The development and activity-dependent expression of aggrecan in the cat visual cortex.

The Cat-301 monoclonal antibody identifies aggrecan, a chondroitin sulfate proteoglycan in the cat visual cortex and dorsal lateral geniculate nucleus (dLGN). During development, aggrecan expression increases in the dLGN with a time course that matches the decline in plasticity. Moreover, examination of tissue from selectively visually deprived cats shows that expression is activity dependent, suggesting a role for aggrecan in the termination of the sensitive period. Here, we demonstrate for the first time that the onset of aggrecan expression in area 17 also correlates with the decline in experience-dependent plasticity in visual cortex and that this expression is experience dependent. Dark rearing until 15 weeks of age dramatically reduced the density of aggrecan-positive neurons in the extragranular layers, but not in layer IV. This effect was reversible as dark-reared animals that were subsequently exposed to light showed normal numbers of Cat-301-positive cells. The reduction in aggrecan following certain early deprivation regimens is the first biochemical correlate of the functional changes to the γ-aminobutyric acidergic system that have been reported following early deprivation in cats.

PLC-beta1, activated via mGluRs, mediates activity-dependent differentiation in cerebral cortex.

During development of the cerebral cortex, the invasion of thalamic axons and subsequent differentiation of cortical neurons are tightly coordinated. Here we provide evidence that glutamate neurotransmission triggers a critical signaling mechanism involving the activation of phospholipase C-beta1 (PLC-beta1) by metabotropic glutamate receptors (mGluRs). Homozygous null mutation of either PLC-beta1 or mGluR5 dramatically disrupts the cytoarchitectural differentiation of 'barrels' in the mouse somatosensory cortex, despite segregation in the pattern of thalamic innervation. Furthermore, group 1 mGluR-stimulated phosphoinositide hydrolysis is dramatically reduced in PLC-beta1-/- mice during barrel development. Our data indicate that PLC-beta1 activation via mGluR5 is critical for the coordinated development of the neocortex, and that presynaptic and postsynaptic components of cortical differentiation can be genetically dissociated.

Cortical plasticity: is it time for a change?

Classical studies of plasticity in the visual cortex have been interpreted in terms of heterosynaptic competition between inputs. But an alternative type of 'homosynaptic' plasticity can explain many recent observations and has recently received experimental support. Perhaps both types of plasticity are important.

Plasticity: downstream of glutamate.

Glutamate neurotransmission is an essential component of many forms of neuronal plasticity, however, the intracellular mechanisms that mediate plasticity are only beginning to be elucidated. The emerging image of the NMDA receptor complex reminds us that the similarity between mechanisms of plasticity in various model systems is greater than their apparent differences. For example, the cAMP-dependent protein kinase A signalling pathway is crucial for plasticity in a variety of neuronal systems and across a wide variety of species.

Neural activity: sculptor of 'barrels' in the neocortex.

A major portion of the primary somatosensory cortex of rodents is characterized by the discrete and patterned distribution of thalamocortical axons and layer IV granule cells ('barrels'), which correspond to the spatial distribution of whiskers and sinus hairs on the snout. In recent years several mutant mouse models began unveiling the cellular and molecular mechanisms by which these patterns emerge presynaptically and are reflected postsynaptically. Neural activity plays a crucial role in conferring presynaptic patterns to postsynaptic cells via neurotransmitter receptor-mediated intracellular signals. Here we review recent evidence that is finally opening the doors to understanding the cellular and molecular mechanisms of pattern formation in the neocortex.

Effects of early periods of monocular deprivation and reverse lid suture on the development of Cat-301 immunoreactivity in the dorsal lateral geniculate nucleus (dLGN) of the cat.

During certain sensitive periods early in postnatal life, the anatomical and physiological development of the central visual pathways of cats and monkeys can be affected by the nature of an animal's early visual experience. In the last few years, studies have been started on some of the molecular and biochemical events that underlie the many functional changes induced by early selected visual deprivation in the visual cortex of kittens. In this respect, the monoclonal antibody Cat-301 provides a potentially powerful tool, because it recognizes in the cat dorsal lateral geniculate nucleus (dLGN) a proteoglycan associated with the surface of a particular class of cells, namely Y cells. In the dLGN, the Cat-301 proteoglycan appears late in postnatal development, and it expression has been shown to be experience dependent in both the dLGN and visual cortex (M. Sur, D. Frost, and S. Hockfield, 1988, J. Neurosci. 8:874-882; A. Guimaraes, S. Zaremba, and S. Hockfield, 1990, J. Neurosci. 10:3014-3024). We have explored further the experience-dependent nature of Cat-301 expression in the dLGN with a view to establishing a biochemical correlate of the many functional changes induced by early monocular deprivation and its reversal in the kitten visual system. In addition to demonstrating differences in Cat-301 expression between deprived and nondeprived laminae of the dLGNs of kittens monocularly deprived to only 4 or 5 weeks of age, the magnitude of the laminar difference was found to increase as the period of deprivation was extended. Moreover, monocularly deprived kittens that subsequently received long periods of reverse lid suture exhibited a reversal of the pattern of immunoreactivity, so that the greatest immunoreactivity occurred in laminae innervated by the initially deprived eye. However, possibly the most surprising and important finding was the extremely low levels of immunoreactivity observed in both A laminae of monocularly deprived animals that had received relatively short periods of reverse lid suture. These data suggest that Y cell development can be drastically altered depending on the time of initiation of the period of reverse lid suture and its duration.

