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Area and layer patterning in the developing cerebral cortex.

Two anatomical patterns characterize the neocortex, and both are essential for normal cortical function. First, neocortex is divided into anatomically distinct and functionally specialized areas that form a species-specific map. Second, neocortex is composed of layers that organize cortical connectivity. Recent studies of layer and area development have used time-lapse microscopy to follow cortical cell division and migration, gene arrays to find layer- or area- specific regulatory genes, time- and region- specific manipulations of candidate genes, and optical imaging to compare area maps in wild type with genetically altered mice. New observations clarify the molecular and cellular mechanisms that generate each pattern, and stress the links between layer and area formation.

Animals↗

Clinical applications of the Shack-Hartmann aberrometer.

The efficacy of the Shack-Hartmann technique for measuring the optical aberrations of the eye was evaluated for four classes of clinical conditions associated with optically abnormal eyes. These categories (with specific examples) are: anomalies of the tear film (dry eye), corneal disease (keratoconus), corneal refractive surgery [laser-assisted in situ keratomileusis (LASIK)], and lenticular cataract. We show that in each of these cases, it is possible to obtain at least a partial topographic map of the refractive aberrations of the patient's eyes, but severe losses of data integrity can occur. We further show that the Shack-Hartmann aberrometer provides additional information about the eye's imperfections on a very fine spatial scale (< 0.4 mm) which scatter light and further degrade the quality of the retinal image. Taken together, spatial maps of the variation of optical aberrations and scatter across the eye's entrance pupil represents an improved description of the optical imperfections of the abnormal eye.

Cataract↗

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.

Animals↗

Optical imaging of the retinotopic organization of V1 in the common marmoset.

We examined the retinotopic mapping of the visual world in the primary visual cortex of the marmoset monkey using differential optical imaging. Two sets of complementary stripe-like locations were visually stimulated in turn. Their difference depicts the cortical representations of continuous bands of visual space. By rotating the sets of stripe-like locations it is possible to map different spatial axes. Analogous to the macaque we found that the V1/V2 border represented the vertical meridian, while horizontal, 45-, and 135-degree angled stripes of space were also represented in a continuous manner. We developed a new automatic method of calculating local measures of cortical magnification from our optical retinotopic maps. Using this method we found no evidence of any local anisotropies in cortical representation. Overall our results indicate that space is mapped isotropically in the primary visual cortex of the common marmoset.

Algorithms↗

Changes in fiber order in the optic nerve and tract of rat embryos.

In order to define the extent to which retinotopic order in the optic pathways may contribute to fiber segregation at the chiasm or to the formation of central maps, the arrangement of fibers in the optic nerve and tract of rat embryos, on embryonic days 16.5 and 18.5, has been studied by placing a small granule of DiI in one of the four quadrants of the retina and tracing the filled fibers through transverse sections of the retinofugal pathway with confocal microscopy. There is a distinct quadrant-specific order in the optic stalk immediately behind the eye, with fibers from the ventral nasal, dorsal nasal, dorsal temporal, and ventral temporal retina arranged sequentially across the rostrocaudal axis of the cross section of the stalk. However, this distinct order is not maintained very far. There is a gradual increase in the degree of overlap between fibers from the different quadrants as the fibers pass towards the chiasm. The dorsal groups of fibers intermingle extensively along almost the entire length of the stalk, but the fibers from ventral sectors remain separate until they reach the prechiasmatic region, where the ventral temporal and the ventral nasal fibers spread throughout the rostrocaudal extent of the stalk and the chiasm. The initial quadrant-specific order is completely lost at the chiasm. However, beyond the optic chiasm, the fibers are reorganized into another distinct order. In the optic tract, there is a segregation of dorsal from ventral fibers, but the nasal and temporal groups remain intermingled. The results of this study indicate that the earliest fibers in the developing optic tract are arranged according to topographical rules that differ from those obtaining behind the eye. Since all topographical order is lost between these two levels, there must be an active sorting mechanism in the region where the chiasm joins the tract. Possibly this mechanism is related to the development of the dorsoventral axis of the topographic maps in the central visual targets.

