.02 and p< 0.005), In three cats the degree of correlation between the behavior and the appearance of the potential, ranged from 0.72-0.84 with ¢ = 5.29-14.68, Between the behavior and the amplitude of the potential it ranged from 0.74~0.97 with ¢ = 6,16~34.55. This data is statistically significant at the level of p<0.001 In the animals with electrodes implanted previously to any conditioning, it was observed that as the cats acquired the CR an evoked potential similar to those already described appeared and became progressively more constant when the animal reached more than 80% correct CR. The evoked potentials remained relatively stable during the maintenance phase, while during the extinction phase they decreased considerably. The correlation analysis of the learned behavior with the appearance and amplitude of the potentials was also in all cases of 0.74~0.82 with 1 = 7,648.76, with a random occurrence probability of less than 0.001. In the S animals, originally conditioned to the light and then to an acoustical stimulus, the application of the second CS produced the appearance of the biphasic potential (negative positive) and in some cases triphasic (negative—positive—nepative), with a mean latency of 25 see. The repetition of the sound-response-reinforcement association caused an increase in this potential, which was ‘more apparent after the second conditioning session (Fig 3). The relation between the acquisition of the conditioned response on one hand, and the amplitude or appearance of the potential on the other was very close, in such a manner that the correlation analysis fluctuated between 0.94 and 0.99 (¢ = 7.61~55.0) in the different animals, with a random occurrence probability less than 0.001. In the 2 cals subjected to inhibitory conditioning it was ‘observed that the evoked potentials appeared in the CN’ independently from the animal’s movement. Furthermore the evoked potentials were of higher amplitude during the first associations in which the animals did not move towards the feeder. The histological examination showed that in all animals the tips of the electrodes implanted in the CN were placed im the head. These results, as well as those correspondent to the electrode locations in the thalamus, are illustrated in Fig4 DISCUSSION ‘The encountered correlation between the appearance and the amplitude of the evoked potential in the CN with conditioning seems to indicate that this type of electrical activity is a manifestation of some physiological process deeply related with the learning process. Simular results describing the increase of the evoked potentials have been demonstrated in other sturctures by different authors, Galambos [12, 13, 14] and Brust-Carmona et a. [5] ‘The results reported im this paper support in an important manner the participation of the CN in the008 GRINBERG-ZYLBERBAUM, PRADO-ALCALA AND BRUST-CARMONA CS: sound = Ov. 20 msec FIG. 3. The evoked potentials elicited by the sound (CS) recorded 1n the CN (3 animals), during the fist (A), second (B) and thud (C) conditionmg sessions are illustrated, Si sme 57 S8 5% FIG 4. ilustrauon of the recording sites of the evoked potenuuls eid by the conditioned stumulus. The figue was made projecting ihe histological slide upon the sereotaic alas of Jasper and Ajmone MarsanCONDITIONED AND EVOKED POTENTIALS IN THE CN integration or in the performance of motor conditioned responses as has been demonstrated by experiments in which the electrical activity of the CN was blocked inreversibly [4, 6, 9] or through pharmacological agents 119,20]. These potentials could be thought of as being related to the motor manifestations occurring during these CRs, however the appearance of the evoked potentials in the inhibitory conditioning situation and the fact that the motor conditioned response preceded the appearance or the increment of the evoked potentials in the CN deny that interpretation. Neither can the potentials be thought of as being simply the orders for the movement, since the potential is also present when the animal is inhibiting the fesponse. Therefore, it 1s more likely to thunk that this electrical change could be related with a functional incre- ment of the afferent projections to the CN. The possible visual and auditory projections to the CN have been described by Albe-Fessard et af [1,2]. These afferent 1009 projections to the CN seem to be through the polysensonal system which agrees with the fact that electrical changes related with learning processes are also observed in the ‘unspecific nuclei of the thalamus. This data supports the Postulation that the unspecific system is importantly involved in the association responsible for learning [16,18] Therefore, the CN could be one of the subcortical structures which analyses and integrates the afferent infor- ston subserving the establishment of the motor condi- tioned responses, (storage of the engram) and could also be part of the subcortical analysing structures proposed in ‘Anokhin’s mode! {3] “To conclude, these experiments give further support to the postulation that the acquisition of the instrumental motor conditioned responses depends on the activity of the different brain structures (unspecific thalamic structures, cortex, etc), but the integration appears to be in the CN. REFERENCES AlbeFesaré, D., E. 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