Phospholipase C-beta1 expression correlates with neuronal differentiation and synaptic plasticity in rat somatosensory cortex.

Receptor-mediated signal transduction is thought to play an important role in neuronal differentiation and the modification of synaptic connections during brain development. The intracellular signalling molecule phospholipase C-beta1 (PLC-beta1), which is activated via specific neurotransmitter receptors, has recently been implicated in activity-dependent plasticity in the cat visual cortex. PLC-beta1 has been shown to be concentrated in an intermediate compartment-like organelle, the botrysome, which is present in 5-week-old, but not adult, cat cortical neurons. We have characterized the spatial and temporal regulation of PLC-beta1 expression in the developing rat cerebral cortex. PLC-beta1-positive botrysome-like organelles are observed during early postnatal cortical development, but not at postnatal day 14 or later stages. In the postnatal somatosensory cortex, there is also striking spatial variation in diffuse neuropilar immunoreactivity of layer IV and above, in a pattern corresponding to the thalamocortical recipient zones known as barrels. This expression pattern is specific to the developing barrel field and is most distinct at postnatal days 4-7, when cellular components of barrels are capable of activity-dependent modification. During later stages of cortical maturation, stained botrysomes disappear, expression of PLC-beta1 is down-regulated and only diffuse immunoreactivity remains in dendritic processes. Our results are consistent with a role for PLC-beta1 in activity-dependent, receptor-mediated neuronal plasticity during development of the somatosensory cortex.

SynGAP isoforms exert opposing effects on synaptic strength.

Alternative promoter usage and alternative splicing enable diversification of the transcriptome. Here we demonstrate that the function of Synaptic GTPase-Activating Protein (SynGAP), a key synaptic protein, is determined by the combination of its amino-terminal sequence with its carboxy-terminal sequence. 5' rapid amplification of cDNA ends and primer extension show that different N-terminal protein sequences arise through alternative promoter usage that are regulated by synaptic activity and postnatal age. Heterogeneity in C-terminal protein sequence arises through alternative splicing. Overexpression of SynGAP α1 versus α2 C-termini-containing proteins in hippocampal neurons has opposing effects on synaptic strength, decreasing and increasing miniature excitatory synaptic currents amplitude/frequency, respectively. The magnitude of this C-terminal-dependent effect is modulated by the N-terminal peptide sequence. This is the first demonstration that activity-dependent alternative promoter usage can change the function of a synaptic protein at excitatory synapses. Furthermore, the direction and degree of synaptic modulation exerted by different protein isoforms from a single gene locus is dependent on the combination of differential promoter usage and alternative splicing.

Monoclonal antibody Cat-301 selectively identifies a subset of nuclei in the cat's somatosensory thalamus.

Recently it has been demonstrated that the monoclonal antibody Cat-301 is capable of identifying functionally related neurons in the mammalian visual thalamus. We have examined the possibility that this antibody might display a similar capacity in nonvisual thalamic areas. We demonstrate that in the cat's somatosensory thalamus the distribution of Cat-301-positive cells and neuropil is restricted to a subset of nuclei. These include the ventroposterior medial, ventroposterior lateral, and ventroposterior inferior nuclei. Staining with Cat-301 provides a clear visualisation of the entire somatotopic map within these nuclei. The somatosensory sector of the thalamic reticular nucleus and the perireticular nucleus, which may have a somatosensory sector, are also Cat-301-positive. In contrast, cells that do not express the Cat-301 antigen are located in the ventroposterior oralis nucleus, the ventroposterior shell region, the medial and lateral divisions of the posterior nuclear group, and the inner small cell region adjacent to the thalamic reticular nucleus. In comparison with previous physiological studies, cells that express the Cat-301 antigen most likely represent subpopulations in only a few of the somatic submodality-specific groups. These include cells in the small-field and Pacinian cutaneous-responsive groups, excluding cells in the wide-field cutaneous-, muscle-, joint-, and noxious-responsive groups. Taken together these findings indicate that monoclonal antibody Cat-301 is capable of selectively identifying neurons with distinct functional properties in the mammalian somatosensory thalamus.

Interocular suppression in the visual cortex of strabismic cats.

