[Retinal and Cortical Activation by Electrical Stimulation with Retina Implants]. / Retinale und kortikale Aktivierung durch elektrische Stimulation mit Netzhautimplantaten.

In Germany, about 30,000 to 40,000 people suffer from retinitis pigmentosa (RP), which ultimately results in blindness. The only aid to blind RP patients are retinal implants: These have been under development for several years and have now been approved as a medical product. Retinal implants produce visual perceptions in response to electrical stimulation of the degenerated retina and are useful in the everyday life of blind people. However, the currently achievable quality of vision is such that people with a retinal implant are still legally blind. The visual quality that can be achieved with epi- and subretinal implants depends not only on patient-specific factors such as individual history and status of retinal degeneration, but especially on the interface between implant and retina and the quality of the achievable neuronal activation. Biophysical approaches to functional improvements of the implants are founded on the physiology of the retina (cell density, intraretinal interconnections), are based on technical optimisation of the interface (electrode materials, size and density), and exploit the stimulation protocols with which visual information is fed into the degenerated retina (time courses of electrical stimuli, spatiotemporal stimulation pattern). Optimisation of stimulation parameters can be supported by a detailed analysis of cortical responses, with appropriate electrophysiological and optical methods. This article looks at both the physiological and biophysical fundamentals of electrical retinal stimulation, as well as the resulting retinal and cortical activation.

Metaplasticity of horizontal connections in the vicinity of focal laser lesions in rat visual cortex.

Focal cortical injuries are accompanied by a reorganization of the adjacent neuronal networks. An increased synaptic plasticity has been suggested to mediate, at least in part, this functional reorganization. Previous studies showed an increased long-term potentiation (LTP) at synapses formed by ascending fibres projecting onto layers 2/3 pyramidal cells following lesions in rat visual cortex. This could be important to establish new functional connections within a vertical cortical column. Importantly, horizontal intracortical connections constitute an optimal substrate to mediate the functional reorganization across different cortical columns. However, so far little is known about their potential implication in the functional rewiring post-lesion. Here, we investigated possible alterations of synaptic plasticity of horizontal connections in layers 2/3 in an 'ex vivo-in vitro' model of focal laser lesion in rat visual cortex. LTP at these synapses was found to be enhanced post-lesion, whereas long-term depression (LTD) was impaired, revealing a metaplastic shift toward strengthening of these synapses. Furthermore, we disclosed a prolonged decay-time constant of NMDAR-dependent currents, which can contribute to the enhanced LTP. Taken together these data revealed that a laser lesion-induced focal damage of the visual cortex is accompanied by a facilitated potentiation of horizontal synaptic connections in the vicinity of the focal injury. This specific strengthening of synaptic plasticity at horizontal connections in layers 2/3 might be one important cellular mechanism to compensate focal injury-mediated dysfunction in the cerebral cortex.

Functional selectivity of interhemispheric connections in cat visual cortex.

The functional specificity of callosal connections was investigated in visual areas 17 and 18 of adult cats, by combining in vivo optical imaging of intrinsic signals with labeling of callosal axons. Local injections of neuronal tracers were performed in one hemisphere and eight single callosal axons were reconstructed in the opposite hemisphere. The distributions of injection sites and callosal axon terminals were analyzed with respect to functional maps in both hemispheres. Typically, each callosal axon displayed 2 or 3 clusters of synaptic boutons in layer II/III and the upper part of layer IV. These clusters were preferentially distributed in regions representing the same orientation and the same visuotopic location as that at the corresponding injection sites in the opposite hemisphere. The spatial distribution of these clusters was elongated and its main axis correlated well with the preferred orientation at the injection site. These results demonstrate a specific organization of interhemispheric axons that link cortical regions representing the same orientation and the same location of visual stimuli. Visual callosal connections are thus likely involved in the processing of coherent information in terms of shape and position along the midline of the visual field, which may facilitate the fusion of both hemifields into the percept of a single visual scene.

Neuroscience. Noise makes sense in neuronal computing.

