Interhemispheric connections of the parietal area of the cat cerebral cortex

The interhemispheric connections of the temporal cortical area in the cat cerebrum have been investigated after electrolytic coagulation of separate fields with subsequent study of the degenerated fibers course after Nauta--Gygax method. The fields 5 and 7 give origin mainly to homotopic fibers, terminating in symmetrical fields of the contralateral hemisphere. These fields also give origin to a small number of heterotopic commissural fibers, that provide bilateral connection of the fields 5 and 7 and do not get beyond the limits of the temporal cortex. The commissural fibers of the temporal cortex get into the contralateral hemisphere through the corpus callosum. In the latter, the commissural fibers of the field 5 are situated more rostral of the fibers running from the field. 7. This corresponds to topographic arrangement of the fields on the cortical surface.     

Electrocorticographic development of epileptogenic foci in the occipital region of the rat cerebral cortex

The development of cortical penicillin foci in the occipital region was studied in rats whose ages ranged from five days up to the adult age. The local application of penicillin induced the formation of an epileptogenic focus for the first time at the age of seven days. With advancing age, the amplitude of focal discharges increased, the duration of the individual components of the discharge shortened, its originally negative-positive configuration changed to a triphasic form and in the third week of life initial positivity, for a time, become the dominant component of the discharge. Projection of the discharges to the contralateral hemisphere was found to be inconstant in the second postnatal week, but appeared regularly from the age of 14 days. Synchronization of the discharges of two symmetrical foci was very poor in 7-day-old young, but improved noticeably by the 14th day; it was never complete, however, even in adulthood. The activity of symmetrical foci changed spontaneously to ECoG seizures, which were most common in 7-day-old young (in which ictal activity was usually not generalized, however) and were least frequent in 14-day-old animals. Focal discharges could not be reliably triggered by electrical stimulation of the contralateral cortex until the age of 18 days and later. The occipital part of the cortex develops somewhat later than the sensorimotor, frontal region, and during its development there also appeared phenomena which are not present in the frontal cortex.     

Effect of phenobarbital on cortical epileptogenic foci in the rat

The activity of epileptogenic foci was studied in acute experiments on adult male rats without general anaesthesia; the animals first received an intraperitoneal injection of phenobarbital [40 mg/kg] and 30 min later a cortical focus was induced by the local application of penicillin. These foci, which were induced in the sensorimotor area, had a lower discharge frequency than in the controls and the formation of projected discharges was delayed. In two symmetrical and asymmetrical cortical foci, activity was synchronized significantly more slowly after phenobarbital than in the controls. The cortical interhemispheric response was not influenced by the administered dose of phenobarbital. In these experiments, phenobarbital had an anti-epileptic effect both on the primary focus and on the spread of epileptic activity from this focus.     

Effect of diphenylhydantion on cortical epileptogenic foci in the rat

In acute experiments we studied the effect of diphenylhydantoin (DPH, 60 mg/kg i.p.) on the activity of cortical penicillin foci in 81 male rats. DPH reduced the discharge frequencies (measured by means of histograms of the intervals between adjacent discharges) and limited the projection of the discharges into non-primary cortical areas. The results for the administration of DPH before formation and after stabilization of the focus were qualitatively the same. DPH statistically significantly slowed down synchronization of the activity of two symmetrical cortical foci, but did not affect the progress of the synchronization of two asymmetrical foci, which was also very slow in the controls. DPH did not inhibit the possibility of triggering focal discharges by peripheral nerve stimulation. A detailed evaluation of focal activity allows the anti-epileptic effect of DPH to be demonstrated even in a model as potent as the penicillin focus in animals without general anaesthesia.     

