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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 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. [ABSTRACT FROM AUTHOR]

The effect of peripheral sensory deprivation upon GABA receptor binding of [H]muscimol was investigated in the barrel cortex - cortical representation of mystacial vibrissae of mice - by means of in vitro quantitative autoradiography. Unilateral lesions of all vibrissae or selected rows of whiskers were performed neonatally or in adulthood. [H]muscimol binding was examined after various survival times up to 60 days. Both types of lesions performed in adult mice resulted in a transient decrease (10-25%) of binding values in the deafferented areas of the barrel field as compared with the unoperated control side. Sixty days after denervation [H]muscimol binding returned to control values. Similar results were found after neonatal removal of all vibrissae. Neonatal lesion of selected rows of vibrissae, however, resulted in a decrease of [H]muscimol binding (by about 26%) lasting up to 60 days in corresponding rows of barrels. This last result was accompanied by severe cytoarchitectonic malformation of the barrel field. The results support the hypothesis that a decrease of inhibition plays a facilitatory role in the plastic reorganization of cortical circuitry. [ABSTRACT FROM AUTHOR]

Abstract Background It has been postulated that exercise-induced activation of brain-derived neurotrophic factor (BDNF) may account for improvement of stepping ability in animals after complete spinal cord transection. As we have shown previously, treadmill locomotor exercise leads to up-regulation of BDNF protein and mRNA in the entire neuronal network of intact spinal cord. The questions arise: (i) how the treadmill locomotor training, supplemented with tail stimulation, affects the expression of molecular correlates of synaptic plasticity in spinal rats, and (ii) if a response is related to BDNF protein level and distribution. We investigated the effect of training in rats spinalized at low thoracic segments on the level and distribution of BDNF immunoreactivity (IR) in ventral quadrants of the lumbar segments, in conjunction with markers of presynaptic terminals, synaptophysin and synaptic zinc. Results Training improved hindlimb stepping in spinal animals evaluated with modified Basso-Beattie-Bresnahan scale. Grades of spinal trained animals ranged between 5 and 11, whereas those of spinal were between 2 and 4. Functional improvement was associated with changes in presynaptic markers and BDNF distribution. Six weeks after transection, synaptophysin IR was reduced by 18% around the large neurons of lamina IX and training elevated its expression by over 30%. The level of synaptic zinc staining in the ventral horn was unaltered, whereas in ventral funiculi it was decreased by 26% postlesion and tended to normalize after the training. Overall BDNF IR levels in the ventral horn, which were higher by 22% postlesion, were unchanged after the training. However, training modified distribution of BDNF in the processes with its predominance in the longer and thicker ones. It also caused selective up-regulation of BDNF in two classes of cells (soma ranging between 100-400 μm2 and over 1000 μm2) of the ventrolateral and laterodorsal motor nuclei. Conclusion Our results show that it is not BDNF deficit that determines lack of functional improvement in spinal animals. They indicate selectivity of up-regulation of BDNF in distinct subpopulations of cells in the motor nuclei which leads to changes of innervation targeting motoneurons, tuned up by locomotor activity as indicated by a region-specific increase of presynaptic markers.

The article presents an abstract of the research paper "NDMA-Induced 45Ca Release in Dentate Gyrus of Newborn rats." The paper explains that the release of 45Ca from the ryanodine-sensitive Ca2+ pool in the dentate gyrus of immature rats participates in generation of NMDA-induced Ca2+ signaling, whereas the role of calbindin in the mechanism of 45Ca release is negligible.