The united efforts of assemblies of neurons in the brain's primary visual cortex translate incoming visual signals into action potentials. These action potentials encode, for example, the contrast and orientation of different parts of the image. Some neurons are sensitive to one particular orientation, other are sensitive to other orientations, but all neurons respond equally well to the image contrast. In a Perspective, Volgushev and Eysel explain the finding (Anderson et al.) that neurons are able to maintain this sensitivity to the orientation of a stimulus regardless of the contrast by adding noise to the membrane potential, such that action potentials can also be generated in response to weak signals at low contrast.

Context, state and the receptive fields of striatal cortex cells.

Visual cortical cells are commonly characterized by their receptive-field structure. Originally, a visual receptive field was defined in a purely spatial way as that retinal area from which a change in spiking response of the regarded cell could be elicited by visual stimulation. The first attempts to understand receptive-field structure were based entirely on the anatomical connectivity of the primary visual pathway. More recently, however, it has been discovered that the spatial and temporal context in which a stimulus is presented to a cell can strongly influence its receptive field, and this in turn is dependent on the state of arousal and attention. Accordingly, new concepts recognize that cortical receptive fields are highly dynamic entities embracing more than the sum of the full spatial and temporal response properties of a cell.

Topographical Aspects of Intracortical Excitation and Inhibition Contributing to Orientation Specificity in Area 17 of the Cat Visual Cortex.

Intracortical mechanisms contributing to orientation and direction specificity were investigated with a method of local cortical inactivation. Single-unit activity was recorded in area 17 of the anaesthetized cat while a small volume of cortical tissue 400 - 2900 microm lateral to the recorded cell was inactivated by gamma-aminobutyric acid (GABA) microiontophoresis. Cells were stimulated with moving bars of variable orientation and changes of the response were monitored. Recording and inactivation sites were histologically verified. Statistically significant changes in orientation tuning during GABA-induced remote inactivation were observed in 80 of 145 cells (55%), and consisted in a reduced orientation specificity due to either increased (36%) or decreased (19%) responses. Increases of responses were more pronounced for the non-optimal orientations. This effect mainly occurred with GABA application at distances around 500 microm and is interpreted as loss of inhibition. Reduced orientation specificity as a result of decreasing response mainly to the optimal orientation was interpreted as loss of excitation. This effect most frequently occurred with inactivation at distances around 1000 microm. Loss of inhibition was also elicited from a distance of 1000 microm; such inhibition, however, affected only directionality, without inducing changes in orientation tuning. For several cells at distances >1000 microm from the inactivation site a temporal sequence consisting of a change in direction specificity followed by a reduction of orientation specificity, and finally by direct GABAergic inhibition of the cell under study, could be induced with gradually increasing ejecting currents. The results indicate that excitation and inhibition originating from populations of neurons at different horizontal distances differentially contribute to direction and orientation specificity of a given visual cortical cell.

The effects of partial retinal lesions on activity and size of cells in the dorsal lateral geniculate nucleus.

The nasal parts of the adult cat retina were photocoagulated. In the dorsal lateral geniculate nucleus (dLGN) the projection of the remaining innervation was shown by anterograde transport of 3H-proline after eye injections. The neuronal activity was measured from single cells across the border region between innervated and deafferented parts of layer A of the dLGN contralateral to the lesions. A gradual decrease from normal light-excitability to total inexcitability was observed over a range of 300 micron. The perikaryal cell sizes measured in the same part of the dLGN displayed a concomitant decrease. Blockage of the afferent impulses by chronic application of tetrodotoxin did not change the results, suggesting that it is the loss of connections, not the loss of activity, that produces the transneuronal atrophy in the adult cat dLGN.

Dendritic plasticity in the early postnatal feline retina: quantitative characteristics and sensitive period.