Animals↗

Molecular mechanisms of optic axon guidance.

Axon guidance is one of the critical processes during vertebrate central nervous system (CNS) development. The optic nerve, which contains the axons of retinal ganglion cells, has been used as a powerful model to elucidate some of the mechanisms underlying axon guidance because it is easily manipulated experimentally, and its function is well understood. Recent molecular biology studies have revealed that numerous guidance molecules control the development of the visual pathway. This review introduces the molecular mechanisms involved in each critical step during optic axon guidance. Axonal projections to the optic disc are thought to depend on adhesion molecules and inhibitory extracellular matrices such as chondroitin sulfate. The formation of the head of the optic nerve and the optic chiasm require ligand-receptor interactions between netrin-1 and the deleted in colorectal cancer receptor, and Slit proteins and Robo receptors, respectively. The gradient distributions of ephrin ligands and Eph receptors are essential for correct ipsilateral projections at the optic chiasm and the topographic mapping of axons in the superior colliculus/optic tectum. The precise gradient is regulated by transcription factors determining the retinal dorso-ventral and nasal-temporal polarities. Moreover, the axon guidance activities by Slit and semaphorin 5A require the existence of heparan sulfate, which binds to numerous guidance molecules. Recent discoveries about the molecular mechanisms underlying optic nerve guidance will facilitate progress in CNS developmental biology and axon-regeneration therapy.

Animals↗

Complementary postsynaptic activity patterns elicited in olfactory bulb by stimulation of mitral/tufted and centrifugal fiber inputs to granule cells.

Main olfactory bulb (MOB) granule cells receive spatially segregated glutamatergic synaptic inputs from the dendrites of mitral/tufted cells as well as from the axons of centrifugal fibers (CFFs) originating in olfactory cortical areas. Dendrodendritic synapses from mitral/tufted cells occur on granule cell distal dendrites in the external plexiform layer (EPL), whereas CFFs preferentially target the somata/proximal dendrites of granule cells in the granule cell layer (GCL). In the present study, tract tracing, and recordings of field potentials and voltage-sensitive dye optical signals were used to map activity patterns elicited by activation of these two inputs to granule cells in mouse olfactory bulb slices. Stimulation of the lateral olfactory tract (LOT) produced a negative field potential in the EPL and a positivity in the GCL. CFF stimulation produced field potentials of opposite polarity in the EPL and GCL to those elicited by LOT. LOT-evoked optical signals appeared in the EPL and spread subsequently to deeper layers, whereas CFF-evoked responses appeared in the GCL and then spread superficially. Evoked responses were reduced by N-methyl-d-aspartate (NMDA) receptor antagonists and completely suppressed by AMPA receptor antagonists. Reduction of extracellular Mg(2+) enhanced the strength and spatiotemporal extent of the evoked responses. These and additional findings indicate that LOT- and CFF-evoked field potentials and optical signals reflect postsynaptic activity in granule cells, with moderate NMDA and dominant AMPA receptor components. Taken together, these results demonstrate that LOT and CFF stimulation in MOB slices selectively activate glutamatergic inputs to the distal dendrites versus somata/proximal dendrites of granule cells.

Action Potentials↗

Subwavelength focusing and guiding of surface plasmons.

The constructive interference of surface plasmon polaritons (SPP) launched by nanometric holes allows us to focus SPP into a spot of high near-field intensity having subwavelength width. Near-field scanning optical microscopy is used to map the local SPP intensity. The resulting SPP patterns and their polarization dependence are accurately described in model calculations based on a dipolar model for the SPP emission at each hole. Furthermore, we show that the high SPP intensity in the focal spot can be launched and propagated on a Ag strip guide with a 250 x 50 nm2 cross section, thus overcoming the diffraction limit of conventional optics. The combination of focusing arrays and nano-waveguides may serve as a basic element in planar nano-photonic circuits.

