Chronic γ-secretase inhibition reduces amyloid plaque-associated instability of pre- and postsynaptic structures.

The loss of synapses is a strong histological correlate of the cognitive decline in Alzheimer's disease (AD). Amyloid ß-peptide (Aß), a cleavage product of the amyloid precursor protein (APP), exerts detrimental effects on synapses, a process thought to be causally related to the cognitive deficits in AD. Here, we used in vivo two-photon microscopy to characterize the dynamics of axonal boutons and dendritic spines in APP/Presenilin 1 (APP(swe)/PS1(L166P))-green fluorescent protein (GFP) transgenic mice. Time-lapse imaging over 4 weeks revealed a pronounced, concerted instability of pre- and postsynaptic structures within the vicinity of amyloid plaques. Treatment with a novel sulfonamide-type γ-secretase inhibitor (GSI) attenuated the formation and growth of new plaques and, most importantly, led to a normalization of the enhanced dynamics of synaptic structures close to plaques. GSI treatment did neither affect spines and boutons distant from plaques in amyloid precursor protein/presenilin 1-GFP (APPPS1-GFP) nor those in GFP-control mice, suggesting no obvious neuropathological side effects of the drug.

[Comparison of MultiHance and Gadovist for cerebral MR perfusion imaging in healthy volunteers]. / Vergleich von MultiHance((R)) und Gadovist((R)) zur zerebralen MR-Perfusionsmessung bei gesunden Probanden.

To evaluate the weakly protein interacting MR contrast agent MultiHance((R)) and the one-molar agent Gadovist((R)) for cerebral perfusion MR imaging, a randomized intraindividual study was conducted in 12 healthy male volunteers. Perfusion-MRI was performed with single and double dose of each contrast agent on a 1.5T MR system using a gradient-echo EPI sequence. The imaging parameters, slice positioning and contrast media application were standardized. For the quantitative assessment rCBV and rCBF measurements of gray and white matter were performed. Additionally, the percentage of signal drop and the full width half maximum (FWHM) of ROI signal time curves were quantified. In a qualitative analysis the image quality of the rCBV and rCBF maps were assessed. Single dosage of the used new contrast agents was sufficient to achieve high quality perfusion maps. The susceptibility effect, described by percentage of signal loss (Gadovist((R)): 29.4% vs. MultiHance((R)): 28.3%) and the FWHM (Gadovist((R)): 6.4 s vs. Multihance((R)): 7.0 s) were not different between the agents for single dose. The one molar MR contrast agent Gadovist((R)) has no advantages over MultiHance((R)), a MR contrast agent with a higher relaxivity in perfusion MRI. Both agents allow the calculation of high quality perfusion maps at a dosage of 0.1 mmol/kg bw with physiologic absolute values for regional CBV and CBF. The susceptibility effect is comparable for both agents and stronger than with conventional MR contrast media.

Pairing-induced changes of orientation maps in cat visual cortex.

We have studied the precise temporal requirements for plasticity of orientation preference maps in kitten visual cortex. Pairing a brief visual stimulus with electrical stimulation in the cortex, we found that the relative timing determines the direction of plasticity: a shift in orientation preference toward the paired orientation occurs if the cortex is activated first visually and then electrically; the cortical response to the paired orientation is diminished if the sequence of visual and electrical activation is reversed. We furthermore show that pinwheel centers are less affected by the pairing than the pinwheel surround. Thus, plasticity is not uniformly distributed across the cortex, and, most importantly, the same spike time-dependent learning rules that have been found in single-cell in vitro studies are also valid on the level of cortical maps.

An analysis of orientation and ocular dominance patterns in the visual cortex of cats and ferrets.

We report an analysis of orientation and ocular dominance maps that were recorded optically from area 17 of cats and ferrets. Similar to a recent study performed in primates (Obermayer & Blasdel, 1997), we find that 80% (for cats and ferrets) of orientation singularities that are nearest neighbors have opposite sign and that the spatial distribution of singularities deviates from a random distribution of points, because the average distances between nearest neighbors are significantly larger than expected for a random distribution. Orientation maps of normally raised cats and ferrets show approximately the same typical wavelength; however, the density of singularities is higher in ferrets than in cats. Also, we find the well-known overrepresentation of cardinal versus oblique orientations in young ferrets (Chapman & Bonhoeffer, 1998; Coppola, White, Fitzpatrick, & Purves, 1998) but only a weak, not quite significant overrepresentation of cardinal orientations in cats, as has been reported previously (Bonhoeffer & Grinvald, 1993). Orientation and ocular dominance slabs in cats exhibit a tendency of being orthogonal to each other (Hubener, Shoham, Grinvald, & Bonhoeffer, 1997), albeit less pronounced, as has been reported for primates (Obermayer & Blasdel, 1993). In chronic recordings from single animals, a decrease of the singularity density and an increase of the ocular dominance wavelength with age but no change of the orientation wavelengths were found. Orientation maps are compared with two pattern models for orientation preference maps: bandpass-filtered white noise and the field analogy model. Bandpass-filtered white noise predicts sign correlations between orientation singularities, but the correlations are significantly stronger (87% opposite sign pairs) than what we have found in the data. Also, bandpass-filtered noise predicts a deviation of the spatial distribution of singularities from a random dot pattern. The field analogy model can account for the structure of certain local patches but not for the whole orientation map. Differences between the predictions of the field analogy model and experimental data are smaller than what has been reported for primates (Obermayer & Blasdel, 1997), which can be explained by the smaller size of the imaged areas in cats and ferrets.

