A biochemical correlate of the critical period for synaptic modification in the visual cortex.

Stimulation of phosphoinositide hydrolysis by excitatory amino acids was studied in synaptoneurosomes of kitten striate cortex at several postnatal ages. Ibotenate and glutamate stimulated phosphoinositide turnover during the second and third postnatal months; N-methyl-D-aspartate and DL-alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) were without effect. The developmental profile of ibotenate-stimulated phosphoinositide turnover parallels the postnatal changes in cortical susceptibility to visual deprivation. The transient increase in ibotenate-stimulated phosphoinositide turnover does not occur in visual cortex of kittens reared in complete darkness.

Blockade of "NMDA" receptors disrupts experience-dependent plasticity of kitten striate cortex.

Intracortical infusion of the "N-methyl-D-aspartate" (NMDA) receptor blocker D,L-2-amino-5-phosphonovaleric acid (APV) renders kitten striate cortex resistant to the effects of monocular deprivation. In addition, 1 week of continuous APV treatment (50 nanomoles per hour) produces a striking loss of orientation selectivity in area 17. These data support the hypothesis that crucial variables for the expression of activity-dependent synaptic modifications are a critical level of postsynaptic activation and calcium entry through ion channels linked to NMDA receptors.

A physiological basis for a theory of synapse modification.

The functional organization of the cerebral cortex is modified dramatically by sensory experience during early postnatal life. The basis for these modifications is a type of synaptic plasticity that may also contribute to some forms of adult learning. The question of how synapses modify according to experience has been approached by determining theoretically what is required of a modification mechanism to account for the available experimental data in the developing visual cortex. The resulting theory states precisely how certain variables might influence synaptic modifications. This insight has led to the development of a biologically plausible molecular model for synapse modification in the cerebral cortex.

Role for rapid dendritic protein synthesis in hippocampal mGluR-dependent long-term depression.

A hippocampal pyramidal neuron receives more than 10(4) excitatory glutamatergic synapses. Many of these synapses contain the molecular machinery for messenger RNA translation, suggesting that the protein complement (and thus function) of each synapse can be regulated on the basis of activity. Here, local postsynaptic protein synthesis, triggered by synaptic activation of metabotropic glutamate receptors, was found to modify synaptic transmission within minutes.

Common forms of synaptic plasticity in the hippocampus and neocortex in vitro.

Activity-dependent synaptic plasticity in the superficial layers of juvenile cat and adult rat visual neocortex was compared with that in adult rat hippocampal field CA1. Stimulation of neocortical layer IV reliably induced synaptic long-term potentiation (LTP) and long-term depression (LTD) in layer III with precisely the same types of stimulation protocols that were effective in CA1. Neocortical LTP and LTD were specific to the conditioned pathway and, as in the hippocampus, were dependent on activation of N-methyl-D-aspartate receptors. These results provide strong support for the view that common principles may govern experience-dependent synaptic plasticity in CA1 and throughout the superficial layers of the mammalian neocortex.

Bidirectional, activity-dependent regulation of glutamate receptors in the adult hippocampus in vivo.

Experience-dependent regulation of synaptic strength has been suggested as a physiological mechanism by which memory storage occurs in the brain. Although modifications in postsynaptic glutamate receptor levels have long been hypothesized to be a molecular basis for long-lasting regulation of synaptic strength, direct evidence obtained in the intact brain has been lacking. Here we show that in the adult brain in vivo, synaptic glutamate receptor trafficking is bidirectionally, and reversibly, modified by NMDA receptor-dependent synaptic plasticity and that changes in glutamate receptor protein levels accurately predict changes in synaptic strength. These findings support the idea that memories can be encoded by the precise experience-dependent assignment of glutamate receptors to synapses in the brain.

Visual experience and deprivation bidirectionally modify the composition and function of NMDA receptors in visual cortex.

The receptive fields of visual cortical neurons are bidirectionally modified by sensory deprivation and experience, but the synaptic basis for these changes is unknown. Here we demonstrate bidirectional, experience-dependent regulation of the composition and function of synaptic NMDA receptors (NMDARs) in visual cortex layer 2/3 pyramidal cells of young rats. Visual experience decreases the proportion of NR2B-only receptors, shortens the duration of NMDAR-mediated synaptic currents, and reduces summation of synaptic NMDAR currents during bursts of high-frequency stimulation. Visual deprivation exerts an opposite effect. Although the effects of experience and deprivation are reversible, the rates of synaptic modification vary. Experience can induce a detectable change in synaptic transmission within hours, while deprivation-induced changes take days. We suggest that experience-dependent changes in NMDAR composition and function regulate the development of receptive field organization in visual cortex.