Influence of clonazepam on cortical epileptogenic foci in the rat

The antiepileptic action of clonazepam was studied on epileptogenic foci induced by penicillin in sensorimotor cortex in acute experiments in rats. Clonazepam (1 mg/kg intraperitoneally) only moderately decreased the frequency of interictal discharges of single cortical focus and delayed the propagation of discharges into the ipsilateral occipital region. On the contrary, clonazepam failed to influence the callosal projection of interictal discharges in single unilateral as well as in two symmetrical foci. Spontaneous transition of interictal discharges into ictal phases regularly seen when two symmetrical foci were formed was only delayed but not blocked by clonazepam. It may be concluded that clonazepam exhibits only a weak anticonvulsant action against cortical foci and against secondary generalization of epileptic activity.     

Intra- and Interhemispheric Propagation of Electrophysiological Synchronous Activity and Its Modulation by Serotonin in the Cingulate Cortex of Juvenile Mice

Disinhibition of the cortex (e.g., by GABA -receptor blockade) generates synchronous and oscillatory electrophysiological activity that propagates along the cortex. We have studied, in brain slices of the cingulate cortex of mice (postnatal age 14–20 days), the propagation along layer 2/3 as well as the interhemispheric propagation through the corpus callosum of synchronous discharges recorded extracellularly and evoked in the presence of 10 μM bicuculline by electrical stimulation of layer 1. The latency of the responses obtained at the same distance from the stimulus electrode was longer in anterior cingulate cortex (ACC: 39.53 ± 2.83 ms, n = 7) than in retrosplenial cortex slices (RSC: 21.99 ± 2.75 ms, n = 5; p<0.05), which is equivalent to a lower propagation velocity in the dorso-ventral direction in ACC than in RSC slices (43.0 mm/s vs 72.9 mm/s). We studied the modulation of this propagation by serotonin. Serotonin significantly increased the latency of the intracortical synchronous discharges (18.9% in the ipsilateral hemisphere and 40.2% in the contralateral hemisphere), and also increased the interhemispheric propagation time by 86.4%. These actions of serotonin were mimicked by the activation of either 5-HT_1B or 5-HT_2A receptors, but not by the activation of the 5-HT_1A subtype. These findings provide further knowledge about the propagation of synchronic electrical activity in the cerebral cortex, including its modulation by serotonin, and suggest the presence of deep differences between the ACC and RSC in the structure of the local cortical microcircuits underlying the propagation of synchronous discharges.     

Auditory and Visual Interhemispheric Communication in Musicians and Non-Musicians

The corpus callosum (CC) is a brain structure composed of axon fibres linking the right and left hemispheres. Musical training is associated with larger midsagittal cross-sectional area of the CC, suggesting that interhemispheric communication may be faster in musicians. Here we compared interhemispheric transmission times (ITTs) for musicians and non-musicians. ITT was measured by comparing simple reaction times to stimuli presented to the same hemisphere that controlled a button-press response (uncrossed reaction time), or to the contralateral hemisphere (crossed reaction time). Both visual and auditory stimuli were tested. We predicted that the crossed-uncrossed difference (CUD) for musicians would be smaller than for non-musicians as a result of faster interhemispheric transfer times. We did not expect a difference in CUDs between the visual and auditory modalities for either musicians or non-musicians, as previous work indicates that interhemispheric transfer may happen through the genu of the CC, which contains motor fibres rather than sensory fibres. There were no significant differences in CUDs between musicians and non-musicians. However, auditory CUDs were significantly smaller than visual CUDs. Although this auditory-visual difference was larger in musicians than non-musicians, the interaction between modality and musical training was not significant. Therefore, although musical training does not significantly affect ITT, the crossing of auditory information between hemispheres appears to be faster than visual information, perhaps because subcortical pathways play a greater role for auditory interhemispheric transfer.     

Mechanisms of transmission of excitation grnerated in the orbito-frontal cortex]

Experiments carried out on cats showed that interhemispheric generalization of strychnine spasmodic potentials from the orbito-frontal cortex were mainly brought about by the callosal pathway. In contrast to other projection and association zones of the cortex only the frontal portion of the corpus callosum (the rostrum) and not the whole corpus took part in transmitting strychnine commissures. The results of investigations carried out were consistent with the general conception of the principle of the determining dispatch station (DDS) in the nervous system integration activity. These results reveal that the appearance of the DDS induces secondary excitation foci which fully reproduce tha work regimen of the DDS. Switching of such "mirror" foci representing "the destination stations" from the DDS influence led to the elimination of the activity induced in them.     