Visual cortex maps are optimized for uniform coverage.

Cat visual cortex contains a topographic map of visual space, plus superimposed, spatially periodic maps of ocular dominance, spatial frequency and orientation. It is hypothesized that the layout of these maps is determined by two constraints: continuity or smooth mapping of stimulus properties across the cortical surface, and coverage uniformity or uniform representation of combinations of map features over visual space. Here we use a quantitative measure of coverage uniformity (c') to test the hypothesis that cortical maps are optimized for coverage. When we perturbed the spatial relationships between ocular dominance, spatial frequency and orientation maps obtained in single regions of cortex, we found that cortical maps are at a local minimum for c'. This suggests that coverage optimization is an important organizing principle governing cortical map development.

Principal component analysis and blind separation of sources for optical imaging of intrinsic signals.

The analysis of data sets from optical imaging of intrinsic signals requires the separation of signals, which accurately reflect stimulated neuronal activity (mapping signal), from signals related to background activity. Here we show that blind separation of sources by extended spatial decorrelation (ESD) is a powerful method for the extraction of the mapping signal from the total recorded signal. ESD is based on the assumptions (i) that each signal component varies smoothly across space and (ii) that every component has zero cross-correlation functions with the other components. In contrast to the standard analysis of optical imaging data, the proposed method (i) is applicable to nonorthogonal stimulus-conditions, (ii) can remove the global signal, blood-vessel patterns, and movement artifacts, (iii) works without ad hoc assumptions about the data structure in the frequency domain, and (iv) provides a confidence measure for the signals (Z score). We first demonstrate on orientation maps from cat and ferret visual cortex, that principal component analysis, which acts as a preprocessing step to ESD, can already remove global signals from image stacks, as long as data stacks for at least two-not necessarily orthogonal-stimulus conditions are available. We then show that the full ESD analysis can further reduce global signal components and-finally-concentrate the mapping signal within a single component both for differential image stacks and for image stacks recorded during presentation of a single stimulus.

Eyes wide shut.

Crowley and Katz cast doubt on the idea that correlated activity is critical for visual cortex development by showing that ocular dominance maps can emerge without any retinal input.

Visual attention: spotlight on the primary visual cortex.

Visual search tasks appear to involve spatially selective attention to the target, but evidence for attentional modulation in the visual area with the most precise retinotopic organization V1 has been elusive. Recent imaging studies show that spatial attention can indeed enhance visual responses in human V1.

Activity-dependent regulation of alternative splicing patterns in the rat brain.

Alternative splicing plays an important role in the expression of genetic information. Among the best understood alternative splicing factors are transformer and transformer-2, which regulate sexual differentiation in Drosophila. Like the Drosophila genes, the recently identified mammalian homologues are subject to alternative splicing. Using an antibody directed against the major human transformer-2 beta isoform, we show that it has a widespread expression in the rat brain. Pilocarpine-induced neuronal activity changes the alternative splicing pattern of the human transformer-2-beta gene in the brain. After neuronal stimulation, a variant bearing high similarity to a male-specific Drosophila tra-2179 isoform is switched off in the hippocampus and is detectable in the cortex. In addition, the ratio of another short RNA isoform (htra2-beta2) to htra2-beta1 is changed. Htra2-beta2 is not translated into protein, and probably helps to regulate the relative amounts of htra2-beta1 to beta3. We also observe activity-dependent changes in alternative splicing of the clathrin light chain B, c-src and NMDAR1 genes, indicating that the coordinated change of alternative splicing patterns might contribute to molecular plasticity in the brain.

Intrinsic and environmental factors in the development of functional maps in cat visual cortex.

