Lhx2 regulates a cortex-specific mechanism for barrel formation.

LIM homeodomain transcription factors are critical regulators of early development in multiple systems but have yet to be examined for a role in circuit formation. The LIM homeobox gene Lhx2 is expressed in cortical progenitors during development and also in the superficial layers of the neocortex in maturity. However, analysis of Lhx2 function at later stages of cortical development has been hampered by severe phenotypes associated with early loss of function. We identified a particular Cre-recombinase line that acts in the cortical primordium after its specification is complete, permitting an analysis of Lhx2 function in neocortical lamination, regionalization, and circuit formation by selective elimination of Lhx2 in the dorsal telencephalon. We report a profound disruption of cortical neuroanatomical and molecular features upon loss of Lhx2 in the cortex from embryonic day 11.5. A unique feature of cortical circuitry, the somatosensory barrels, is undetectable, and molecular patterning of cortical regions appears disrupted. Surprisingly, thalamocortical afferents innervate the mutant cortex with apparently normal regional specificity. Electrophysiological recordings reveal a loss of responses evoked by stimulation of individual whiskers, but responses to simultaneous stimulation of multiple whiskers were present, suggesting that thalamic afferents are unable to organize the neurocircuitry for barrel formation because of a cortex-specific requirement of Lhx2. We report that Lhx2 is required for the expression of transcription factor paired box gene 6, axon guidance molecule Ephrin A5, and the receptor NMDA receptor 1. These genes may mediate Lhx2 function in the formation of specialized neurocircuitry necessary for neocortical function.

Ocimum sanctum [Tulsi] as a Potential Immunomodulator for the Treatment of Ischemic Injury in the Brain.

Stroke causes brain damage and is one of the main reasons for death. Most survivors of stroke face long-term physical disabilities and cognitive dysfunctions. In addition, they also have persistent emotional and behavioral changes. The two main treatments that are effective are reperfusion with recombinant tissue plasminogen activator and recanalization of penumbra using mechanical thrombectomy. However, these treatments are suitable only for a few patients due to limitations such as susceptibility to hemorrhage and the requirement for administering tissue plasminogen activators within the short therapeutic window during the early hours following a stroke. The paucity of interventions and treatments could be because of the multiple pathological mechanisms induced in the brain by stroke. The ongoing immune response following stroke has been attributed to the worsening brain injury. Hence, novel compounds with immunomodulatory properties that could improve the outcome of stroke patients are required. Natural compounds and medicinal herbs with anti-inflammatory activities and having minimal or no adverse systemic effect could be beneficial in treating stroke. Ocimum sanctum is a medicinal herb that can be considered an effective therapeutic option for ischemic brain injury. Ocimum sanctum, commonly known as holy basil or "Tulsi," is mentioned as the "Elixir of Life" for its healing powers. Since antiquity, Tulsi has been used in the Ayurvedic and Siddha medical systems to treat several diseases; it possesses immuno-modulatory activity, which can alter cellular and humoral immune responses. Tulsi can be considered a potential option as an immuno-modulator for treating various diseases, including brain stroke. In this review, we will focus on the immunomodulatory properties of Tulsi, specifically its effect on both innate and adaptive immunity, as well as its antioxidant and anti-inflammatory properties, which could potentially be effective in treating ongoing immune reactions following ischemic brain injury.

Cytidine-5-diphosphocholine supplement in early life induces stable increase in dendritic complexity of neurons in the somatosensory cortex of adult rats.