Strabismic humans usually experience powerful suppression of vision in the nonfixating eye. In an attempt to demonstrate physiological correlates of such suppression, we recorded from the primary visual cortex of cats with surgically induced squint and studied the responses of neurons to drifting gratings of different orientation, spatial frequency, and contrast in the two eyes. Only 1 of 50 apparently monocular cells showed any evidence of remaining, subliminal excitatory input from the "silent" eye when the two eyes were stimulated with gratings of similar orientation, and even among the small proportion of cells that remained binocularly driven, very few exhibited facilitation when stimulated binocularly. The majority of cells from both exotropes and esotropes, even those that could be independently driven through either eye, displayed nonspecific interocular suppression: stimulation of the nondominant eye with a drifting grating of any orientation depressed the response to an optimal grating being presented to the dominant eye. This phenomenon exhibited a gross nonlinearity in that it was dependent on the temporal sequence of stimulus presentation: stimulation of the nondominant eye caused significant suppression only if the neuron was already responding to an appropriate stimulus in the dominant eye, but not when onset of stimulation in the two eyes was simultaneous. Interocular suppression was always independent of the relative spatial phase of the two grating stimuli, and usually broadly tuned for the spatial frequency of the suppressive stimulus. Suppression may depend on inhibitory interaction between neighboring ocular dominance columns, combined with the loss of conventional disparity-selective binocular interactions for matched stimuli in the two eyes. The similarity of interocular suppression in strabismic cats and that caused by orthogonal gratings in the two eyes in normal cats (Sengpiel and Blakemore, 1994; Sengpiel et al., 1994) suggests that strabismic suppression and binocular rivalry depend on similar neural mechanisms.

Initial recovery of vision after early monocular deprivation in kittens is faster when both eyes are open.

A comparison was made of the speed of visual recovery in the deprived eye of kittens after a 6-day period of monocular deprivation imposed at 5-9 weeks of age in two postdeprivation conditions. In one condition, binocular recovery (BR), both eyes were open, whereas in the other condition, reverse lid-suture (RLS), the formerly nondeprived eye was closed to force the animal to use the originally deprived eye. In littermate pairs, BR kittens began to recover form vision 12 to 30 h before those subjected to RLS. The vision of the deprived eye of the BR animals remained superior to that of their RLS littermates for 4-8 days. Although this finding is difficult to reconcile with competitive mechanisms of synaptic plasticity, it supports a prediction of an alternative model of synaptic plasticity [Bienenstock, E. L., Cooper, L. N. & Munro, P. W. (1982) J. Neurosci. 2, 32-48] for slower initial recovery with RLS because of the time required to reset the modification threshold.

Phospholipase C-beta1 is present in the botrysome, an intermediate compartment-like organelle, and Is regulated by visual experience in cat visual cortex.

Monoclonal antibody Cat-307 identifies a 165 kDa neuronal protein expressed in the cat visual cortex during the period of sensitivity to alterations in visual experience (). Dark-rearing, which prolongs the sensitive period, also prolongs the expression of the Cat-307 protein. The Cat-307 protein localizes to an organelle, here called the botrysome (from the Greek botrys, cluster of grapes), that is located between the endoplasmic reticulum (ER) and Golgi apparatus. The botrysome is composed of small ring-shaped profiles with electron-dense coats. The size and morphology of the rings and their coats are similar to those described for ER to Golgi transport vesicles. Biochemically, the Cat-307 protein cofractionates with microsomes and partitions with subunits of the coatomer proteins that coat ER-to-Golgi transport vesicles. Partial amino acid sequencing reveals that the Cat-307 protein is phospholipase C-beta1, the G-protein-dependent phosphodiesterase that hydrolyses phosphatidylinositol 4,5 biphosphate into inositol 1,4,5 triphosphate and diacylglycerol after the stimulation of a variety of neurotransmitter receptors at the cell surface. These results suggest a role for phospholipase C-beta1 and the botrysome in developmental plasticity and provide a possible link between receptor activation at the cell surface and protein transport during neuronal development.

Functional architecture of area 17 in normal and monocularly deprived marmosets (Callithrix jacchus).

The organization of the primary visual cortex (VI) of the common marmoset (Callithrix jacchus) was studied both physiologically and by means of transneuronal labelling of geniculocortical afferents. We addressed the question whether monocular deprivation (MD) could stabilize segregation into ocular dominance (OD) columns, which are not seen in normal adult marmosets but are present in juvenile animals (Spatz, 1979, 1989). Properties of neurons in normal marmosets closely resembled those of other New-World and Old-World monkeys and orderly tangential progressions of preferred orientation were observed. However, in contrast to species that display well-defined OD columns, neurons of layer 4 in V1 of normal adult marmosets received balanced inputs from the two eyes. Early MD (even though followed by prolonged binocular experience into adulthood) resulted in a reduction of cell size in laminae of the lateral geniculate nucleus with input from the deprived eye and a dramatic overall shift in ocular dominance towards the nondeprived eye in the cortex. However, isolated clusters of cells dominated by the deprived eye were found in both layers 4 and 6. Injection of lectin-conjugated horseradish peroxidase (WGA-HRP) into the deprived eye revealed elongated patches of terminal label, about 350 microns wide, in flat-mounted sections through layer 4. Afferent segregation was sharper and more regular in the region of V1 representing parafoveal visual space than in that representing the fovea. Our findings support the notion that all Old-World and New-World monkeys possess the capacity for segregation of geniculocortical afferents into OD columns.