In the mammalian visual cortex, key neuronal response properties such as orientation preference and ocular dominance (OD) are mapped in an orderly fashion across the cortical surface. It has been known for some time that manipulating early postnatal visual experience can change the appearance of the OD map. Similar evidence for developmental plasticity of the orientation map has been scarce. We employed optical imaging of intrinsic signals to examine the contribution of intrinsic and environmental factors to the development of cortical maps, using the paradigms of strabismus, reverse occlusion and rearing in a single-orientation environment ('stripe-rearing'). For several weeks after induction of strabismus, the pattern of OD domains remained stable in young kittens. The isotropic magnification of the OD map matched the postnatal growth of the visual cortical surface during the same period. In reverse-occluded and in stripe-reared kittens, orientation preference maps obtained through the left and the right eye were very similar, although the two eyes had never shared any visual experience. We suggest that the geometry of functional maps in the visual cortex is intrinsically determined, while the relative strength of representation of different response properties can be modified through visual experience.

Spatial relationships among three columnar systems in cat area 17.

In the primary visual cortex, neurons with similar response properties are arranged in columns. As more and more columnar systems are discovered it becomes increasingly important to establish the rules that govern the geometric relationships between different columns. As a first step to examine this issue we investigated the spatial relationships between the orientation, ocular dominance, and spatial frequency domains in cat area 17. Using optical imaging of intrinsic signals we obtained high resolution maps for each of these stimulus features from the same cortical regions. We found clear relationships between orientation and ocular dominance columns: many iso-orientation lines intersected the borders between ocular dominance borders at right angles, and orientation singularities were concentrated in the center regions of the ocular dominance columns. Similar, albeit weaker geometric relationships were observed between the orientation and spatial frequency domains. The ocular dominance and spatial frequency maps were also found to be spatially related: there was a tendency for the low spatial frequency domains to avoid the border regions of the ocular dominance columns. This specific arrangement of the different columnar systems might ensure that all possible combinations of stimulus features are represented at least once in any given region of the visual cortex, thus avoiding the occurrence of functional blind spots for a particular stimulus attribute in the visual field.

Spatio-temporal frequency domains and their relation to cytochrome oxidase staining in cat visual cortex.

Spatial and temporal frequencies are important attributes of the visual scene. It is a long-standing question whether these attributes are represented in a spatially organized way in cat primary visual cortex. Using optical imaging of intrinsic signals, we show here that grating stimuli of different spatial frequencies drifting at various speeds produce distinct activity patterns. Rather than observing a map of continuously changing spatial frequency preference across the cortical surface, we found only two distinct sets of domains, one preferring low spatial frequency and high speed, and the other high spatial frequency and low speed. We compared the arrangement of these spatio-temporal frequency domains with the cytochrome oxidase staining pattern, which, based on work in primate striate cortex, is thought to reflect the partition of the visual cortex into different processing streams. We found that the cytochrome oxidase blobs in cat striate cortex coincide with domains engaged in the processing of low spatial and high temporal frequency contents of the visual scene. Together with other recent results, our data suggest that spatiotemporal frequency domains are a manifestation of parallel streams in cat visual cortex, with distinct patterns of thalamic inputs and extrastriate projections.

Guidance of thalamocortical axons by growth-promoting molecules in developing rat cerebral cortex.

Substrate-bound guidance cues play an important role during the development of thalamocortical projections. We used time-lapse video microscopy to study the growth behaviour of thalamic axons on different substrates. On embryonic cortical membranes and on a pure laminin substrate, thalamic fibres advanced relatively slowly (approximately 15 microns/h) and on average their growth cones retracted transiently every approximately 5 h. In contrast, on membranes prepared from early postnatal cortex, thalamic fibres grew twice as fast and spontaneous growth cone collapse occurred approximately 8 times less often. Experiments in which we used the sugar-binding lectin peanut agglutinin or heat inactivation to change the membrane properties indicated that these differences are due to growth-supporting molecules on postnatal cortical membranes. When offered a choice between embryonic and postnatal cortical membranes, thalamic axons preferred the postnatal membrane substrate. Time-lapse imaging revealed that borders between these two substrates effectively guided thalamic fibres, and in most cases axons changed their direction without collapse of the growth cone. Our results suggest that thalamic axons can be guided by the spatial distribution of growth-promoting molecules in the developing cortex.

Reconstructing cortical connections in a dish.

During development of the cortex, efferent projection neurons located in distinct cortical layers send their axons to different targets, and afferent fibers establish connections with cortical target cells of a particular layer. Recent studies have shown that layer- and cell-specific afferent and efferent cortical connections established in culture are similar to those observed in vivo. The results of these experiments provide evidence for the existence of diffusible and membrane-bound guidance factors for specific sets of axons. Furthermore, they suggest the use of different molecules to navigate axons towards their target, regulate target innervation and mediate target cell recognition.