Cytidine-5-diphosphocholine (CDP-choline or citicholine) is an essential molecule that is required for biosynthesis of cell membranes. In adult humans it is used as a memory-enhancing drug for treatment of age-related dementia and cerebrovascular conditions. However the effect of CDP-choline on perinatal brain is not known. We administered CDP-choline to Long Evans rats each day from conception (maternal ingestion) to postnatal day 60 (P60). Pyramidal neurons from supragranular layers 2/3, granular layer 4 and infragranular layer 5 of somatosensory cortex were examined with Golgi-Cox staining at P240. CDP-choline treatment significantly increased length and branch points of apical and basal dendrites. Sholl analysis shows that the complexity of apical and basal dendrites of neurons is maximal in layers 2/3 and layer 5. In layer 4 significant increases were seen in basilar dendritic arborization. CDP-choline did not increase the number of primary basal dendrites on neurons in the somatosensory cortex. Primary cultures from somatosensory cortex were treated with CDP-choline to test its effect on neuronal survival. CDP-choline treatment neither enhanced the survival of neurons in culture nor increased the number of neurites. However significant increases in neurite length, branch points and total area occupied by the neurons were observed. We conclude that exogenous supplementation of CDP-choline during development causes stable changes in neuronal morphology. Significant increase in dendritic growth and branching of pyramidal neurons from the somatosensory cortex resulted in enlarging the surface area occupied by the neurons which we speculate will augment processing of sensory information.

Deficits in Behavioral Functions of Intact Barrel Cortex Following Lesions of Homotopic Contralateral Cortex.

Focal unilateral injuries to the somatosensory whisker barrel cortex have been shown cause long-lasting deficits in the activity and experience-dependent plasticity of neurons in the intact contralateral barrel cortex. However, the long-term effect of these deficits on behavioral functions of the intact contralesional cortex is not clear. In this study, we used the "Gap-crossing task" a barrel cortex-dependent, whisker-sensitive, tactile behavior to test the hypothesis that unilateral lesions of the somatosensory cortex would affect behavioral functions of the intact somatosensory cortex and degrade the execution of a bilaterally learnt behavior. Adult rats were trained to perform the Gap-crossing task using whiskers on both sides of the face. The barrel cortex was then lesioned unilaterally by subpial aspiration. As observed in other studies, when rats used whiskers that directly projected to the lesioned hemisphere the performance of Gap-crossing was drastically compromised, perhaps due to direct effect of lesion. Significant and persistent deficits were present when the lesioned rats performed Gap-crossing task using whiskers that projected to the intact cortex. The deficits were specific to performance of the task at the highest levels of sensitivity. Comparable deficits were seen when normal, bilaterally trained, rats performed the Gap-crossing task with only the whiskers on one side of the face or when they used only two rows of whiskers (D row and E row) intact on both side of the face. These findings indicate that the prolonged impairment in execution of the learnt task by rats with unilateral lesions of somatosensory cortex could be because sensory inputs from one set of whiskers to the intact cortex is insufficient to provide adequate sensory information at higher thresholds of detection. Our data suggest that optimal performance of somatosensory behavior requires dynamic activity-driven interhemispheric interactions from the entire somatosensory inputs between homotopic areas of the cerebral cortex. These results imply that focal unilateral cortical injuries, including those in humans, are likely to have widespread bilateral effects on information processing including in intact areas of the cortex.

Short exposure to an enriched environment accelerates plasticity in the barrel cortex of adult rats.

Cortical sensory neurons adapt their response properties to use and disuse of peripheral receptors in their receptive field. Changes in synaptic strength can be generated in cortex by simply altering the balance of input activity, so that a persistent bias in activity levels modifies cortical receptive field properties. Such activity-dependent plasticity in cortical cell responses occurs in rat cortex when all but two whiskers are trimmed for a period of time at any age. The up-regulation of evoked responses to the intact whiskers is first seen within 24 h in the supragranular layers [Laminar comparison of somatosensory cortical plasticity. Science 265(5180):1885-1888] and continues until a new stable state is achieved [Experience-dependent plasticity in adult rat barrel cortex. Proc Natl Acad Sci U S A 90(5):2082-2086; Armstrong-James M, Diamond ME, Ebner FF (1994) An innocuous bias in whisker use in adult rat modifies receptive fields of barrel cortex neurons. J Neurosci 14:6978-6991]. These and many other results suggest that activity-dependent changes in cortical cell responses have an accumulation threshold that can be achieved more quickly by increasing the spike rate arising from the active region of the receptive field. Here we test the hypothesis that the rate of neuronal response change can be accelerated by placing the animals in a high activity environment after whisker trimming. Test stimuli reveal an highly significant receptive field bias in response to intact and trimmed whiskers in layer IV as well as in layers II-III neurons in only 15 h after whisker trimming. Layer IV barrel cells fail to show plasticity after 15-24 h in a standard cage environment, but produce a response bias when activity is elevated by the enriched environment. We conclude that elevated activity achieves the threshold for response modification more quickly, and this, in turn, accelerates the rate of receptive field plasticity.