Retinal lesions were made in kittens between 3 and 60 days postnatal age and in adult cats. After postlesion survival times ranging from 4 to 11 months the dendritic morphology of retinal ganglion cells was revealed by retrograde labeling with horseradish peroxidase or with neurofibrillar staining techniques. After retinal lesions on the third postnatal day changes of dendritic morphology were observed in retinal ganglion cells adjacent to regions of retrograde degeneration. Originating from eccentrically positioned somata the dendritic fields extended into the regions that were free of neighboring cells. The dendrites oriented toward the ganglion-cell-free region were elongated and thicker than normal. The density of dendrites per unit area was increased in this part of the dendritic trees. Lesions on the 20th, 38th, and 56th postnatal days elicited increasingly weaker changes of dendritic morphology. The sensitive period for the type of dendritic plasticity described ends between 40 and 60 days postnatally.

Network of GABAergic large basket cells in cat visual cortex (area 18): implication for lateral disinhibition.

Anatomical and immunohistochemical data indicate that, in addition to pyramidal neurons, nonpyramidal cells are exposed to perisomatic inhibition mediated by gamma-aminobutyric acid (GABA)-containing terminals. However, no direct information is available as yet for the origin of GABAergic inputs to morphologically identified GABAergic neurons. In the present paper, we studied the topographical and synaptic relationship between identified GABAergic large basket cells and their immunohistochemically characterized target neurons revealed by parvalbumin-(PV) and GABA immunostaining in the same material. Extracellularly applied biocytin labelled a total of 36 and 9 large basket cells in layers III and V, respectively. Of these, the axonal arborizations of two basket cells, BC1 and BC2, were reconstructed. The axon of BC1 occupied an area of about 2.3 x 2.2 mm2 in layer III, providing a total of 2,755 terminals. The axon of BC2 showed an overall extent of 3.8 x 1.7 mm2 in layer V elongated in the anteroposterior direction, and gave off 1,599 terminals. Immunostaining for PV was carried out to reveal putative nonpyramidal targets for BC1 and BC2. It was found that in addition to immunonegative cells, they established an average of 4-6 perisomatic contacts onto each of 58 (BC1) and 33 (BC2) PV-immunopositive neurons. For electron microscopic verification, 23 terminals apposing the somata of 12 PV-immunopositive neurons were selected. Each terminal was found to establish symmetrical (type II) contacts with its targeted cell. Furthermore, the distribution of soma area of the targeted PV-immunopositive cells and of identified large basket cells showed remarkable similarity, implying that the two populations were actually the same. In addition, the average horizontal distance between neighbouring PV-immunopositive target cells was found to be about 100 microns both in layers III and V. The results suggest that in area 18 the same large basket cell provides direct inhibition to certain pyramidal cells and facilitation to other pyramidal neurons, by inhibiting their presynaptic large basket cells at regular intervals.

Immunohistochemical studies on neurofilamentous hypertrophy in degenerating retinal terminals of the olivary pretectal nucleus in the rat.

Following section of the optic nerve, degenerating retinal terminals reveal an accumulation of neurofilaments (neurofilamentous hypertrophy) as demonstrated by silver impregnation techniques or electron microscopy. The present study examined degenerating retinal terminals by means of immunohistochemistry and antibodies specific for the triplet of neurofilament proteins of low (NF-L), medium (NF-M), and high (NF-H) molecular weight class. Following unilateral optic nerve section in the rat and survival of 1, 2, 4, 8, and 21 days, brains were perfused with aldehyde fixative, sliced on a vibratome and stained for neurofilaments by using the peroxidase-antiperoxidase technique. Other brains were frozen, cut in the native state, and slide-mounted sections were fixed by acetone. Side comparisons in visual pathways were made in frontal sections, taking advantage of the near complete crossing of retinal fibers in the rat. Anterograde degeneration of axons occurred in the optic tract and branchium colliculi. Changes of terminals were investigated in the olivary pretectal nucleus, which contains a dense aggregation of retinal terminals in the core region. The optic tract and branchium colliculi showed a reduction in immunostaining for neurofilament proteins following axotomy. Within the core region of the olivary pretectal nucleus, strong increases of immunoreactivity of NF-L and NF-M were detected beginning at 2 days postlesion and persisting at 8 days. No changes in NF-H proteins were found in the terminal regions with three different antibody probes. The increase in immunostaining reflects the accumulation of neurofilament proteins in the degenerating retinal terminals, i.e., neurofilamentous hypertrophy.(ABSTRACT TRUNCATED AT 250 WORDS)

Axonal topography of cortical basket cells in relation to orientation, direction, and ocular dominance maps.