Matrix metalloproteinase (MMP) activity is implicated in the degradation of the extracellular matrix during cerebral ischemia. Although many studies have demonstrated spatiotemporal patterns of activation of gelatinases (MMP-9 and MMP-2) after ischemic stroke in young adult rodents, no data exist on MMP activity in old brains. In this study, we investigated the gelatinolytic activity in young adult (3-month-old) and aged (1-year-old) mice subjected to photothrombotic stroke. Using in situ zymography and gel zymography, we found that the basal gelatinolytic activity in the intact cerebral cortex was similar at both investigated ages. Similarly, after photothrombosis, the increased gelatinolytic response up to 7 days poststroke was the same in young and aged brains. At both ages, early activation of gelatinolysis in the ischemic core and the perilesional area was present in neuronal nuclei as revealed by colocalization of gelatinolytic product with NeuN immunostaining and DAPI. Additionally, application of specific antibodies against MMP-9 and MMP-2 revealed the increase in MMP-9 immunoreactivity in cell nuclei as early as 4 hr poststroke. No differences between young and aged mice were observed concerning the level and localization of MMP-9 immunoreactivity. The lack of age-related differences in the degree and pattern of activation of gelatinolysis after focal stroke and the lack of correspondence between the results of in situ and gel zymography suggest that extracellular proteolysis is not directly responsible for the more severe outcome of ischemic stroke in aged subjects., (Copyright © 2011 Wiley Periodicals, Inc.)

Perineuronal nets (PNNs) are a condensed form of extracellular matrix that covers the surface of a subset of neurons. Their presence limits neuronal plasticity and may protect neurons against harmful agents. Here we analyzed the relationship between spatiotemporal changes in PNN expression and cell death markers after focal cortical photothrombotic stroke in rats. We registered a substantial decrease in PNN density using Wisteria floribunda agglutinin staining and CAT-315 and brevican immunoreactivity; the decrease occurred not only in the lesion core but also in the perilesional and remote cortex as well as in homotopic contralateral cortical regions. Fluoro Jade C and TUNEL staining in perilesional and remote areas, however, showed a low density of dying cells. Our results suggest that the PNN reduction was not a result of cellular death and could be considered an attempt to create conditions favorable for synaptic remodeling., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Neurotransmission is strongly affected after ischaemic insult. It is postulated that modulatory neurotransmitter systems and their receptors play a role in experience-dependent and restoration plasticity. In this study, muscarinic cholinergic, serotonergic 5-HT(2A/2C), dopaminergic D(1) and noradrenergic beta(1) receptors were examined after focal cerebral ischaemia in different brain regions, using quantitative in vitro autoradiography. There were six evaluated time points: 4h, 1, 4, 7, 28 and 60 days after the insult. Rats received unilateral ischaemic lesions through photo-thrombosis in the primary somatosensory cortex. In the lesion core, 5-HT(2A/2C), D(1) and beta(1) receptor binding values return to control levels 28 days after displaying initial decreases, while muscarinic binding remains very low, at 30% of controls. From 4h to 60 days post-stroke no changes are observed in the perilesional tissue. In contrast, in remote brain regions, a bilateral increase of serotonergic 5-HT(2A/2C) receptor binding in the somatosensory cortex at the striatum level is observed after 4h and after 7 days post-stroke. In addition, a bilateral decrease of muscarinic cholinergic receptor binding in the hippocampus is observed at each time point examined. This study points to a complex and remote reaction of modulatory systems in response to ischaemic lesions., (2010 Elsevier B.V. All rights reserved.)

The ability to undergo experience-dependent plasticity in the neocortex is often limited to early development, but also to particular cortical loci and specific experience. In layers II-IV of the barrel cortex, plasticity evoked by removing all but one vibrissae (univibrissa rearing) does not have a time limit except for layer IV barrels, where it can only be induced during the first postnatal week. In contrast, deprivation of every second vibrissa (chessboard deprivation) removes time limits for plasticity. The mechanism permitting plasticity in response to chessboard deprivation and halting it in reply to univibrissa rearing is unknown. Condensation of chondroitin sulfate proteoglycans into perineuronal nets and an increase in intracortical inhibition mediated by parvalbumin-containing interneurons are implicated in closing the critical period for ocular dominance plasticity. These factors could also be involved in setting up the critical period in barrels in a way that depends on a particular sensory experience. We therefore examined changes in density of parvalbumin-containing cells and perineuronal nets during development of mouse barrel cortex and after brief univibrissa and chessboard experience in adolescence. We observed a progressive increase in the density of the two markers across cortical layers between postnatal day 10 and 20, which was especially pronounced in the barrels. Univibrissa rearing, but not chessboard deprivation, increased the density of perineuronal nets and parvalbumin-containing cells in the deprived barrels, but only those that immediately neighbour the undeprived barrel. These data suggest the involvement of both tested factors in closing the critical period in barrels in an experience-dependent manner.