Effect of enriched environment rearing on impairments in cortical excitability and plasticity after prenatal alcohol exposure.

The daily ingestion of alcohol by pregnant mammals exposes the fetal brain to varying levels of alcohol through the placental circulation. Here we focus on the lingering impact on cortical function of 6.5% alcohol administered in a liquid diet to pregnant rats throughout gestation, followed by 3 alcohol-free months before brain function was analyzed in the offspring. Both spontaneous activity of the neurons in the barrel cortex and the level of response to test stimuli applied to the whiskers remained reduced by >75% after alcohol exposure. Whisker pairing, a type of cortical plasticity induced by trimming all but two whiskers in adult rats, occurred in <1 d in controls, but required 14 d to reach significance after alcohol exposure. These long-term neuronal deficits are present in all layers of cortex and affect neurons with both fast and slow action potentials. Plasticity is first seen in the total sample of neurons at 14 d; however, by 7 d, neurons in layer II/III already show plasticity, with no change in layer IV neurons, and a reverse shift occurs toward the inactive whisker in layer V neurons. Analysis of NMDA receptor subunits shows a persistent, approximately 30-50% reduction of NR1, NR2A, and NR2B subunits at postnatal day 90 in the barrel field cortex. Exposing the prenatal alcohol-exposed rats to enriched rearing conditions significantly improves all measured cortical functions but does not restore normal values. The results predict that combinations of interventions will be necessary to completely restore cortical function after exposure of the fetal brain to alcohol.

Lesions of mature barrel field cortex interfere with sensory processing and plasticity in connected areas of the contralateral hemisphere.

Lesions of primary sensory cortex produce impairments in brain function as an outcome of the direct tissue damage. In addition, indirect lesion effects have been described that consist of functional deficits in areas sharing neural connections with the damaged area. The present study characterizes interhemispheric deficits produced as a result of unilateral lesions of the entire vibrissa representation of S-I barrel field cortex (BFC) in adult rats using single-neuron recording under urethane anesthesia. After unilateral lesions of adult BFC, responses of neurons in the contralateral homotopic BFC are severely depressed. Background (spontaneous) activity is reduced by approximately 80%, responses to test stimuli applied to the whiskers are reduced by approximately 50%, and onset of synaptic plasticity induced by trimming all but two whiskers ("whisker-pairing plasticity") is delayed over sevenfold compared with sham-lesion control animals. These deficits persist with only slight improvement for at least 4 months after lesion. Both fast-spiking and regular-spiking neuron responses are diminished contralateral to the lesion, as are cells above, below, and within the cortical barrels. Enriched environment experience increased the magnitude of responses and accelerated the rate of synaptic plasticity but did not restore response magnitude to control levels. Deficiencies in evoked responses and synaptic plasticity are primarily restricted to areas that share direct axonal connections with the lesioned cortex, because equivalently sized lesions of visual cortex produce minimal deficits in contralateral BFC function. These results indicate that interhemispheric deficits consist of remarkable and persistent decrements in sensory processing at the single-neuron level and support the idea that the deficits are somehow linked to the shared neural connections with the area of brain damage.

Experience-dependent plasticity is impaired in adult rat barrel cortex after whiskers are unused in early postnatal life.

