Impact of somatostatin interneurons on interactions between barrels in plasticity induced by whisker deprivation.

The activity of inhibitory interneurons has a profound role in shaping cortical plasticity. Somatostatin-expressing interneurons (SOM-INs) are involved in several aspects of experience-dependent cortical rewiring. We addressed the question of the barrel cortex SOM-IN engagement in plasticity formation induced by sensory deprivation in adult mice (2-3 months old). We used a spared vibrissa paradigm, resulting in a massive sensory map reorganization. Using chemogenetic manipulation, the activity of barrel cortex SOM-INs was blocked or activated by continuous clozapine N-oxide (CNO) administration during one-week-long deprivation. To visualize the deprivation-induced plasticity, [14C]-2-deoxyglucose mapping of cortical functional representation of the spared whisker was performed at the end of the deprivation. The plasticity was manifested as an extension of cortical activation in response to spared vibrissae stimulation. We found that SOM-IN inhibition in the cortical column of the spared whisker did not influence the areal extent of the cortex activated by the spared whisker. However, blocking the activity of SOM-INs in the deprived column, adjacent to the spared one, decreased the plasticity of the spared whisker representation. SOM-IN activation did not affect plasticity. These data show that SOM-IN activity is part of cortical circuitry that affects interbarrel interactions underlying deprivation-induced plasticity in adult mice.

Learning-induced plasticity in the barrel cortex is disrupted by inhibition of layer 4 somatostatin-containing interneurons.

Gaba-ergic neurons are a diverse cell class with extensive influence over cortical processing, but their role in experience-dependent plasticity is not completely understood. Here we addressed the role of cortical somatostatin- (SOM-INs) and vasoactive intestinal polypeptide- (VIP-INs) containing interneurons in a Pavlovian conditioning where stimulation of the vibrissae is used as a conditioned stimulus and tail shock as unconditioned one. This procedure induces a plastic change observed as an enlargement of the cortical functional representation of vibrissae activated during conditioning. Using layer-targeted, cell-selective DREADD transductions, we examined the involvement of SOM-INs and VIP-INs activity in learning-related plastic changes. Under optical recordings, we injected DREADD-expressing vectors into layer IV (L4) barrels or layer II/III (L2/3) areas corresponding to the activated vibrissae. The activity of the interneurons was modulated during all conditioning sessions, and functional 2-deoxyglucose (2DG) maps were obtained 24 h after the last session. In mice with L4 but not L2/3 SOM-INs suppressed during conditioning, the plastic change of whisker representation was absent. The behavioral effect of conditioning was disturbed. Both L4 SOM-INs excitation and L2/3 VIP-INs inhibition during conditioning did not affect the plasticity or the conditioned response. We found the activity of L4 SOM-INs is indispensable in the formation of learning-induced plastic change. We propose that L4 SOM-INs may provide disinhibition by blocking L4 parvalbumin interneurons, allowing a flow of information into upper cortical layers during learning.

Interneurons containing somatostatin are affected by learning-induced cortical plasticity.

The maintenance of neural circuit stability is a dynamic process that requires the plasticity of many cellular and synaptic components. By changing the excitatory/inhibitory balance, inhibitory GABAergic plasticity can regulate excitability, and contribute to neural circuit function and refinement in learning and memory. Increased inhibitory GABAergic neurotransmission has been shown in brain structures involved in the learning process. Previously, we showed that classical conditioning in which tactile stimulation of one row of vibrissae (conditioned stimulus, CS) was paired with a tail shock (unconditioned stimulus, UCS) in adult mice results in the increased density of GABAergic interneurons and increased expression of glutamic acid decarboxylase (GAD)-67 in barrels of the "trained" row cortical representation. In inhibitory neurons of the rat cortex GAD co-localizes with several proteins and peptides. We found previously that the density of the parvalbumin (GAD+/Prv+)-containing subpopulation is not changed after conditioning. In the present study, we examined GABAergic somatostatin (Som)-, calbindin (CB)- and calretinin (CR)-positive interneurons in the cortical representation of "trained" vibrissae after training. Cells showing double immunostaining for GAD/Som, GAD/CR and GAD/CB were counted in the barrels representing vibrissae activated during the training and in control, untouched rows. We found a substantial increase of GAD/Som-containing cells in the trained row representation. No changes in the density of GAD/CR or GAD/CB neurons were observed. These results suggest that Som-containing interneurons are involved in learning-induced changes in the inhibitory cortical network.