NMDA induces long-term synaptic depression and dephosphorylation of the GluR1 subunit of AMPA receptors in hippocampus.

Brief bath application of N-methyl-D-aspartate (NMDA) to hippocampal slices produces long-term synaptic depression (LTD) in CA1 that is (1) sensitive to postnatal age, (2) saturable, (3) induced postsynaptically, (4) reversible, and (5) not associated with a change in paired pulse facilitation. Chemically induced LTD (Chem-LTD) and homosynaptic LTD are mutually occluding, suggesting a common expression mechanism. Using phosphorylation site-specific antibodies, we found that induction of chem-LTD produces a persistent dephosphorylation of the GluR1 subunit of AMPA receptors at serine 845, a cAMP-dependent protein kinase (PKA) substrate, but not at serine 831, a substrate of protein kinase C (PKC) and calcium/calmodulin-dependent protein kinase II (CaMKII). These results suggest that dephosphorylation of AMPA receptors is an expression mechanism for LTD and indicate an unexpected role of PKA in the postsynaptic modulation of excitatory synaptic transmission.

Involvement of a postsynaptic protein kinase A substrate in the expression of homosynaptic long-term depression.

Hippocampal N-methyl-D-aspartate (NMDA) receptor-dependent long-term synaptic depression (LTD) is associated with a persistent dephosphorylation of the GluR1 subunit of AMPA receptors at a site (Ser-845) phosphorylated by cAMP-dependent protein kinase (PKA). In the present study, we show that dephosphorylation of a postsynaptic PKA substrate may be crucial for LTD expression. PKA activators inhibited both AMPA receptor dephosphorylation and LTD. Injection of a cAMP analog into postsynaptic neurons prevented LTD induction and reversed previously established homosynaptic LTD without affecting baseline synaptic transmission. Moreover, infusing a PKA inhibitor into postsynaptic cells produced synaptic depression that occluded homosynaptic LTD. These findings suggest that dephosphorylation of a PKA site on AMPA receptors may be one mechanism for NMDA receptor-dependent homosynaptic LTD expression.

Rapid, experience-dependent expression of synaptic NMDA receptors in visual cortex in vivo.

Sensory experience is crucial in the refinement of synaptic connections in the brain during development. It has been suggested that some forms of experience-dependent synaptic plasticity in vivo are associated with changes in the complement of postsynaptic glutamate receptors, although direct evidence has been lacking. Here we show that visual experience triggers the rapid synaptic insertion of new NMDA receptors in visual cortex. The new receptors have a higher proportion of NR2A subunits and, as a consequence, different functional properties. This effect of experience requires NMDA receptor activation and protein synthesis. Thus, rapid regulation of postsynaptic glutamate receptors is one mechanism for developmental plasticity in the brain. Changes in NMDA receptor expression provide a mechanism by which brief sensory experience can regulate the properties of NMDA receptor-dependent plasticity in visual cortex.

Internalization of ionotropic glutamate receptors in response to mGluR activation.

Activation of group 1 metabotropic glutamate receptors (mGluRs) stimulates dendritic protein synthesis and long-term synaptic depression (LTD), but it remains unclear how these effects are related. Here we provide evidence that a consequence of mGluR activation in the hippocampus is the rapid loss of both AMPA and NMDA receptors from synapses. Like mGluR-LTD, the stable expression of this change requires protein synthesis. These data suggest that expression of mGluR-LTD is at least partly postsynaptic, and that a functional consequence of dendritic protein synthesis is the regulation of glutamate receptor trafficking.

Metaplasticity: the plasticity of synaptic plasticity.

In this paper, we review experimental evidence for a novel form of persistent synaptic plasticity we call metaplasticity. Metaplasticity is induced by synaptic or cellular activity, but it is not necessarily expressed as a change in the efficacy of normal synaptic transmission. Instead, it is manifest as a change in the ability to induce subsequent synaptic plasticity, such as long-term potentiation or depression. Thus, metaplasticity is a higher-order form of synaptic plasticity. Metaplasticity might involve alterations in NMDA-receptor function in some cases, but there are many other candidate mechanisms. The induction of metaplasticity complicates the interpretation of many commonly studied aspects of synaptic plasticity, such as saturation and biochemical correlates.