The capacity of adult barrel cortex to show experience-dependent plasticity after early restricted neonatal sensory deprivation was analyzed in barrel field cortex neurons. Selective sensory deprivation was induced by trimming two whiskers from postnatal day 0 (P0) to P21, namely, the principal D2 whisker plus one adjacent surround whisker (D3). At maturity (P90), responses of supragranular (layer II/III) and barrel (layer IV) neurons, all located in the D2 barrel column, were analyzed for modified responses to the deprived principal whisker (D2) and the nondeprived (D1) and deprived (D3) adjacent surround whiskers. For supragranular neurons, the responses to both principal and surround whiskers were reduced at maturity, whereas the barrel neurons showed mildly elevated responses to the principal whisker but a reduced response to the deprived surround whisker. In normal adult rats, trimming all but the principal D2 whisker and an adjacent D3 whisker for 3 d (whisker pairing) produced the expected bias: elevated responses from the intact D3 compared with the cut D1 whisker in both barrel and supragranular neurons. When the neonatally deprived D2 and D3 whiskers were paired at maturity, a similar D3/D1 bias was generated in barrel neurons, but no bias occurred in supragranular neuron responses. Pairing the maintained D1 and deprived D2 whiskers produced a much greater bias toward D1 compared with the deprived D3 whisker in barrel neurons than in supragranular neurons. There were minimal effects on response latencies in layer IV under any of the experimental conditions. These findings indicate that a restricted period of sensory deprivation in early postnatal life (1) impairs intracortical relay of deprived inputs from layer IV to layer II/III in barrel cortex at maturity and (2) degrades receptive field plasticity of the supragranular layer cells but not the thalamic-recipient barrel neurons.

Molecular cloning and expression of a protein-tyrosine phosphatase showing homology with transcription factors Fos and Jun.

A cDNA clone coding for a protein-tyrosine phosphatase (PTPase) was isolated from a rat spleen cDNA library. Nucleotide sequence of the clone showed an open reading frame coding for a polypeptide of 363 amino acids. Expression of this clone in E. coli in an expression vector showed PTPase activity. The non-catalytic region of this PTPase located at the carboxy terminus shows homology with the basic domains of transcription factors Fos and Jun. Northern blot analysis showed that a 1.7 kb transcript was present in many tissues and cells, the highest level being in macrophages. This PTPase is a rat homolog of human T-cell PTPase although it shows 3 large deletions in the carboxy terminal non-catalytic region.

Postnatal changes in NMDAR1 subunit expression in the rat trigeminal pathway to barrel field cortex.

N-methyl-D-aspartate (NMDA) type glutamate receptors are constituted of one obligatory subunit (NR1), expressed as eight splice variants, combined with one or more of four NMDAR2 subunits. Polyclonal antibodies were produced to an N-terminal domain of the NR1 subunit that recognize all eight splice variants. The antibody was used to localize NR1 in the trigeminal pathway to barrel field cortex in rats. The distribution and density of NR1 changes between birth (postnatal day 0 = P-0) and P-360. The trigeminal nuclei already contain a high level of NR1 immunoreactivity on the day of birth. The ventral posterior lateral, ventral posterior medial, and posterior nucleus, medial division, thalamic nuclei show fluctuations in NR1 immunoreactivity levels, starting at birth with moderate densities in neuropil which decrease at P-7, and peak again in neuronal cell bodies as well as the neuropil at P-21. In the cortex, the density of NR1 in layer VI fluctuates with low points at P-7 and P-40. Superficial cortical layers I, II, and III reach adult levels at P-14 and remain high. NR1 levels decrease sharply in layer IV just prior to P-40 and then slowly recover over the next 3 months to stabilize at moderate levels in the adult. In addition to neuronal expression there is a transient high level of labeling in glial cells with a peak density of staining at P-21. The results emphasize that NR1 subunit expression is finely regulated in rat somatic sensory pathways for periods as long as 7-8 weeks after birth in the barrel field cortex.

Deregulation of tyrosine kinases in a rat macrophage tumor - ectopic expression of lck.

AK-5, a spontaneous rat macrophage tumor has 2-3 fold higher membrane-associated tyrosine kinase (TK) activity than normal peritoneal macrophages. lck, the lymphoid-specific TK, is expressed ectopically in AK-5. p56(lck), the protein product of lck, was detected in AK-5 by immunoprecipitation. The surface proteins CD4 and CD8 to which p56(lck) is complexed in lymphocytes are not present an the surface of AK-5. The transcripts of hck, a gene expressed predominantly in myeloid cells are markedly reduced in AK-5. The genomic organization of lck is unaltered. Although the total phosphotyrosine content of proteins is not altered significantly, there is a qualitative change in the profile of phosphotyrosine containing proteins. These results provide evidence for the ectopic expression of lck in a hematopoietic tumor and bolster the existing data implicating the participation of src family TKs in spontaneous neoplasms.