Levels of gamma-aminobutyric acid (GABA) and its synthesizing enzyme in cerebral cortex are regulated by sensory experience. Previously we found that associative pairing of vibrissae stimulation and tail shock results in upregulation of GABAergic markers in the mouse barrel cortex. In order to ascertain whether GABAergic upregulation also accompanies associative pairing in other sensory modalities, we examined the mouse visual cortex after analogous training with visual stimulus. During pairing, visual stimulus (CS) was coupled with a tail shock (UCS). We examined the density of cells expressing glutamic acid decarboxylase (GAD) and parvalbumin (PV) in monocular and binocular segments of the primary visual cortex (V1). The auditory cortex was used as a control. After monocular training, the density of cells expressing GAD rose significantly in the monocular segment of V1 contralateral to the stimulated eye, compared with the opposite hemisphere. This effect was due to the association of CS and UCS, as no changes were found after visual stimulation alone or in the auditory cortex. No changes were noted in the density of PV(+) neurons, so the effect was attributed to GAD(+)/PV(-) neurons. Mobilization of a specific subclass of GABAergic cells, observed after associative pairing in the somatosensory and visual cortices, may reflect the necessity to restrict the activity of circuits involved in sensory association.

Perineuronal nets represent well-organised components of the extracellular matrix, which are surrounding cell bodies, dendrites, and axon segments of a particular class of neurones as well as forming lattice-like structures. The role of perineuronal nets is not fully elucidated yet. Perineuronal nets may play a beneficial role by stabilizing the extracellular milieu assuring the characteristic features of enveloped neurons and protecting them from the influence of harmful agents. On the other hand, perineuronal nets create a barrier which limits neuronal plasticity and counteracts regeneration. This review examines recent evidence concerning the significance of the occurrence of perineuronal nets.

The effects of photothrombotic stroke in primary somatosensory cortex on astroglial and microglial activation in various regions of lesioned brain were examined at different time points, using immunohistochemistry and lectin binding. The increase in GFAP expression was observed exclusively in the ipsilateral hemisphere, both in the perilesional area and cortical regions distant from the infarct. This remote increase was detectable up to sixty days after the infarct. Transient GFAP elevation was also found in the hippocampus one day after photothrombosis, whereas it was more prolonged in amygdala, as demonstrated at four days after lesion. In contrast to a widespread astrocytic activation, the microglial response was shortlasting and local, confined to lesion and perilesional area. Widespread and prolonged activation of astrocytes after stroke may provide factors promoting slowly developing recovery processes in the whole brain, while microglial response seems to be involved in local repair and removal of cellular debris.

Glutamate is the predominant excitatory neurotransmitter in the central nervous system (CNS) and glutamatergic transmission is critical for controlling neuronal activity. Glutamate is stored in synaptic vesicles and released upon stimulation. The homeostasis of glutamatergic system is maintained by a set of transporters present in plasma membrane and in the membrane of synaptic vesicles. The family of vesicular glutamate transporters in mammals is comprised of three highly homologous proteins: VGLUT1-3. The expression of particular VGLUTs is largely complementary with limited overlap and so far they are most specific markers for neurons that use glutamate as neurotransmitter. VGLUTs are regulated developmentally and determine functionally distinct populations of glutamatergic neurons. Controlling the activity of these proteins could potentially modulate the efficiency of excitatory neurotransmission. This review summarizes the recent knowledge concerning molecular and functional characteristic of vesicular glutamate transporters, their development, contribution to synaptic plasticity and their involvement in pathology of the nervous system.