Neocortical long-term potentiation.

LTP is a form of activity-dependent synaptic plasticity that has been investigated mainly in the hippocampus. It is considered likely that similar mechanisms may also account for aspects of naturally occurring plasticity in the neocortex. Consequently, an increasing number of studies have been devoted to the investigation of neocortical LTP. Recent results suggest that at least two forms of LTP coexist in layer III of the neocortex. One depends on NMDA-receptor activation and resembles the LTP observed in hippocampal field CA1. A second form is independent of NMDA receptors and requires activation of voltage-sensitive Ca2+ channels.

Synaptic plasticity: LTP and LTD.

Long-term potentiation (LTP) is a synaptic enhancement that follows brief, high-frequency electrical stimulation in the hippocampus and neocortex. Recent evidence suggests that induction of LTP may require, in addition to postsynaptic Ca2+ entry, activation of metabotropic glutamate receptors and the generation of diffusible intercellular messengers. A new form of synaptic plasticity, homosynaptic long-term depression (LTD) has also recently been documented, which, like LTP, requires Ca2+ entry through the NMDA receptor. Current work suggests that this LTD is a reversal of LTP, and vice versa, and that the mechanisms of LTP and LTD may converge at the level of specific phosphoproteins.

Long-term potentiation of thalamocortical transmission in the adult visual cortex in vivo.

It has been suggested that NMDA receptor-dependent synaptic strengthening, like that observed after long-term potentiation (LTP), is a mechanism by which experience modifies responses in the neocortex. We report here that patterned (theta burst) stimulation of the dorsal lateral geniculate nucleus reliably induces LTP of field potentials (FPs) evoked in primary visual cortex (Oc1) of adult rats in vivo. The response enhancement is saturable, long-lasting, and dependent on NMDA receptor activation. To determine the laminar locus of these changes, current source density (CSD) analysis was performed on FP profiles obtained before and after LTP induction. LTP was accompanied by an enhancement of synaptic current sinks located in thalamorecipient (layer IV and deep layer III) and supragranular (layers II/III) cell layers. We also examined immunocytochemical labeling for the immediate early gene zif-268 1 hr after induction of LTP. In concert with the laminar changes observed in CSD analyses, we observed a significant increase in the number of zif-268-immunopositive neurons in layers II-IV that occurred over a wide extent of Oc1. Last, we investigated the functional consequences of LTP induction by monitoring changes in visually evoked potentials. After LTP, we observed that the cortical response to a full-field flash was significantly enhanced and that responses to grating stimuli were increased across a range of spatial frequencies. These findings are consistent with growing evidence that primary sensory cortex remains plastic into adulthood, and they show that the mechanisms of LTP can contribute to this plasticity.

Developmental inhibitory gate controls the relay of activity to the superficial layers of the visual cortex.

A developmental reduction in the radial transmission of synaptic activity has been proposed to underlie the end of the critical period for experience-dependent modification in layers II/III of the visual cortex. Using paired-pulse stimulation, we investigated in visual cortical slices how the propagation of synaptic activity to the superficial layers changes during development and how this process is affected by sensory experience. The results can be summarized as follows. (1) Layers II/III responses to repetitive stimulation of the white matter become increasingly depressed between the third and sixth week of postnatal development, a time course that parallels the end of the critical period. (2) Paired-pulse depression is reduced after dark rearing and also by blocking inhibitory synaptic transmission. (3) Paired-pulse depression and its regulation by age and sensory experience is more pronounced when stimulation is applied to the white matter than when applied to layer IV. Together, these results are consistent with the idea that the maturation of intracortical inhibition reduces the capability of the cortex to relay incoming high-frequency patterns of activity to the supragranular layers.

A role for the cytoplasmic polyadenylation element in NMDA receptor-regulated mRNA translation in neurons.