Bleach sedimentation method for increased sensitivity of sputum smear microscopy: does it work?

SETTING: A non-governmental organisation (NGO) supported tuberculosis control programme in Bangladesh with good smear microscopy. OBJECTIVE: To verify whether bleach sedimentation method increases the sensitivity of sputum smear microscopy for acid-fast bacilli (AFB), and if so, how. DESIGN: Duplicate smears from successive routine specimens, peripheral centres examining direct smears, and blind examination of bleach sediment smears at central laboratories. RESULTS: When all 3,287 sputum samples were examined in duplicate and the International Union Against Tuberculosis and Lung Disease cut-off for positivity was applied, more positives were not found by bleach sedimentation. Using the much lower American Thoracic Society (ATS) threshold, the percentage positives rose slightly from 15.5% for direct smear to 16.6% after bleach. The gain was more evident when suspect examinations only were taken into consideration, as bleach missed many positives identified by direct follow-up smear. When patients rather than individual smears were counted, more suspects were detected by bleach (10% gain on average), but with considerable variation between the centres (range 6-16%). To arrive at this gain, the ATS cut-off was used, with corrections for false results. Under routine conditions, however, this threshold is too low in view of possible transfer of AFB. CONCLUSIONS: Bleach sedimentation can increase the diagnostic yield, but only to a minor extent if all other factors have been optimised already; it is not a panacea. Precautions against false negatives as well as false positives should be taken, and the additional workload is not negligible.

Molecular cloning of a membrane bound tyrosine-specific protein kinase from rat spleen.

Tyrosine-specific protein phosphorylation is believed to play an important (though poorly understood) role in various cellular functions in many normal and malignant cells. In order to understand the function of tyrosine-specific protein kinases in normal cells, it is necessary, as an initial step, to identify genes (and proteins) for these enzymes. For this purpose cDNA libraries were constructed in plasmid vector pGEM-3Z and lambda gt11 using mRNA from rat spleen. From these cDNA libraries, cDNA clones coding for a src-related tyrosine-specific protein kinase were isolated. The largest clone (L115) was 1.94 kb in size. Various restriction fragments of this clone were subcloned in plasmid vector for sequencing. The complete nucleotide sequence of the largest clone showed an open reading frame coding for a protein of 503 amino acids. The presence of a glycine at position 2 and an arginine at position 7 indicated that this protein is likely to be acylated at glycine 2 and therefore associated with plasma membrane. This gene showed high homology to human and mouse hck and hence it is perhaps the rat homologue of hck. Moderate level of expression of this gene was observed only in the adult rat spleen and not in other tissues. These results suggest that this kinase gene is expressed in a tissue specific manner.

Physiological slowing and upregulation of inhibition in cortex are correlated with behavioral deficits in protein malnourished rats.

Protein malnutrition during early development has been correlated with cognitive and learning disabilities in children, but the neuronal deficits caused by long-term protein deficiency are not well understood. We exposed rats from gestation up to adulthood to a protein-deficient (PD) diet, to emulate chronic protein malnutrition in humans. The offspring exhibited significantly impaired performance on the 'Gap-crossing' (GC) task after reaching maturity, a behavior that has been shown to depend on normal functioning of the somatosensory cortex. The physiological state of the somatosensory cortex was examined to determine neuronal correlates of the deficits in behavior. Extracellular multi-unit recording from layer 4 (L4) neurons that receive direct thalamocortical inputs and layers 2/3 (L2/3) neurons that are dominated by intracortical connections in the whisker-barrel cortex of PD rats exhibited significantly low spontaneous activity and depressed responses to whisker stimulation. L4 neurons were more severely affected than L2/3 neurons. The response onset was significantly delayed in L4 cells. The peak response latency of L4 and L2/3 neurons was delayed significantly. In L2/3 and L4 of the barrel cortex there was a substantial increase in GAD65 (112% over controls) and much smaller increase in NMDAR1 (12-20%), suggesting enhanced inhibition in the PD cortex. These results show that chronic protein deficiency negatively affects both thalamo-cortical and cortico-cortical transmission during somatosensory information processing. The findings support the interpretation that sustained protein deficiency interferes with features of cortical sensory processing that are likely to underlie the cognitive impairments reported in humans who have suffered from prolonged protein deficiency.