Correlated activation of the thalamocortical network in a simple learning paradigm.

The thalamocortical loop is a key player in sensory processing. We examined the functional interactions among its elements, expressed as cross-correlations between metabolic activity of the barrel cortex, somatosensory thalamic nuclei and posterior parietal cortex, in classical conditioning. In the training stimulation of vibrissae in mice was paired with a tail shock. [14C]-2-Deoxyglucose brain mapping was performed during the first and the final sessions of conditioning (conditioned stimulus+unconditioned stimulus; CS+UCS), in groups that received only the stimulation of vibrissae (conditioned stimulus; CS-only) and in nonstimulated controls (NS). In the CS-only group, the CS evoked the correlated activity of the examined structures during the first session, but in the third session these structures did not act in a correlated manner. Conversely, in the CS+UCS condition correlations among the thalamocortical loop structures activities became stronger during the course of the training. Particularly, the posterior parietal cortex, which controls voluntary deployment of attention, together with the barrel cortex becomes involved in the network of structures with the correlated activity. The results suggest a predominant role for bottom-up processing in the somatosensory pathway at the beginning of conditioning followed by top-down processing. This is consistent with the idea that the thalamocortical loop plays a crucial role in attentional processes.

Emotionally negative stimuli can overcome attentional deficits in patients with visuo-spatial hemineglect.

Left unilateral spatial neglect resulting from right brain damage is characterized by loss of awareness for stimuli in the contralesional side of space, despite intact visual pathways. We examined using fMRI whether patients with neglect are more likely to consciously detect in the neglected hemifield, emotionally negative complex scenes rather than visually similar neutral pictures and if so, what neural mechanisms mediate this effect. Photographs of emotional and neutral scenes taken from the IAPS were presented in a divided visual field paradigm. As expected, the detection rate for emotional stimuli presented in the neglected field was higher than for neutral ones. Successful detection of emotional scenes as opposed to neutral stimuli in the left visual field (LVF) produced activations in the parahippocampal and anterior cingulate areas in the right hemisphere. Detection of emotional stimuli presented in the intact right visual field (RVF) activated a distributed network of structures in the left hemisphere, including anterior and posterior cingulate cortex, insula, as well as visual striate and extrastriate areas. LVF-RVF contrasts for emotional stimuli revealed activations in right and left attention related prefrontal areas whereas RVF-LVF comparison showed activations in the posterior cingulate and extrastriate visual cortex in the left hemisphere. An additional analysis contrasting detected vs. undetected emotional LVF stimuli showed involvement of left anterior cingulate, right frontal and extrastriate areas. We hypothesize that beneficial role of emotion in overcoming neglect is achieved by activation of frontal and limbic lobe networks, which provide a privileged access of emotional stimuli to attention by top-down modulation of processing in the higher-order extrastriate visual areas. Our results point to the importance of top-down regulatory role of the frontal attentional systems, which might enhance visual activations and lead to greater salience of emotional stimuli for perceptual awareness.

Matrix metalloproteinase inhibition counteracts impairment of cortical experience-dependent plasticity after photothrombotic stroke.

Matrix metalloproteinases (MMPs) are fine modulators of brain plasticity and pathophysiology. The inhibition of MMPs shortly after ischaemic stroke reduces the infarct size and has beneficial effects on post-stroke behavioural recovery. Our previous studies have shown that photothrombotic cortical stroke disrupts use-dependent plasticity in the neighbouring cortex. The aim of the present study was to check whether the inhibition of MMPs after photothrombosis rescued the plastic capacity of the barrel cortex. To induce plasticity in adult mice, a unilateral deprivation of all vibrissae except row C was applied. The deprivation started immediately after stroke and lasted 7 days. This procedure, in control (non-stroke) animals, results in an enlargement of functional representation of the spared row, as shown with [(14)C]2-deoxyglucose uptake mapping. In mice with stroke induced by photothrombosis in the vicinity of the barrel cortex, vibrissae deprivation did not result in an enlargement of the cortical representation of the spared row C of vibrissae, which confirmed our previous results. However, when mice were injected with the broad-spectrum inhibitor of MMPs FN-439 (10 mg/kg, i.v.) immediately before a stroke, an enlargement of the representation of the spared row similar to the enlargement found in sham mice was observed. These results indicate the involvement of MMPs in the impairment of use-dependent plasticity in the vicinity of an ischaemic lesion.