The axonal (bouton) distributions of a layer 4 clutch cell (CC), two layer 3 medium-sized basket cells (MBC), and a layer 3 large basket cell (LBC) to orientation, direction, and ocular dominance maps were studied quantitatively. 1) The CC provided exclusively local projections (<380 microm from the soma) and contacted a narrow "niche" of functional representations. 2) The two MBCs emitted local projections (75% and 79% of all boutons), which were engaged with isoorientations (61% and 48%) and isodirections, and long-range projections (25% and 21%, >313 microm and >418 microm), which encountered cross-orientation sites (14% and 12%) and isoorientation sites (7% and 5%). Their direction preferences were mainly perpendicular to or opposite those of local projections. 3) The LBC provided the majority (60%) of its boutons to long-range distances (>437 microm). Locally, LBC boutons showed a rather balanced contribution to isoorientations (19%) and cross-orientations (12%) and preferred isodirections. Remotely, however, cross-orientation sites were preferred (31% vs. 23%) and the directional output was balanced. 4) Monte Carlo simulations revealed that the differences between the orientation specificity of local and long-range projections cannot be explained by a homogeneous lateral distribution of the boutons. 5) There was a similar eye preference in the local and long-range projection fields of the MBCs. The LBC contacted both contra- and ipsilateral eye domains. 6) The basket axons showed little laminar difference in orientation and direction topography. The results suggest that an individual basket cell can mediate a wide range of effects depending on the size and termination pattern of the axonal field.

Investigation of cortical reorganization in area 17 and nine extrastriate visual areas through the detection of changes in immediate early gene expression as induced by retinal lesions.

The effect of binocular central retinal lesions on the expression of the immediate early genes c-fos and zif268 in the dorsal lateral geniculate nucleus (dLGN) and the visual cortex of adult cats was investigated by in situ hybridization and immunocytochemistry. In the deafferented region of the dLGN, the c-fos mRNA level was decreased within 3 days. The dimensions of the geniculate region showing decreased amounts of c-fos mRNA matched the predictions based on the lesion size and the retinotopic maps of Sanderson ([1971] J. Comp. Neurol. 143:101-118). We did not detect zif268 mRNA in the dLGN. At the cortical level, both c-fos and zif268 mRNA expression decreased in the sensory-deprived region of area 17. In addition, the portions of areas 18, 19, 21a, 21b, and 7, as well as the posterior medial lateral suprasylvian area, the posterior lateral lateral suprasylvian area, the ventral lateral suprasylvian area, and the dorsal lateral suprasylvian area corresponding to the retinal lesions also displayed decreased c-fos and zif268 mRNA levels. Immunocytochemistry revealed similar changes for Zif268 and Fos protein. Three days post lesion, the dimensions of the lesion-affected cortical loci exceeded the predictions in relation to the size of the retinal lesions and the available retinotopic maps. Longer postlesion survival times clearly resulted in a time-dependent restoration of immediate early gene expression from the border to the center of the lesion-affected cortical portions. Our findings represent a new approach for investigating the capacity of adult sensory systems to undergo plastic changes following sensory deprivation and for defining the topographic nature of sensory subcortical and cortical structures.

Effect of sensory deafferentation on immunoreactivity of GABAergic cells and on GABA receptors in the adult cat visual cortex.