Equipment Design↗

Receptive fields of opponent color units in the optic nerve of the ground squirrel.

When mapped with white light, each receptive field consisted either of an excitatory ("on") center and a concentric inhibitory ("off") surround, or of the reverse arrangement. Monochromatic stimuli revealed that each receptive field was composed of two mutually antagonistic components (one excitatory, one inhibitory) which had different spectral sensitivities and different spatial distributions. For some units the two chromatic components had identical spatial distributions.

Animals↗

Wallerian degeneration in the optic radiation after temporal lobectomy demonstrated in vivo with diffusion tensor imaging.

PURPOSE: Diffusion tensor imaging allows the quantitative assessment of the microstructural organization of tracts in vivo (MR tractography). We used the new technique of MR tractography to demonstrate the effects of temporal lobectomy on the optic radiation. METHODS: Spatially normalised maps encoding magnitude of the bias (anisotropy) of diffusion of three patients with temporal lobe resections were compared with spatially normalised diffusion maps of 22 control subjects. All three patients were operated on for the treatment of medically intractable temporal lobe epilepsy and had a normal neurologic examination before surgery. One patient had an amygdalocorticectomy. Two patients had standard en bloc resections, one of whom developed a homonymous hemianopia after surgery. RESULTS: In the patient with hemianopia, a significant reduction of diffusion anisotropy (greater than mean+/-2 SD) consistent with wallerian degeneration was demonstrated in the optic radiation on the side of the temporal lobectomy, extending from the temporal to the occipital lobe. In the other patient with standard en bloc resection but clinically no hemianopia, the optic radiation was only marginally affected. In the third patient (amygdalocorticectomy), the diffusion anisotropy was within the normal range in the expected position of the optic radiation. CONCLUSIONS: Our findings show that MR tractography may be a useful tool to demonstrate wallerian degeneration in the optic radiation after temporal lobectomy in patients with hemianopia. This is the first time that this new method has been applied in postoperative imaging; it enables us to visualise the morphologic correlate of dysfunctional pathways after epilepsy surgery in vivo. The potential for using MR tractography to study other aspects of epilepsy is discussed.

Anisotropy↗

Optical imaging of epileptiform activity in human neocortex.

The surgical outcomes of patients suffering from neocortical epilepsy are not as successful as the surgical outcomes from resections of epilepsy patients with mesial temporal sclerosis. The main difficulty in the treatment of neocortical epilepsy is that current technology has limited accuracy in mapping neocortical epileptogenic tissue. It is known that the optical spectroscopic properties of brain tissue are correlated with changes in neuronal activity. The method of mapping these activity-evoked optical changes is known as imaging of intrinsic optical signals (IIOS). Activity-evoked optical changes measured in neocortex are generated by changes in cerebral hemodynamics (i.e., changes in blood oxygenation and blood volume). Our experimental approach was to acquire high-resolution IIOS maps of epileptiform activity in patients undergoing surgery for medically intractable neocortical epilepsy. Both spontaneous and stimulation-evoked epileptiform activity was monitored. Imaging of intrinsic optical signals was able to localize neocortical epileptic foci precisely by using changes in blood volume in contrast to changes in blood oxygenation. IIOS has the potential to translate from a purely research tool to a new intraoperative approach for the surgical treatment of neocortical epilepsy.

Blood Volume↗

Ray vector fields, prismatic effect, and thick astigmatic optical systems.

The application of the concept of ray vector fields to optical systems is reexamined. Paraxial or linear optics defines a four-dimensional ray vector field for any optical system: the vector field maps the incident ray vector into the emergent ray vector. In the case of thin systems, including thin astigmatic lenses, one can define a vector field of reduced dimensionality: the vector field is two-dimensional and maps the ray's incident position into the change in reduced direction. When the index of refraction is the same before and after a thin system, the change in reduced direction is the reduced deflection through the system or the reduced prismatic effect. Contrary to what has recently been claimed, this type of two-dimensional vector field does not apply in general to thick systems. However, a number of different types of two-dimensional vector fields can be defined for various particular classes of optical systems. Thick systems differ qualitatively from thin systems. They do not have equivalent thin lenses and cannot generally be replaced by thin lenses. Equations are derived for the change in reduced direction and deflection for a ray through optical systems in general and through separated two- and three-lens systems in particular.