Relationships between dendritic morphology and cytochrome oxidase compartments in monkey striate cortex.

In primate striate cortex, staining for the mitochondrial enzyme cytochrome oxidase reveals a regular pattern of intense staining, the blobs, which are surrounded by the lighter stained interblob regions. Neurons in both compartments exhibit profound functional differences: blob cells have color selective, unoriented receptive fields, whereas interblob cells are usually not color selective and have oriented receptive fields. Neuroanatomical tracing studies have shown that blob and interblob cells receive different inputs and participate in different projections. It is not known, however, whether this compartmental organization is also reflected in the dendritic morphology of individual cells. We therefore combined intracellular staining with cytochrome oxidase histochemistry to study the relationship between cell morphology and blob pattern in layers 2 and 3 of macaque striate cortex. Single cells were injected with the fluorescent dye lucifer yellow in lightly fixed tangential sections. Adjacent sections were reacted for cytochrome oxidase to reveal the blobs. The spatial relationship between stained cells and the pattern of the blobs were subsequently determined by aligning the sections by using radially running blood vessels as landmarks. Our results show that pyramidal cells located in blob and interblob regions do not differ in their soma size, spine density, and basal dendritic field structure. This indicates that the characteristic functional properties of the neurons in both compartments do not depend on the morphology of their dendritic trees. Since the elongation of the dendritic fields of blob and interblob cells was also found to be similar, we conclude that cortical orientation selectivity is not generated through elongated dendritic fields. We found several cells with dendrites freely crossing the borders between blob and interblob regions. These cells might correspond to cells with "mixed" receptive field properties, e.g., color selective oriented cells, which in physiological studies were found at the transition from blob to interblob regions. However, there were also a number of cells that respected the borders. A quantitative analysis of the dendritic fields revealed that 67% of the cells located close to the borders have a tendency to confine their dendrites to only one compartment. Thus the pattern of basal dendrites of these cells might be shaped by the parcellation of the striate cortex in blob and interblob regions. These dendritic field asymmetries may help to maintain the segregation at the single cell level into different processing channels in monkey striate cortex.

Morphological types of projection neurons in layer 5 of cat visual cortex.

Pyramidal cells in layer 5 of the visual cortex have multiple cortical and subcortical projection sites. Previous studies found that many cells possess bifurcating axons and innervate more than one cortical or subcortical target, but cells projecting to both cortical and subcortical targets were not observed. The present study examines the morphology of cells in cat visual cortex projecting to the superior colliculus, the main subcortical target of layer 5, and cells in layer 5 projecting to cortical areas 18 and 19. The neurons that give rise to these different projections were retrogradely labelled and intracellularly stained in living brain slices. Our results show that cells within each projection group have several morphological features in common. All corticotectal cells have a long apical dendrite forming a large terminal tuft in layer 1. Their cell bodies are medium sized to large, and their basal dendrites form a dense and symmetrical dendritic field. Corticocortical cells in layer 5 have a very different morphology: their apical dendrites are short and they never reach higher than layers 2/3. Their cells bodies are small to medium sized and they have fewer basal dendrites than corticotectal cells. Thus there are two morphologically distinct projection systems in layer 5, one projecting to cortical and the other one to subcortical targets, suggesting that these two systems transmit different information from the visual cortex. Among the corticotectal cells with the largest cell bodies we found some cells whose basal and apical dendrites were almost devoid of spines. Spiny and spinefree corticotectal cells also have different intrinsic axon collaterals and therefore play different roles in the cortical circuitry. While many spiny corticotectal cells have axon collaterals that project to layer 6, spinefree corticotectal cells have fewer axon collaterals and these do not arborize in layer 6. We suggest that the two morphological types of corticotectal cells might be related to functional differences known to exist among these cells. We discuss how the presence or absence of spines affects the integration of the synaptic input and how this might be related to the cells' functional properties.

Morphology of identified projection neurons in layer 5 of rat visual cortex.

We studied the morphology of neurons in layer 5 of rat visual cortex (area 17) projecting to the contralateral hemisphere and the superior colliculus. Double labelling with fluorescent tracers indicated that these projections arise from different populations of cells. To reveal the morphology of the cells we stained retrogradely labelled neurons intracellularly in living brain slices. Callosal projecting pyramidal cells have 3-6 basal dendrites and an apical dendrite which never reaches higher than layer 3. Corticotectal cells have 6-8 basal dendrites and a prominent apical dendrite which always forms a large tuft in layer 1. Thus, neurons in the same cortical layer that give rise to different projections also differ in their morphology. However, each population of neurons has a rather stereotyped dendritic branching pattern, despite a large variation in soma size.