To investigate the effects of sensory deafferentation on the cortical GABAergic circuitry in adult cats, glutamic acid decarboxylase (GAD) and gamma-aminobutyric acid (GABA) immunoreactivity and GABA receptor binding were studied in the visual cortex of normal cats and compared with cats that had received restricted binocular central lesions of the retina and had survived for 2 weeks postlesion in a normal visual environment. In the visual cortex of lesioned cats, two changes were observed in the number of GAD-immunoreactive elements in the regions affected by the retinal lesions: the number of GAD-positive puncta decreased, whereas that of GAD-immunoreactive somata increased. In contrast, no detectable changes were measured in the number of GABA-immunopositive somata or puncta. At the receptor level, we observed no differences in either the laminar distribution or the affinity of cortical GABAA and GABAB receptors labeled with [3H]-muscimol and [3H]-baclofen, respectively, in the lesioned versus normal cats. We present the hypothesis that sensory deafferentation in these adult cats (1) leads to a reduction of cortical GABAergic inhibition in the deafferented region, and (2) that this decreased inhibition may permit changes in efficiency of synapses and (3) that these changes may represent a first stage of events underlying the retinotopic reorganization preceeding the structural changes.

Cellular organization of reciprocal patchy networks in layer III of cat visual cortex (area 17).

There is no direct information available concerning the exact spatial characteristics of long-range axons and their relationship with the patchy phenomena observed after extracellular injection of retrograde tracers. In the present study, using the recently introduced neuronal tracer biocytin, we demonstrate by detailed three-dimensional reconstruction of 10 pyramidal cells in layer III, that their clustered axonal terminals form a specific patchy network in layers II and III. The reconstructed network occupied an area of 6.5 x 3.5 mm parallel to the cortical surface elongated in an anteroposterior direction. The average centre-to-centre distance between patches within the network was 1.1 mm. On average, the axonal field of each of the 10 pyramidal cells contained a total of 417 boutons at four to eight distinct sites (patches), and in each patch, an average of 79 boutons was provided by the same cell. The identified connections between the patches were predominantly reciprocal. Detailed analyses have shown that many pyramidal cells of the network are directly interconnected so that each of them can receive one to four, chiefly axospinous, contacts onto the distal segment of its apical and basal dendrites from the axon of another pyramidal cell belonging to a different patch labelled from the same injection site. We hypothesize that the possible functional role of the network is to link remote sites with similar physiological characteristics, such as orientation preference, supporting the model of Mitchison and Crick [(1982) Proc. natn. Acad. Sci. U.S.A. 79, 3661-3665].

Spatio-temporal plasticity of cortical receptive fields in response to repetitive visual stimulation in the adult cat.

Many psychophysical experiments on perceptual learning in humans show increases of performance that are most probably based on functions of early visual cortical areas. Long-term plasticity of the primary visual cortex has so far been shown in vivo with the use of visual stimuli paired with electrical or pharmacological stimulation at the cellular level. Here, we report that plasticity in the adult visual cortex can be achieved by repetitive visual stimulation. First, spatial receptive field profiles of single units (n=38) in area 17 or 18 of the anesthetized cat were determined with optimally oriented flashing light bars. Then a conditioning protocol was applied to induce associative synaptic plasticity. The receptive field center and an unresponsive region just outside the excitatory receptive field were synchronously stimulated ('costimulation', repetition rate 1 Hz; for 10-75 min). After costimulation the receptive field and its adjacent regions were mapped again. We observed specific increases of the receptive field size, changes of the receptive field subfield structure as well as shifts in response latency. In 37% of the cells the receptive field size increased specifically towards the stimulated side but not towards the non-stimulated opposite side of the receptive field. In addition, changes in the relative strength and size of the on and off subfield regions were observed. These specific alterations were dependent on the level of neuronal activity during costimulation. During recovery, the new responses dropped down to 120% of the preconditioning value on average within 103 min; however, the decay times significantly depended on the response magnitude after costimulation. In the temporal domain, the latency of new responses appeared to be strongly influenced by the latency of the response during costimulation.Twenty-nine percent of the units displayed no receptive field enlargement, most likely because the activity during costimulation was significantly lower than in the cases with enlarged receptive fields. An unspecific receptive field enlargement towards both the stimulated and non-stimulated side was observed in 34% of the tested cells. In contrast to the cells with specifically enlarged receptive fields, the unspecific increase of receptive field size was always accompanied by a strong increase of the general activity level. We conclude that the receptive field changes presumably took place by strengthening of synaptic inputs at the recorded cells in a Hebbian way as previously shown in the visual cortex in vitro and in vivo. The observed receptive field changes may be related to preattentive perceptual learning and could represent a basis of the 'filling in' of cortical scotomas obtained with specific training procedures in human patients suffering from visual cortex lesions.