Relevance of the callosal transfer in defining the peripheral reactivity of somesthetic cortical neurones

Experiments were performed in 13 chloralose-anaesthetized, curarized cat preparations (monitoring of rectal temperature, heart rate, expired pCO2 and EEG), in order to ascertain whether, and to what extent, the reactivity to ipsilateral skin shocks of the neurones of the anterior ectosylvian and anterior suprasylvian gyri (AEG and ASG, respectively) is dependent on the callosal output of the somatosensory areas of the contralateral hemisphere. Indeed, we knew from previous experiments that a high proportion of AEG and ASG neurones having bilateral peripheral receptive fields (PRFs) can be excited by direct stimulation of the contralateral homonymous areas, and that the callosal fibres originating in the latter carry somesthetic impulses related to contralateral PRFs. A preliminary analysis was carried out on the amplitude and latency relationships between the evoked potentials (EP) recorded simultaneously from the two hemispheres and from the corpus callosum (CC) following stimulation of the forepaw of one side. The results obtained showed good correlations between the onset and development of the EPs picked up from the hemisphere ipsilateral to the stimulated skin, on the one hand, and onset and development of the EPs recorded from the contralateral hemisphere and the corpus callosum, on the other. At a further stage of the experiments, the EPs elicited upon ipsilateral and contralateral skin shocks in the AEG-ASG area have been recorded and averaged before, during and after the reversible inactivation of callosal somesthetic transmission. This was achieved by applying polarizing currents (0.2-1 mA) to the rostral portion of the CC, adequacy and reversibility of this method having been tested by observing, respectively, suppression and prompt restoration of transcallosal EPs and of the asynaptic spiking produced by cortical cells when antidromically invaded from contralateral homotopic cortex. It was seen that during CC blockade the EPs elicited in the AEG-ASG areas did not show any change either in amplitude or time-course if brought about by contralateral peripheral stimulation, whereas those evoked by ipsilateral skin shocks exhibited significant reduction, which was related to the strength of CC polarization and to the reduction of transcallosal EPs. In control experiments similar effects were observed after ablation of somatosensory areas of the hemisphere which send off somesthetic callosal impulses, whereas strychninization of these areas caused effects opposite in sign, i.e., enhancement of the ipsilateral but not of the contralateral EPs in the areas of the untreated hemisphere. By testing the effects of CC polarization on single AEG-ASG neurones, it was observed that the responses of units linked only with contralateral PRFs (Group I; 7 units tested) were unaffected by callosal polarization. The discharges of neurones provided with wide and bilateral PRFs (Group II; 27 units tested) were not affected if elicited by contralateral PRF shocks but were deeply impaired (in 11 neurones out the 27) when provoked by ipsilateral PRF stimulation. The effect consisted chiefly of the disappearance of the first high peak of the PSTHs, and when recording intracellularly graded events, it was mirrored by a large decrease of the postsynaptic excitatory potentials elicited in Group II neurones by ipsilateral PRF shocks. A late scattered histographic component was identified in the PSTHs of such cells, which did not appear to be significantly altered during CC blockade. These effects were observed on the ipsilateral responses of 11 out of the 27 Group II neurones so tested whereas the ipsilateral PSTHs of the remaining 16 Group II neurones either did not undergo significant changes during the callosal blockade or escaped evaluation because of high spontaneous shifts of neural responsiveness. The results are discussed mainly with a view to the possible functional role of the specific somesthetic callosal fibres in defining ipsilateral reactivity for the wide-field cells of the AEG-ASG area.     