Remapping of the somatosensory cortex after a photothrombotic stroke: dynamics of the compensatory reorganization.

Although stroke can affect cerebral structure and function, the brain has a potential for plasticity thanks to which some degree of function can be restored. The pathways of such recovery are of great interest, since the dynamics of rewiring of the injured brain may become the basis for designing appropriate strategies of rehabilitation. We investigated the spontaneous plasticity of cortical somatosensory representations following a focal unilateral stroke in the barrel cortex of rats. Ischemic lesions were produced with the photothrombotic technique in the cortical representation of vibrissae. Functional activation of the brain in response to the stimulation of vibrissae with destroyed cortical representation was monitored through the 2 months post-stroke survival period with [(14)C] 2-deoxyglucose (2DG) autoradiographic brain mapping (1, 7, 28, 56 days after the stroke). 2DG uptake was measured on autoradiograms of tangential sections in several regions of somatosensory cortex and in motor, auditory and prefrontal cortex. Behavioral deficit was assessed by the gap-crossing test 3, 28, 56 days after the stroke. Changes in the activation pattern of the intact hemisphere and non-vibrissal somatosensory representations of the lesioned hemisphere evolved during the observation period. Full recovery of the behavioral function was reached 2 months after the stroke and at the same time, new foci of activation were observed in the lesioned hemisphere. At that time, hyperactivation of the somatosensory areas in the intact hemisphere subsided. The new activation foci located in representations of anterior vibrissae, front paw and hind paw were specific for the vibrissae stimulation and were most probably a new functional representation of the vibrissae. We demonstrated spatial and temporal remodeling of the brain induced by cortical stroke, leading to vicariation of function.

Alpha calcium/calmodulin dependent protein kinase II in learning-dependent plasticity of mouse somatosensory cortex.

Calcium/calmodulin dependent protein kinase II (CaMKII), and more specifically its alpha subunit, is widely believed to be fundamental for hippocampal synaptic plasticity. In the cerebral cortex, deprivation-evoked plasticity was shown to depend on alphaCaMKII autophosphorylation abilities. Here we analyzed how learning-induced functional reorganization of cortical representations affected alphaCaMKII in adult Swiss mice. Mice were subjected to short-lasting sensory training in which stimulation of whiskers was paired with tail shock. The pairing results in enlargement of functional representation of vibrissae activated during the training. alphaCaMKII protein and its autophosphorylation level were determined by Western-blotting in somatosensory cortex crude synaptosomal fraction (P2) and postsynaptic protein-enriched, Triton X-100 insoluble fraction (TIF). The first training session resulted in an increase in alphaCaMKII autophosphorylation at autonomy site observed in TIF. A similar increase was also observed after the first session of just whiskers stimulation, which alone does not induce rearrangement of cortical representations. These data indicate that increased autophosphorylation of postsynaptic alphaCaMKII is not a correlate of induction phase of plasticity related reorganization of cortical representation of vibrissae. The increase observed in both experimental groups was transient and did not persist in the maintenance phase of the plastic change. Furthermore, we found that the training caused a delayed upregulation of alphaCaMKII protein level in crude synaptosomal fraction, but not in TIF, and the upregulation was not accompanied by an increase in autophosphorylation level of the kinase. The result indicates alphaCaMKII involvement in the late phase of plastic change and suggests the participation of a presynaptic pool of kinase rather than postsynaptic at this point.

Short-term sensory learning does not alter parvalbumin neurons in the barrel cortex of adult mice: a double-labeling study.