Astigmatism↗

An electrophysiological study of early retinotectal projection patterns during regeneration following optic nerve crush inside the cranium in Hyla moorei.

The sequence of regeneration following intracranial optic nerve crush has been studied using electrophysiological visual mapping in the frog Hyla moorei. Compared to our earlier series with extracranial crush, the time course was slower and the intermediate projections more disorganized. It is suggested that the apparent discrepancy between the early patterns of regeneration in Xenopus and Rana compared to Hyla, fish and newt, is not due to species differences but to the location of the lesion site.

Animals↗

The orientation of the visuotectal map in Xenopus: developmental aspects.

Rotations and translocations of the eye anlage were performed in Xenopus embryos of stages ranging from 21/22 to 30. Some of the operations involved grafting wild-type eye anlagen into albino host orbits. Operations were performed under a variety of operating media and conditions. In later larval life, or after metamorphosis, the visuotectal maps from the operated eyes were recorded electrophysiologically. Results fell into two classes. In the majority, the orientation of the visuotopic map corresponded to the orientation of the eye at the time of recording. In the minority the visuotopic maps were 'compound', consisting of two parts each with its own independent orientation. The organization of the compound maps was such that one component was oriented in correspondence with the orientation of the eye, while the other component was normally oriented. Histological analysis and observations on genetically marked grafts indicated that the component parts of the compound eye were of dual cellular origin. The component giving the rotated (or translocated) map belonged to the originally operated eye tissue; whereas the component giving the normally oriented map was derived from newly grown eye tissue coming from the optic stalk. In no case was a normally oriented map obtained from a rotated or translocated eye. The results are discussed in relation to mechanisms proposed to account for the determination of map-related retinal specificity.

Animals↗

Anatomical and functional imaging of neurons using 2-photon laser scanning microscopy.

Light scattering by brain tissue and phototoxicity are major obstacles to the use of high-resolution optical imaging and photo-activation ('uncaging') of bioactive compounds from inactive ('caged') precursors in intact and semi-intact nervous systems. Optical methods based on 2-photon excitation promise to reduce these obstacles (Denk, 1994; Denk et al., 1990, 1994). Here we show a range of imaging modes based on 2-photon laser scanning microscopy (TPLSM) as applicable to problems in neuroscience. Fluorescence images were taken of neurons labeled with ion-sensitive and voltage-sensitive dyes in invertebrate ganglia, mammalian brain slices, and from the intact mammalian brain. Scanning photochemical images with whole-cell current detection (Denk, 1994) show how the distribution of neurotransmitter receptors on the surface of specific cells can be mapped. All images show strong optical sectioning and usable images can be obtained at depths greater than 100 microns below the surface of the preparation.

Animals↗

Evoked response potential markers for anesthetic and behavioral states.

The rodent whisker sensory system is a commonly used model of cortical processing; however, anesthetics cause profound differences in the shape and timing of evoked responses. Evoked response studies, especially those that use spatial mapping techniques, such as fMRI or optical imaging, will thus show significantly different results depending on the anesthesia used. To describe the effect of behavioral states and commonly used anesthetics, we characterized the early surface-evoked response potentials (ERPs) components (first ERP peak: gamma band 25-45 Hz; fast oscillation: 200-400 Hz; and very fast oscillation: 400-600 Hz) using a 25-channel electrode array on the somatosensory cortex during whisker stimulation. We found significant differences in the ERP shape when ketamine/xylazine, urethane, propofol, isoflurane, and pentobarbital sodium were administered and during sleep and wake states. The highest ERP amplitudes were observed under propofol anesthesia and during quiet sleep. Under isoflurane, the ERP was nearly absent, except for a very late component, which was concombinant with burst synchronization. The slowest responses were seen under urethane and propofol anesthesia. Spatial mapping experiments that use electrical, NMR, or optical techniques must consider the anesthetic dependency of these signals, especially when stimulation protocols or electrical and metabolic responses are compared.