The interhemispheric asymmetry of the spatial-temporal organization of cortical potentials in the rabbit]

In experiments on freely moving rabbits the existence of interhemispheric asymmetry of spatial-temporal organization of the cortical potentials in the state of calm alertness was shown. Episodes were revealed of interhemispheric discordance of spatial reconstructions of momentary reliefs of the cortical potentials topograms 1/64-1/3-1/5 s in duration, i.e. periods of the theta- or delta-rhythms. Total duration of time of interhemispheric asymmetry (according to significant divergences in dynamics of resemblance coefficients of consequent reliefs of the cortical potentials topograms) in most cases was not less than 10% and not more than 30% of time of analysis epoch and consisted of discordance episodes both of different global reconstructions of spatial correlations of the left and right hemisphere potentials and of interhemispheric local shifts in spatial organization of the cortex potentials.     

A function of the corpus callosum in the Siamese cat

Recordings were made from 180 single cells, or cell clusters, in the Clare-Bishop area of two Siamese cats. In one cat, the corpus callosum had been sectioned prior to recording and all cells were driven by the contralateral eye exclusively. In the other recordings were made before and after callosal section. Before callosal section, most cells were binocularly driven but dominated by the contralateral eye. There were striking examples of binocular interaction and some cells could only be activated by simultaneous binocular stimulation. After callosal section, cells were driven by the contralateral eye only. The same experiment performed in a normal cat revealed no change in binocularity following section of the corpus callosum. We conclude that one of the functions of the corpus callosum in the Siamese cat is to generate binocular neurons.     

Wnt/calcium signaling mediates axon growth and guidance in the developing corpus callosum

It has been shown in vivo that Wnt5a gradients surround the corpus callosum and guide callosal axons after the midline (postcrossing) by Wnt5a-induced repulsion via Ryk receptors. In dissociated cortical cultures we showed that Wnt5a simultaneously promotes axon outgrowth and repulsion by calcium signaling. Here to test the role of Wnt5a/calcium signaling in a complex in vivo environment we used sensorimotor cortical slices containing the developing corpus callosum. Plasmids encoding the cytoplasmic marker DsRed and the genetically encoded calcium indicator GCaMP2 were electroporated into one cortical hemisphere. Postcrossing callosal axons grew 50% faster than pre-crossing axons and higher frequencies of calcium transients in axons and growth cones correlated well with outgrowth. Application of pharmacological inhibitors to the slices showed that signaling pathways involving calcium release through IP3 receptors and calcium entry through TRP channels regulate post-crossing axon outgrowth and guidance. Co-electroporation of Ryk siRNA and DsRed revealed that knock down of the Ryk receptor reduced outgrowth rates of postcrossing but not precrossing axons by 50% and caused axon misrouting. Guidance errors in axons with Ryk knockdown resulted from reduced calcium activity. In the corpus callosum CaMKII inhibition reduced the outgrowth rate of postcrossing (but not precrossing) axons and caused severe guidance errors which resulted from reduced CaMKII-dependent repulsion downstream of Wnt/calcium. We show for the first time that Wnt/Ryk calcium signaling mechanisms regulating axon outgrowth and repulsion in cortical cultures are also essential for the proper growth and guidance of postcrossing callosal axons which involve axon repulsion through CaMKII.     

Anatomy of corpus callosum in prenatally malnourished rats

The effect of prenatal malnutrition on the anatomy of the corpus callosum was assessed in adult rats (45-52 days old). In the prenatally malnourished animals we observed a significant reduction of the corpus callosum total area, partial areas, and perimeter, as compared with normal animals. In addition, the splenium of corpus callosum (posterior fifth) showed a significant decrease of fiber diameters in the myelinated fibers without changing density. There was also a significant decrease in diameter and a significant increase in density of unmyelinated fibers. Measurements of perimeter's fractal dimensions from sagittal sections of the brain and corpus callosum did not show significant differences between malnourished and control animals. These findings indicate that cortico-cortical connections are vulnerable to the prenatal malnutrition, and suggest this may affect interhemispheric conduction velocity, particularly in visual connections (splenium).     