Persistent sacrococcygeus ventralis muscle in an adult human pelvic wall: a variation for surgeons to note.

Occurrence of abnormal muscles in the pelvic wall is very rare. During a routine dissection of the pelvic wall, an abnormal muscle referred to as sacrococcygeus ventralis was noted in a 65-year-old South Indian cadaver. The fleshy fibers of the muscle were arising from the lateral part of the ventral surface of the sacrum at the level of S3 segment. The muscle passed downwards in front of the S4 and S5 sacral segments, halfway through its course it became tendinous and finally became inserted in the ventral surface of the coccyx. Sacrococcygeus ventralis is a muscle which is well developed in animals where it acts on their tail. In human beings, sacrococcygeus ventralis is seen only during fetal life. A rare case of its persistence in an adult pelvic wall is reported and discussed here.

Chronic suppression of activity in barrel field cortex downregulates sensory responses in contralateral barrel field cortex.

Numerous lines of evidence indicate that neural information is exchanged between the cerebral hemispheres via the corpus callosum. Unilateral ablation lesions of barrel field cortex (BFC) in adult rats induce strong suppression of background and evoked activity in the contralateral barrel cortex and significantly delay the onset of experience-dependent plasticity. The present experiments were designed to clarify the basis for these interhemispheric effects. One possibility is that degenerative events, triggered by the lesion, degrade contralateral cortical function. Another hypothesis, alone or in combination with degeneration, is that the absence of interhemispheric activity after the lesion suppresses contralateral responsiveness. The latter hypothesis was tested by placing an Alzet minipump subcutaneously and connecting it via a delivery tube to a cannula implanted over BFC. The minipump released muscimol, a GABA(A) receptor agonist at a rate of 1 mul/h, onto one barrel field cortex for 7 days. Then with the pump still in place, single cells were recorded in the contralateral BFC under urethan anesthesia. The data show a approximately 50% reduction in principal whisker responses (D2) compared with controls, with similar reductions in responses to the D1 and D3 surround whiskers. Despite these reductions, spontaneous firing is unaffected. Fast spiking units are more sensitive to muscimol application than regular spiking units in both the response magnitude and the center/surround ratio. Effects of muscimol are also layer specific. Layer II/III and layer IV neurons decrease their responses significantly, unlike layer V neurons that fail to show significant deficits. The results indicate that reduced activity in one hemisphere alters cortical excitability in the other hemisphere in a complex manner. Surprisingly, a prominent response decrement occurs in the short-latency (3-10 ms) component of principal whisker responses, suggesting that suppression may spread to neurons dominated by thalamocortical inputs after interhemispheric connections are inactivated. Bilateral neurological impairments have been described after unilateral stroke lesions in the clinical literature.

Purification and characterization of a tyrosine-specific protein kinase of Mr 60,000 and comparison with a kinase of Mr 56,000 from rat spleen.