We have previously reported that a classical conditioning paradigm involving stimulation of a row of facial vibrissae produced expansion of the cortical representation of the activated vibrissae ("trained row"), this was demonstrated by labeling with 2-deoxyglucose in layer IV of the barrel cortex. We have also shown that functional reorganization of the primary somatosensory cortex is accompanied by an increase in the density of small GABAergic cells and glutamate decarboxylase 67-positive neurons in the hollows of barrels representing the "trained row." GABA neurons of the rat neocortex co-localize with calcium-binding proteins [parvalbumin, carletinin, calbindin D28k] and neuropeptides (vasoactive intestinal polypeptide, somatostatin). In the present study we have examined GABAergic parvalbumin-positive, interneurons in the cortical representation of "trained" facial vibrissae after short-term aversive training, in order to determine whether the observed changes in glutamate decarboxylase 67-positive neurons are accompanied by changes in parvalbumin-positive neurons. Using double immunofluorescent staining, it was found that (i) all parvalbumin-positive neurons in the barrel hollows were glutamate decarboxylase 67-positive, (ii) following aversive training density of glutamate decarboxylase 67-positive neurons in barrel hollows increased significantly compared with controls and (iii) density glutamate decarboxylase 67-positive/parvalbumin-positive neurons in "trained" barrel hollows did not change compared with controls. This study is the first to demonstrate that the density of double-labeled glutamate decarboxylase 67-positive/parvalbumin-positive neurons does not alter during cortical plasticity, thus suggesting that some other population (i.e. parvalbumin negative) of GABAergic interneurons is involved in learning-dependent changes in layer IV of the barrel cortex.

Experience-dependent changes in cortical whisker representation in the adult mouse: a 2-deoxyglucose study.

Sensory experience and learning can modify cortical body maps. We have previously reported that 3 days of classical conditioning, in which stimulation of a row of whiskers was paired with tail shock, produced an expansion of the cortical representation of the "trained row" labeled with 2-deoxyglucose (2DG), in layer IIIb and IV of the barrel cortex. The present study examined plastic remodelling of the vibrissal cortical representation after pairing whisker stimulation with a drop of sweet water. Cortical representations of rows of whiskers were mapped by 2DG autoradiography after 3 days and 2 months of training. The training resulted in enlargement of the cortical representation of vibrissae involved in the stimulus pairing compared with the contralateral representation of a row of whiskers, that were not touched during the training. This modification of whisker representation was different after short-term and long-term appetitive training. After three pairing sessions, changes in the width of cortical representation were visible in layers II/IIIa (29%) and layers V/VI (28%). After 2 months of training, significant changes in the width of cortical representation row B were found only in layer IV (41%). The changes were not observed in animals, that received whisker stimulation alone or in those who were subjected to training with unpaired stimuli. The results demonstrate that stimulus-pairing-induced changes in cortical whisker representation appeared with different time courses at different levels of cortical columnar information processing.

Delayed upregulation of GABA(A) alpha1 receptor subunit mRNA in somatosensory cortex of mice following learning-dependent plasticity of cortical representations.

Experience-dependent modifications of cortical representational maps are accompanied by changes in several components of GABAergic inhibitory neurotransmission system. We examined with in situ hybridization to 35S-labeled oligoprobe changes of expression of GABA(A) receptor alpha1 subunit mRNA in the barrel cortex of mice after sensory conditioning training. One day and 5 days after the end of short lasting (3 daily sessions) training an increased expression of GABA(A) alpha1 mRNA was observed at the cortical site where the plastic changes were previously found. Learning associated activation of the cerebral cortex increases expression of GABA(A) receptor mRNA after a short post-training delays.

PSD95 protein level rises in murine somatosensory cortex after sensory training.

Proteins of the postsynaptic density are implicated in mechanisms of synaptic plasticity. We examined involvement of PSD95 and alphaCaMkII in learning-dependent plastic changes of representational maps in somatosensory cortex of mice. The barrel cortex of mice was examined following a 3 day long classical conditioning training, in which activation of facial vibrissae was linked to an aversive stimulus. This procedure produced expansion of cortical representations of vibrissae involved in the training. In subcellular fraction enriched in postsynaptic densities from the barrel cortex, it was estimated by Western blotting that the level of PSD95 increased after the training by about 50%, while the level of CaMkII remained unchanged. The results indicate involvement of PSD95 in learning-dependent cortical plasticity.

Rapid regulation of GAD67 mRNA and protein level in cortical neurons after sensory learning.