Bilateral receptive fields in cortical area SII: contribution of the corpus callosum and other interhemispheric commissures

The corpus callosum contributes to the interhemispheric transfer of somatosensory information. Since the somatosensory pathways are essentially crossed, a number of studies have postulated that the corpus callosum may be responsible for the presence of bilateral receptive fields (RFs) in cortical area SII. Moreover, subcortical structures, as well as some of the other commissures, may also contribute to the bilateral nature of these cells. In order to assess the relative importance of the corpus callosum, this study compared the RF properties of cells in area SII of callosum-sectioned cats to normal cats, using single-cell recordings. Results showed that the corpus callosum makes an important contribution to the bilateral activation of cells in SII, since the proportion of cells with bilateral RFs found in callosum-sectioned cats was less than half that obtained in normal cats. The decrease in the proportion of bilateral RFs was found for all body regions with the exception of the face. However, the substantial number of bilateral RFs remaining in callosotomized cats indicates that this structure is not the sole contributor to the bilateral activation of cells in SII. In order to determine whether this residual bilateral activation might be mediated by the other interhemispheric commissures, a group of cats was subjected, besides the callosotomy, to the additional transection of their subcortical commissures, including the anterior, posterior, habenular, and intertectal commissures, as well as the massa intermedia. When this group of deep-split cats was compared to the callosotomized group, the results indicated that the contribution of the other commissures to bilateral activation is negligible, since approximately the same proportion of bilateral RFs was encountered in the two groups. The relative importance of the callosal contribution to bilateral RFs of different body regions is discussed with respect to the roles commonly attributed to this structure.     

<Emphasis Type="Italic">PLP1</Emphasis> Gene Variation Modulates Leftward and Rightward Functional Hemispheric Asymmetries

Molecular neurobiological factors determining corpus callosum physiology and anatomy have been suggested to be one of the major factors determining functional hemispheric asymmetries. Recently, it was shown that allelic variations in two myelin-related genes, the proteolipid protein 1 gene PLP1 and the contactin 1 gene CNTN1, are associated with differences in interhemispheric integration. Here, we investigated whether three single nucleotide polymorphisms that were associated with interhemispheric integration via the corpus callosum in a previous study also are relevant for functional hemispheric asymmetries. To this end, we tested more than 900 healthy adults with the forced attention dichotic listening task, a paradigm to assess language lateralization and its modulation by cognitive control processes. Moreover, we used the line bisection task, a paradigm to assess functional hemispheric asymmetries in spatial attention. We found that a polymorphism in PLP1, but not CNTN1, was associated with performance differences in both tasks. Both functional hemispheric asymmetries and their modulation by cognitive control processes were affected. These findings suggest that both left and right hemisphere dominant cognitive functions can be modulated by allelic variation in genes affecting corpus callosum structure. Moreover, higher order cognitive processes may be relevant parameters when investigating the molecular basis of hemispheric asymmetries.     

The cortical EEG in the acallosal mouse

The congenital absence of the corpus callosum has recently been found to occur among some mice of the ddN strain in our laboratory. Morphologically, the absence of corpus callosum was classified into two types. One was complete agenesis of corpus callosum, and the other revealed partial agenesis (genu or splenium, or both). In this experiment, the differences of cortical EEG spectral characteristics were studied on the normal, hypogenesis and acallosal mice. A total of 75 male and female adult mice were used. Under light ether anesthesia, five stainless steel electrodes (0.29 mm) were implanted stereotaxically in the bilateral cerebral cortex and cerebellum at a depth of 1.0 mm. After one week recovery period, the correlations of EEG of five combinations, such as homotopic anterior vs. posterior, left vs. right hemispheres, crossed contralateral, and homotopic anterior and posterior to the reference electrode, were analysed with a signal processor. In the complete acallosal mice (n = 11), the correlation coefficients were significantly lower (p less than 0.05), in the crossed contralateral, left vs. right hemispheres, and homotopic posterior to reference. In the hypogenesis of corpus callosum (n = 7), however, they were no significant differences in the correlation compared with normal mice (n = 57). At the end of experiment, the brain was removed and fixed with 10% formalin, then it was cut in half along the midsagittal plane. The midsagittal section was used to examine an outline of the corpus callosum.     