Influence of GABA-induced remote inactivation on the orientation tuning of cells in area 18 of feline visual cortex: a comparison with area 17.

We have investigated the effect of iontophoretically applying the inhibitory transmitter gamma-aminobutyric acid (GABA) through four pipettes, each located at a horizontal distance of some 500-600 microns from the recording site, on the orientation tuning of cells in areas 17 and 18 of the cat visual cortex for moving the stationary flash-presented bar stimuli. Forty-five of 74 cells tested in area 18 (61%) showed a significant (greater than 25%) increase in orientation tuning width (at half the maximum response) during GABA application, which reflected an increase in response to non-optimal orientations. The mean orientation tuning width of these cells increased by 79%, and the ratio of responses to the orientation orthogonal to the optimum and to the optimum increased from 0.16 to 0.46. The results were similar to those from area 17, in which 36 of 54 cells (66%) showed significant broadening of orientation tuning during GABA application, with a 90% increase in mean tuning width and an increase in the relative response to the orientation orthogonal to the optimum from 0.17 to 0.42. The distributions of cells in areas 17 and 18 with respect to the magnitude of GABA-induced effects on orientation tuning width were not significantly different (mean increase in tuning width: area 17, 102%; area 18, 87%). Although most cells were tested only with moving bars, comparable effects of remote GABA application on orientation tuning were observed when stationary flash-presented bars were used. Of 11 cells thus tested in area 18, seven showed significantly broader tuning during GABA application, with a 132% increase in mean tuning width. In some 25% of cells in each area which showed a significant effect of GABA application on orientation tuning the response to at least one non-optimal orientation exceeded, during GABA application, the response to the previous optimum. There was essentially no correlation between the changes in orientation tuning and changes in the level of spontaneous activity or in the response to the optimum orientation during GABA application. Thus, an increase in the general excitability of recorded cells or the loss of an unspecific inhibitory input cannot account for the effects of GABA application on orientation tuning. Remote GABA application presumably inactivated cells with different preferred orientations from that of the recorded cell. It is thus argued that the observed broadening of orientation tuning during GABA application reflected the loss of an inhibitory input tuned to non-optimal orientations.(ABSTRACT TRUNCATED AT 400 WORDS)

Synaptic transmission in the neocortex during reversible cooling.

We studied the effects of reversible cooling on synaptic transmission in slices of rat visual cortex. Cooling had marked monotonic effects on the temporal properties of synaptic transmission. It increased the latency of excitatory postsynaptic potentials and prolonged their time-course. Effects were non-monotonic on other properties, such as amplitude of excitatory postsynaptic potentials and generation of spikes. The amplitude of excitatory postsynaptic potentials increased, decreased, or remain unchanged while cooling down to about 20 degrees C, but thereafter it declined gradually in all cells studied. The effect of moderate cooling on spike generation was increased excitability, most probably due to the ease with which a depolarized membrane potential could be brought to spike threshold by a sufficiently strong excitatory postsynaptic potential. Stimuli that were subthreshold above 30 degrees C could readily generate spikes at room temperature. Only at well below 10 degrees C could action potentials be completely suppressed. Paired-pulse facilitation was less at lower temperatures, indicating that synaptic dynamics are different at room temperature as compared with physiological temperatures. These results have important implications for extrapolating in vitro data obtained at room temperatures to higher temperatures. The data also emphasize that inactivation by cooling might be a useful tool for studying interactions between brain regions, but the data recorded within the cooled area do not allow reliable conclusions to be drawn about neural operations at normal temperatures.

Effect of partial sensory deprivation on monoaminergic neuromodulators in striate cortex of adult cat.