Cortical representations of different modalities can be modified by sensory learning. Our previous studies in the barrel cortex showed that expansion of the cortical representation of a row of vibrissae could be induced by pairing stimulation of a row of vibrissae with a tail shock. The plastic change in cortical reactivity to the input used during the training was accompanied by increased density of GABA immunoreactive neurons in the involved row of cortical barrels. Using the same paradigm, the present study examined the pathway of GABA synthesis-expression of GAD67 mRNA and immunoreactivity of GAD67 isoenzyme in the barrel cortex of mice after sensory learning. In situ hybridization revealed that the GAD67 mRNA level was elevated in one row of barrels in the trained group as well as in controls receiving vibrissae stimulation alone. In contrast, elevation of immunoreactivity of the GAD67 protein occurred only in the trained group. The density of GABA-immunoreactive neurons in the hollows of barrels representing the row of vibrissae activated during the training was increased by 50%. These data indicated that sensory stimulation alone affected expression of the 67 kDa glutamate decarboxylase isoenzyme synthesis pathway, whereas the processes involved in cortical plasticity induced by associative learning modified this pathway additionally at the level of translation.

Mice can learn roughness discrimination with vibrissae in a jump stand apparatus.

An adaptation of roughness discrimination task successfully used on rats was performed on mice. It was found that mice can master discrimination of rough surfaces using only mystacial vibrissae. This task can be used for studying sensory abilities of genetically modified mice as well as dynamics and pharmacology of complex sensory learning.

Age-dependent response of the mouse barrel cortex to sensory deprivation: a 2-deoxyglucose study.

The effect of age on the plastic response of vibrissal barrel cortex to deprivation was examined in adolescent (1 month at the start of the procedure, 2 months at testing) and mature (10- to 11-month-old) mice. A single vibrissa was plucked out for 3 weeks and allowed to regrow for 10 days; it was previously found that this deprivation paradigm induces strong downregulation of the deprived input. The results of deprivation were assessed with 2-deoxyglucose functional brain-mapping autoradiography. Deprivation was found to reduce the ability of the deprived vibrissa to activate the cortex both in adolescent and mature mice. However, while in young animals the decrease of the extent of cortical labeling, compared with the normal control, was observed in all examined cortical layers (II/III, IV, and V), in older mice the effect was reduced in layers II/III and absent in layer IV. The suppression of response of the infragranular layers was not affected by age. Transition from adolescent to mature adulthood brings about a layer-specific decline in depression of the cortical response to the deprived input.

[Brain plasticity]. / Plastycznosc mózgu.

The ability to undergo lasting changes of neuronal response properties i.e. plasticity, is one of the most important aspects of functioning of the nervous system. The paper reviews the plastic changes of adult mammalian cerebral cortex. It describes compensatory plasticity induced by peripheral denervation and compensatory plasticity following central trauma. Neuronal mechanisms of cortical neuroplastic changes are presented and the common rules of all forms of cortical modifications are discussed. The paper stresses the therapeutic value of training in various sensory modalities for corrections of functional maladaptation of cortical maps and compensation of function after stroke.

GABA immunoreactivity in mouse barrel field after aversive and appetitive classical conditioning training involving facial vibrissae.

We have previously reported that a classical conditioning paradigm involving stimulation of a row of facial vibrissae produced an expansion of the cortical representation of the "trained row", labeled with 2-deoxyglucose (2DG), in layer IV of the barrel field. The present study has examined the pattern of GABA immunoreactivity (GABA-IR) in the cortical representation of row B of the facial vibrissae after (i) 3 days of aversive training, and (ii) 2 months of appetitive training, where stimulation of row B of vibrissae on one side of the snout was used as a conditioned stimulus. The most notable observation was a greater density of GABA-IR cells concentrated in the hollows of the "trained row" B barrels compared to the hollows in the barrel field of the opposite hemisphere in the same mouse. After aversive training, we noted a 2-fold increase in the density of GABA-IR neurons in the hollows of row B; after reward training, the increase amounted to 49%. In contrast, GABA-IR was unchanged in the control groups, which received only stimulation of vibrissae without the unconditioned stimulus. The classification of labeled neurons according to size revealed that the increase in density of GABA-IR neurons was confined to the small (12-15 microm) diameter group. We concluded that the GABAergic system undergoes up-regulation, after both associative learning paradigms, and that the population of small, GABAergic neurons plays an active role in use-dependent plasticity.