Anesthesia, General↗

Three-dimensional retinal imaging with high-speed ultrahigh-resolution optical coherence tomography.

PURPOSE: To demonstrate high-speed, ultrahigh-resolution, 3-dimensional optical coherence tomography (3D OCT) and new protocols for retinal imaging. METHODS: Ultrahigh-resolution OCT using broadband light sources achieves axial image resolutions of approximately 2 microm compared with standard 10-microm-resolution OCT current commercial instruments. High-speed OCT using spectral/Fourier domain detection enables dramatic increases in imaging speeds. Three-dimensional OCT retinal imaging is performed in normal human subjects using high-speed ultrahigh-resolution OCT. Three-dimensional OCT data of the macula and optic disc are acquired using a dense raster scan pattern. New processing and display methods for generating virtual OCT fundus images; cross-sectional OCT images with arbitrary orientations; quantitative maps of retinal, nerve fiber layer, and other intraretinal layer thicknesses; and optic nerve head topographic parameters are demonstrated. RESULTS: Three-dimensional OCT imaging enables new imaging protocols that improve visualization and mapping of retinal microstructure. An OCT fundus image can be generated directly from the 3D OCT data, which enables precise and repeatable registration of cross-sectional OCT images and thickness maps with fundus features. Optical coherence tomography images with arbitrary orientations, such as circumpapillary scans, can be generated from 3D OCT data. Mapping of total retinal thickness and thicknesses of the nerve fiber layer, photoreceptor layer, and other intraretinal layers is demonstrated. Measurement of optic nerve head topography and disc parameters is also possible. Three-dimensional OCT enables measurements that are similar to those of standard instruments, including the StratusOCT, GDx, HRT, and RTA. CONCLUSION: Three-dimensional OCT imaging can be performed using high-speed ultrahigh-resolution OCT. Three-dimensional OCT provides comprehensive visualization and mapping of retinal microstructures. The high data acquisition speeds enable high-density data sets with large numbers of transverse positions on the retina, which reduces the possibility of missing focal pathologies. In addition to providing image information such as OCT cross-sectional images, OCT fundus images, and 3D rendering, quantitative measurement and mapping of intraretinal layer thickness and topographic features of the optic disc are possible. We hope that 3D OCT imaging may help to elucidate the structural changes associated with retinal disease as well as improve early diagnosis and monitoring of disease progression and response to treatment.

Anatomy, Cross-Sectional↗

Near-IR imaging of atheromas in living arterial tissue.

A near-IR imaging system and parallel vector supercomputer are used with a fiber-optic probe to produce chemical maps of the intimal surface of living arteries. Spectrometric information collected at hundreds of near-IR wavelengths is assembled into color pictures of the lipoprotein and apolipoprotein composition of atheromas using a vectorized 3-D cellular automaton-based algorithm that operates in parallel. The nonparametric mathematics developed to identify and quantify the constituents of each voxel in the artery wall avoid the matrix factorizations that generate excess error in other pattern recognition methods and permit analysis in a wavelength space of over 1000 dimensions using fewer than 100 calibration samples. A surface feature resolution of 5.5 microns and depth resolution of 6.5 microns are achieved with the system. Data from the fiber optics confirm the injury hypothesis of lesion formation and the differing roles of HDL and LDL in cholesterol transport. In clinical studies, approximately 1/2 of human arterial lesions appear fibrous and contain little or no lipid. As such, these lesions would not be expected to regress in response to cholesterol-lowering agents such as lovastatin. Identification of lesion types in vivo will enhance the efficacy of treatment programs.

Apolipoproteins↗