Propagation of epileptiform events across the corpus callosum in a cingulate cortical slice preparation

We report on a novel mouse in vitro brain slice preparation that contains intact callosal axons connecting anterior cingulate cortices (ACC). Callosal connections are demonstrated by the ability to regularly record epileptiform events between hemispheres (bilateral events). That the correlation of these events depends on the callosum is demonstrated by the bisection of the callosum in vitro. Epileptiform events are evoked with four different methods: (1) bath application of bicuculline (a GABA-A antagonist); (2) bicuculline+MK801 (an NMDA receptor antagonist), (3) a zero magnesium extracellular solution (0Mg); (4) focal application of bicuculline to a single cortical hemisphere. Significant increases in the number of epileptiform events, as well as increases in the ratio of bilateral events to unilateral events, are observed during bath applications of bicuculline, but not during applications of bicuculline+MK-801. Long ictal-like events (defined as events >20 seconds) are only observed in 0Mg. Whole cell patch clamp recordings of single neurons reveal strong feedforward inhibition during focal epileptiform events in the contralateral hemisphere. Within the ACC, we find differences between the rostral areas of ACC vs. caudal ACC in terms of connectivity between hemispheres, with the caudal regions demonstrating shorter interhemispheric latencies. The morphologies of many patch clamped neurons show callosally-spanning axons, again demonstrating intact callosal circuits in this in vitro preparation.     

Hemispheric interaction in man during perception of visual stimuli]

Averaged evoked potentials (AEP) to verbal (letters) and nonverbal (random shapes) stimuli exposed in the left and right visual fields were registered in healthy subjects with normal vision. Analysis of the later AEP latencies pointed to asymmetry in the temporal parameters of the interhemispheric interaction. The late AEP latency is shorter in the right hemisphere than in the left hemisphere. The difference is more pronounced in responses to nonverbal stimuli. The earlier development of the evoked potential in the right hemisphere (or the later one in the left hemisphere) accounts for the interhemispheric difference in the temporal parameters of the late AEP components. Comparison of the latency of the component P300 to verbal and nonverbal stimuli presented in the ipsilateral or the contralateral visual fields reveals a transfer of the results of the cortical processing of visual information in the course of interhemispheric interaction.     

Glutamate transport,glutamine synthetase and phosphate-activated glutaminase in rat CNS white matter. A quantitative study

Glutamatergic signal transduction occurs in CNS white matter, but quantitative data on glutamate uptake and metabolism are lacking. We report that the level of the astrocytic glutamate transporter GLT in rat fimbria and corpus callosum was approximately 35% of that in parietal cortex; uptake of [3H]glutamate was 24 and 43%, respectively, of the cortical value. In fimbria and corpus callosum levels of synaptic proteins, synapsin I and synaptophysin were 15-20% of those in cortex; the activities of glutamine synthetase and phosphate-activated glutaminase, enzymes involved in metabolism of transmitter glutamate, were 11-25% of cortical values, and activities of aspartate and alanine aminotransferases were 50-70% of cortical values. The glutamate level in fimbria and corpus callosum was 5-6 nmol/mg tissue, half the cortical value. These data suggest a certain capacity for glutamatergic neurotransmission. In optic and trigeminal nerves, [3H]glutamate uptake was < 10% of the cortical uptake. Formation of [14C]glutamate from [U-14C]glucose in fimbria and corpus callosum of awake rats was 30% of cortical values, in optic nerve it was 13%, illustrating extensive glutamate metabolism in white matter in vivo. Glutamate transporters in brain white matter may be important both physiologically and during energy failure when reversal of glutamate uptake may contribute to excitotoxicity.