Electroconvulsive seizures (ECS), one of the most effective treatments of depression, induce mossy fiber sprouting (when assayed by means of synaptic zinc method), and this indicates an increase in the synaptic zinc level in the hippocampus following such therapy. The aim of the present study was to investigate the influence of acute and chronic zinc hydroaspartate administration on the synaptic and total zinc level in the rat hippocampus. We used two methods of zinc determination: (1) zinc-selenium method, which images the pool of synaptic zinc, and (2) flame atomic absorption spectrometry, which assays the total concentration of zinc. Our results indicate that chronic (14 x 65 mg/kg), but not acute, zinc hydroaspartate administration intraperitoneally (i.p.) increases the pool of synaptic zinc in the majority of rat hippocampal layers (by 72-190%), except for the stratum moleculare and stratum radiatum CA, and perforant path DG. On the other hand, no changes were found in total hippocampal zinc level, measured by flame atomic absorption spectrometry. These data suggest that chronic zinc treatment increases the pool of synaptic zinc in the hippocampus, and this effect is similar to that observed following chronic ECS treatment. The measurement of zinc concentration in the whole hippocampus by the flame atomic absorption spectrometry method is not sensitive enough to detect such subtle alteration.

In the neocortex, synaptic zinc level is regulated by sensory experience. Previously, we found that trimming of mystacial vibrissae resulted in an increase of synaptic zinc level in corresponding deprived barrels in the cortex of mice. The present study focused on the relationship between synaptic zinc and zinc transporter 3 (ZnT3) protein expression in the barrel cortex of mice during postnatal development and after sensory deprivation of selected vibrissae. Using immunocytochemistry and western blot analysis, we found that ZnT3 expression is delayed as compared with the onset of synaptic zinc and presynaptic markers, such as synapsin I and synaptophysin. Further, neither long-term deprivation in young mice nor short deprivation in adult mice, that resulted in an increase of synaptic zinc level, produced alterations in ZnT3, synapsin I or synaptophysin expression in deprived barrels. These results suggest that in the barrel cortex ZnT3, synapsin I or synaptophysin are not determinant for the activity-dependent regulation of the synaptic zinc level.

The effect of focal photothrombotic stroke on the distribution of D1 dopamine receptor (D1R) sites was examined in different cortical areas of rat brain with quantitative receptor autoradiography using [3H]SCH23390 as a ligand. Unilateral cortical stroke was located in the primary somatosensory cortex. After different survival times (1, 7 and 28 days) D1R binding levels were determined in the lesion core, penumbra, frontoparietal motor (FrPaM) and somatosensory (FrPaSS) areas as well as in homotopic regions in the contralateral hemisphere. One day after stroke, D1R density decreased by 36% (P < 0.01) in the lesion core relative to sham-operated controls. At 7th day binding density was further reduced by 56% (P < 0.002). Twenty-eight days after infarction, D1R binding returned to control level. No alterations in D1R binding levels were found in penumbra and other investigated regions. We suggest that the return of D1R binding to control level in the area initially corresponding to the infarct results from the shrinkage of the lesion volume.

It has been previously demonstrated that in the mouse barrel cortex, synaptic zinc is regulated by sensory experience. In adult mice, cutting selected vibrissae produced a rapid but transient elevation of synaptic zinc in the corresponding barrels several hours later, whereas in 8 day-old animals this procedure did not affect synaptic zinc. In the present study, we wished to determine the postnatal age at which zinc-containing terminals gain the ability to respond rapidly to a restriction of sensory input. We therefore examined the effects of 1-day sensory deprivation starting at different postnatal ages. For this purpose we unilaterally trimmed all rows of vibrissae, except for row C, and we then visualized synaptic zinc in the barrel cortex 24h later. Up to postnatal day 15 such procedure had no effect on the level of synaptic zinc. However, beginning at postnatal day 16, 1-day sensory deprivation produced an increase in synaptic zinc within hollows of deprived rows of barrels as compared to non-deprived rows. These results show that during development there is a specific time-point after which zinc-containing circuits may respond rapidly to altered sensory inputs. A comparison of these findings with previous results obtained after chronic sensory deprivation suggests that a specific time window exists in development for persistent alterations in zinc-containing circuits.