Deafferentation induced changes in GAD67 and GluR2 mRNA expression in mouse somatosensory cortex.

Partial vibrissectomy in adult mice induces body map plasticity in SI barrel cortex. To examine if the disturbed balance of cortical activation affects the excitatory and inhibitory neurotransmitter systems, we studied glutamic acid decarboxylase (GAD 67) and AMPA receptor subunit GluR2 mRNA expression in the barrel cortex. At varying times post-vibrissectomy, sparing row C of whiskers on one side of the snout, the brains were processed for in situ hybridization using specific [(35)S]oligonucleotides to detect the laminar localization of GAD67 and GluR2 mRNAs. Three and seven days after vibrissectomy, the expression of GAD67 was decreased in the deafferented cortex, while 30 days post-lesion, no effects were observed. At 3 days post-lesion, an ipsilateral decrease in GAD67 mRNA expression was also observed. No decreases in GluR2 transcripts were found in the deafferented cortex, but an increased expression was observed in the representation of the spared row C of whiskers 3 days after vibrissectomy. Seven and 30 days post lesion no changes in GluR2 expression were found. These data indicate that in the barrel cortex, peripheral deafferentation transiently regulates GAD67 and GluR2 expression at the transcriptional level. We suggest that this may be a manifestation of adaptive processes.

Anatomical correlates of representational map reorganization induced by partial vibrissectomy in the barrel cortex of adult mice.

We examined the potential for changes in cortical connectivity to accompany long-term plastic changes in functional cortical representations of mystacial vibrissae. Plasticity in the barrel cortex of young adult mice was evoked by vibrissectomy that spared row C of whiskers. We found that 2-deoxyglucose brain mapping causes a progressive expansion of cortical representation of the spared vibrissae. Two months after vibrissectomy, when the width of the cortical map of the spared row of vibrissae doubled, living cortical slices of the barrel cortex were injected with fluorescent dextrans. The injections were centered on spared, deprived and control vibrissal columns. The injections labeled three intracortical projection systems: (i) local connections from one vibrissal column to neighboring columns; (ii) long-range projections running in the septa and walls of the barrels and spanning several barrels; and (iii) very-long-range fibers running horizontally in the lower part of layer V. The local, short-range projection system was analysed following small injections into the centers of columns in layers III and IV. We found that injections into spared barrels labeled axons extending for significantly greater distances in all layers (except layer V), and labeled cell bodies situated significantly further, than after injections into deprived or control barrels. Also, the total axonal density labeled by injections into the spared barrel was higher by 70% than for the deprived or control barrels. Alterations of topographical maps in adult somatosensory cortex may occur immediately after functional denervation, but may also increase with time, as in the case of our experimental situation. Our results indicate that persistent, long-term plastic change can remodel connectivity in the barrel cortex.

Partial blocking of NMDA receptors reduces plastic changes induced by short-lasting classical conditioning in the SI barrel cortex of adult mice.

The effect of blockade of N-methyl-D-aspartate (NMDA) receptors in the barrel cortex upon the learning-induced changes of the cortical body map was examined in adult mice. We have previously found that three sensory conditioning sessions, in which stimulation of a row of vibrissae was paired with a tail shock, produced an enlargement of the functional representation of a row of vibrissae stimulated during training. Implantation of the slow release polymer Elvax, containing 2-amino-5-phosphonovalerate (APV, 50 mM), in the vicinity of the barrel cortex was performed 1 day before conditioning to block NMDA receptors. The cortical representation of a trained row of vibrissae was visualized with 2-deoxyglucose (2DG) functional brain mapping 1 day after the completion of the conditioning procedure. The partial blockade of NMDA receptors within the barrel cortex reduced (by half) the expansion of the cortical representation of a trained row of vibrissae as compared to the enlargement of the cortical representation of a trained row found in untreated (60%) and Elvax-PBS implanted (47%) mice. The results provide evidence that the learning-induced processes of cortical map reorganization involve mechanisms that depend on NMDA receptor activation.