A tyrosine-specific protein kinase of Mr 60,000 (TK-I) was purified to near homogeneity from the particulate fraction of rat spleen. The purification procedure involved sequential chromatography of the detergent-solubilized enzyme on DEAE-Sephacel and hydroxyapatite columns. Polyacrylamide-gel electrophoresis under denaturing conditions showed one major polypeptide, of Mr 60,000. Gel filtration of the enzyme on Sephacryl S-200 column showed a single peak of kinase activity, of apparent Mr 60,000. On incubation with [gamma-32P]ATP, it showed a phosphoprotein of Mr 60,000 as a result of autophosphorylation. The autophosphorylation of the kinase occurred only at tyrosine residues. Incubation of TK-I with ATP (but not with ADP) resulted in an increase in its tyrosine-specific protein kinase activity. The time course of autophosphorylation of TK-I was very similar to the time course of activation by ATP. These and other experiments suggest that autophosphorylation might be responsible for activation of TK-I observed on incubation with ATP. A second tyrosine-specific protein kinase (TK-II) was isolated from the particulate fraction of rat spleen. A highly purified preparation of TK-II on incubation with [gamma-32P]ATP gave a major phosphoprotein, of Mr 56,000. TK-II was different from TK-I in several properties: (a) substrate specificity; (b) chromatographic behaviour; (c) phosphopeptide maps; and (d) inhibition by tosyl-lysylchloromethane. Antisera raised against TK-I did not cross-react with TK-II. These results suggest that TK-I and TK-II are distinct proteins, perhaps coded by two different genes.

Experience-dependent plasticity of adult rat S1 cortex requires local NMDA receptor activation.

The effect of blocking NMDA glutamate receptors in adult rat cortex on experience-dependent synaptic plasticity of barrel cortex neurons was studied by infusing D-AP5 with an osmotic minipump over barrel cortex for 5 d of novel sensory experience. In acute pilot studies, 500 microM D-AP5 was shown to specifically suppress NMDA receptor (NMDAR)-dependent responses of single cells in cortical layers I-IV. To induce plasticity, all whiskers except D2 and D1 were cut close to the face 1 d after pump insertion. The animals were housed with 2 cage mates before recording 4 d later. This pairing of two whiskers for several days in awake animals generates highly significant biases in responses from D2 layer IV (barrel) cells to the intact D1 whisker as opposed to the cut D3 whisker. D-AP5 completely prevented the D1/D3 surround whisker bias from occurring in the D2 barrel cells (p > 0.6 for D1 > D3, Wilcoxon). Fast-spike and slow-spike barrel cells were affected equally, suggesting parity for inhibitory and excitatory cell plasticity. D-AP5 only partially suppressed the D1/D3 bias in supragranular layers (layers II-III) in the same penetrations (p < 0.042 for D1 > D3). In control animals, the inactive L-AP5 isomer allowed the bias to develop normally toward the intact surround whisker (p < 0.001 for D1 > D3) for cells in all layers. We conclude that experience-dependent synaptic plasticity of mature barrel cortex is cortically dependent and that modification of local cortical NMDARs is necessary for its expression.

Theory for normal and impaired experience-dependent plasticity in neocortex of adult rats.

We model experience-dependent plasticity in the cortical representation of whiskers (the barrel cortex) in normal adult rats, and in adult rats that were prenatally exposed to alcohol. Prenatal exposure to alcohol (PAE) caused marked deficits in experience-dependent plasticity in a cortical barrel-column. Cortical plasticity was induced by trimming all whiskers on one side of the face except two. This manipulation produces high activity from the intact whiskers that contrasts with low activity from the cut whiskers while avoiding any nerve damage. By a computational model, we show that the evolution of neuronal responses in a single barrel-column after this sensory bias is consistent with the synaptic modifications that follow the rules of the Bienenstock, Cooper, and Munro (BCM) theory. The BCM theory postulates that a neuron possesses a moving synaptic modification threshold, theta(M), that dictates whether the neuron's activity at any given instant will lead to strengthening or weakening of its input synapses. The current value of theta(M) changes proportionally to the square of the neuron's activity averaged over some recent past. In the model of alcohol impaired cortex, the effective theta(M) has been set to a level unattainable by the depressed levels of cortical activity leading to "impaired" synaptic plasticity that is consistent with experimental findings. Based on experimental and computational results, we discuss how elevated theta(M) may be related to (i) reduced levels of neurotransmitters modulating plasticity, (ii) abnormally low expression of N-methyl-d-aspartate receptors (NMDARs), and (iii) the membrane translocation of Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) in adult rat cortex subjected to prenatal alcohol exposure.