The ability of neurons to modify synaptic connections based on activity is essential for information processing and storage in the brain. The induction of long-lasting changes in synaptic strength requires new protein synthesis and is often mediated by NMDA-type glutamate receptors (NMDARs). We used a dark-rearing paradigm to examine mRNA translational regulation in the visual cortex after visual experience-induced synaptic plasticity. In this model system, we demonstrate that visual experience induces the translation of mRNA encoding the alpha-subunit of calcium/calmodulin-dependent kinase II in the visual cortex. Furthermore, this increase in translation is NMDAR dependent. One potential source for newly synthesized proteins is the translational activation of dormant cytoplasmic mRNAs. To examine this possibility, we developed a culture-based assay system to study translational regulation in neurons. Cultured hippocampal neurons were transfected with constructs encoding green fluorescent protein (GFP). At 6 hr after transfection, approximately 35% of the transfected neurons (as determined by in situ hybridization) expressed detectable GFP protein. Glutamate stimulation of the cultures at this time induced an increase in the number of neurons expressing GFP protein that was NMDAR dependent. Importantly, the glutamate-induced increase was only detected when the 3'-untranslated region of the GFP constructs contained intact cytoplasmic polyadenylation elements (CPEs). Together, these findings define a molecular mechanism for activity-dependent synaptic plasticity that is mediated by the NMDA receptor and requires the CPE-dependent translation of an identified mRNA.

Therapeutic implications of the mGluR theory of fragile X mental retardation.

Evidence is reviewed that the consequences of group 1 metabotropic glutamate receptor (Gp1 mGluR) activation are exaggerated in the absence of the fragile X mental retardation protein, likely reflecting altered dendritic protein synthesis. Abnormal mGluR signaling could be responsible for remarkably diverse psychiatric and neurological symptoms in fragile X syndrome, including delayed cognitive development, seizures, anxiety, movement disorders and obesity.

Cholinergic manipulation alters stimulus-evoked metabolic activity in cat somatosensory cortex.

The role of acetylcholine (ACh) in cerebral cortical activity has recently been reevaluated. It now seems clear that this neurotransmitter increases the magnitude of cortical responses. Although substantial information has been gathered regarding the role of ACh in sensory information processing, little is known about the participation of ACh in the organization of maps in the cerebral cortex. To address this issue, we used 2 methods to manipulate the supply of ACh in the somatosensory cortex of cats: 1) unilateral neurotoxic lesions of the basal forebrain and 2) unilateral topical applications of the cholinergic antagonist, atropine. For each experimental condition, the animal received an injection of 2-deoxyglucose (2DG) while identical somatic stimuli were delivered to the right and left forepaws. In the somatosensory cortex, the 2DG uptake most often occurred in the form of patches that extended from layer II to IV. When the patches were reconstructed into 2-dimensional maps of activity throughout the somatosensory cortex, they formed strips that ran in the rostrocaudal direction. The reconstructed maps revealed that the 2DG patterns in ACh-depleted and the normal cortex were similar in their overall topographic distribution. Depletion or antagonism of ACh, however, caused the stimulus-evoked metabolic label to be reduced in dimension and density. Measurements of background activity levels were obtained by using 1) cytochrome oxidase histochemistry or 2) metabolic activity values in regions of somatosensory cortex that were not specifically stimulated. This analysis indicated that background values in the ACh-depleted hemispheres were not different from those in the normal hemispheres. The absence of ACh therefore appears to reduce the cortical response to stimulation, while background activity values do not change. These observations indicate that ACh plays a significant role in the processing of sensory information and the organization of somatosensory cortical maps.

Postnatal changes in the distribution of acetylcholinesterase in kitten striate cortex.

We have traced the postnatal development of axons and cells in kitten striate cortex that contain acetylcholinesterase (AChE) by using a modification of Koelle's histochemical method. The maturation of AChE-positive axons was not found to be fully complete until at least 3 months of age, and was characterized by several distinct developmental trends. AChE-positive fibers in layers IVc-VI proliferate rapidly after birth until, by 4 weeks postnatal, they appear to exceed the adult density. They remain at this level as late as 8 weeks and then decrease to the adult density by 13 weeks. In contrast, the AChE-positive fibers in layer I do not show a substantial increase in density until 6 weeks of age and the adult level is not achieved before 3 months postnatal. Finally, the density of AChE-positive fibers in layers II and III appears to increase gradually from birth until the mature pattern is reached at about 6 weeks. AChE could also be localized histochemically to cell bodies whose position and appearance depended on postnatal age. Stained cells first appeared in the white matter subjacent to layer VI shortly after birth. By 2 weeks of age, most cells in layer VI were also AChE positive. The staining of these cells gradually disappears over the next 2 months until, at 3 months of age, there are no AChE-positive cells in cat striate cortex. However, a subpopulation of stained neurons appears in layer V by 1 year of age that persists throughout adulthood. The possible contributions of acetylcholine and AChE to the postnatal development of kitten striate cortex are discussed.