Synapsins are a family of proteins associated with synaptic vesicles that are widely used as markers of synaptic terminals. We decided to investigate synapsin I expression in the mouse primary somatosensory cortex (SI). Immunostaining experiments using a polyclonal antibody against C-terminal domain of synapsin Ia/b (anti-SynI-C) showed an unusual pattern in the SI cortex compared to other regions of the neocortex. The staining delineated the cells located in barrel hollows. The immunoreactive product was located on the perikarya and proximal dendrites of gabaergic neurons found in layer IV and VI of the SI cortex. Other anti-synapsin antibodies did not reveal this pattern within the SI cortex, although in the hippocampus all antibodies examined produced a similar pattern of immunostaining. Deglycosylation of sections resulted in complete loss of immunodecoration on the cell perikarya. We suggest that the anti-SynI-C recognizes a saccharide surface epitope, possibly an element of perineuronal nets that is specific for the primary somatosensory cortex.

The distribution of synaptic zinc after short-term (up to 48 h) tactile deprivation of vibrissae was investigated in the barrel cortex of mice using histochemical staining. In adult mice, 12 h after trimming selected rows of vibrissae, an increase in zinc staining in the deprived barrels was observed. This increase was still present 48 h after trimming. These results indicate that the level of synaptic zinc is rapidly regulated by neuronal activity in adult mice. In young (8-day-old) mice, the short-term deprivation did not alter zinc staining and only chronic sensory deprivation produced an increase in zinc staining. However, after long-term deprivation no changes were found in adult mice. These results suggest that different mechanisms might be involved in functional reorganization of zinc containing terminals in young and fully mature cerebral 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., (Copyright 1999 Elsevier Science B.V.)

Histochemical localization of synaptic zinc was examined in the somatosensory (SI) barrel cortex of mouse. The laminar distribution and distribution within the barrel field were described. At postnatal day 3 (P3) and 5 (P5), very faint and uniform zinc staining was present in the lower part of the subplate. At P6, subtle laminar variations emerged. At P8, these variations were clearly observed. Intense zinc staining was found in layers I, II, III, and V. Layers IV and VI showed a weaker staining. From this postnatal age to adult, uneven patchy distribution of synaptic zinc in layer IV could be distinguished in coronal sections. In tangential sections through layer IV, zinc staining showed a barrel-like pattern due to a higher zinc concentration in septa and the surrounding cortex. Barrel sides revealed a lower zinc concentration compared with the barrel hollow. With brain maturation, the zinc staining increased more intensely outside the barrel field, thus producing a progressively higher contrast between the barrel field and adjacent cortical regions. The differences in zinc staining between the barrel side and barrel hollow diminished with age but were still visible at P70. The changes in synaptic zinc distribution probably reflect the process of synaptic maturation of glutamatergic terminals projecting to the SI cortex. The time course of postnatal changes in terminal zinc distribution suggests that synaptic zinc is not involved in the mechanisms of barrel formation.

The effects of sensory stimulation and sensory conditioning upon [3H]MK-801 [(+)-5-methyl-10,11-dihydro- 5H-dibenzo[a,d]cyclohepten-5, 10-imine] binding to N-methyl- d-aspartate (NMDA) receptor sites and [3H]AMPA (alpha-amino-3-hydroxy-5-methylisoxasole-4-propionic acid) binding to AMPA receptor sites were examined in the primary somatosensory (SI) cortex of mice. Following short-lasting unilateral tactile stimulation of a selected row of vibrissae, and tactile stimulation paired with noxious stimulus (the pairing procedure was found to alter cortical representation of vibrissae), in vitro receptor binding autoradiography was performed on the sections of the barrel cortex of mice. One hour after the end of tactile stimulation or training procedure there was an increase of [3H]MK-801 and [3H]AMPA binding in the corresponding row of barrels in layer IV of the SI cortex of adult mice. These effects disappeared 24 h after the end of each experimental procedure. The results suggest that both subtypes of glutamate receptors are regulated in an activity-dependent way and that sensory stimulation transiently modifies local cortical processing.