The role of monoaminergic neuromodulators in the reorganization of cortical topography following limited sensory deprivation in the adult cat was investigated. The total concentrations of dopamine, noradrenaline, serotonin and their major metabolites were measured in the visual cortex of both normal control and experimental animals using microbore high-performance liquid chromatography coupled with electrochemical detection. The experimental animals were subjected to a binocular retinal lesion corresponding to the central 10 degrees of vision and killed two weeks post-lesion. The sensory deprivation was confirmed in area 17 by measuring immediate-early gene zif-268 messenger RNA expression. Following the retinal lesion, the total concentrations of noradrenaline and dopamine were significantly higher in the non-deprived cortex of retinal lesion cats than in the deprived cortex of retinal lesion cats and the cortex of normal animals. This pattern follows the release of the excitatory neurotransmitter glutamate under the same conditions. Serotonin levels were significantly lower in the deprived cortex, and its metabolite 5-hydroxyindole-3-acetic acid was significantly higher in the non-deprived cortex than in deprived cortex and normal cortex. From these results, we suggest that the modulation of noradrenaline, dopamine and serotonin is regulated by visual afferent activity.

Interaction between intracellular tetanization and pairing-induced long-term synaptic plasticity in the rat visual cortex.

Long-term changes in synaptic transmission in slices of rat visual cortex were induced either by pairing the excitatory postsynaptic potentials with postsynaptic depolarization or by intracellular tetanization without synaptic stimulation. Changes in the excitatory postsynaptic potential amplitude induced by any of the protocols applied in isolation persisted for longer than 1 h. Pairing-induced long-term potentiation was input specific. We studied the interaction between intracellular tetanization and pairing-induced plasticity by applying the two protocols one after the other at 10-min intervals. The pairing procedure applied after intracellular tetanization did not lead to any further potentiation, but to a depotentiation of the potentiated inputs. A second pairing protocol applied 10 min later led to further depotentiation, while previously unaffected inputs became weakly depressed. If intracellular tetanization was applied after the pairing procedure, the synaptic responses did not change immediately, but a slow return of the excitatory postsynaptic potential amplitude to the control level could be observed. Therefore, intracellular tetanization is not capable of inducing further potentiation after pairing, and pairing cannot further potentiate the inputs which have already been potentiated by intracellular tetanization. The maintenance of long-term potentiation induced by any of the protocols was impaired by successive application of another procedure. These results suggest a similarity of the mechanisms of synaptic changes induced by the two protocols and demonstrate that the direction of synaptic gain change depends on the history of the synapse.

Retinotopic map plasticity in adult cat visual cortex is accompanied by changes in Ca2+/calmodulin-dependent protein kinase II alpha autophosphorylation.

In adult cats, the induction of homonymous binocular central retinal lesions causes a dramatic reorganization of the topographic map in the sensory-deprived region of the primary visual cortex. To investigate the possible involvement of the alpha-subunit of the calcium/calmodulin dependent protein kinase type II (alphaCaMKII) in this form of brain plasticity, we performed in situ hybridization and Western blotting experiments to analyze mRNA, protein and autophosphorylation levels of this multifunctional kinase. No differences in the mRNA or protein levels were observed between the central, sensory-deprived and the peripheral, non-deprived regions of area 17 of retinal lesion animals or between corresponding cortical regions of normal control animals. Western blotting with an alphaCaMKII threonine-286 phosphorylation-state specific antiserum consistently showed a small, albeit not significant, increase of alphaCaMKII autophosphorylation in the central versus the peripheral region of cortical area 17, and this both in normal subjects as well as in retinal lesion animals with a 3-day post-lesion survival time. In contrast, a post-lesion survival time of 14 days resulted in a alphaCaMKII autophosphorylation level that was four times higher in visually-deprived area 17 than in the non-deprived cortical region. This increased phosphorylation state is not a direct consequence of the decrease in visual activity in these neurons, because we would have expected to see a similar change at shorter or longer post-lesion survival times or in the visually deprived visual cortex of animals in which the left optic tract and the corpus callosum were surgically cut. No such changes were observed, leading to the conclusion that the phosphorylation changes observed at 14 days are related to a delayed reorganization of the retinotopic map of the striate cortex.