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[Neural mechanisms of sound localization in owls].

Barn owls localize sound by using the interaural time difference of the horizontal plane and the interaural intensity difference for the vertical plane. The owl's auditory system possesses the two binaural cues in separate pathways in the brainstem. Owls use a process similar to cross-correlation to derive interaural time differences. Convergence of different frequency bands in the inferior colliculus solves the problems of phase-ambiguity which is inherent in cross-correlating periodic signals. The two pathways converge in the external nucleus of the inferior colliculus to give rise to neurons that are selective for combinations of the two cues. These neurons form a map of auditory space. The map projects to the optic tectum to form a bimodal map which, in turn, projects to a motor map for head turning. The visual system calibrates the auditory space map during ontogeny in which acoustic variables change. In addition to this tectal pathway, the forebrain can also control the sound-localizing behaviour.

Animals↗

Experimental test of a perturbation model for time-resolved imaging in diffusive media.

We report what to our knowledge is a novel perturbation approach for time-resolved transmittance imaging in diffusive media, based on the diffusion approximation with extrapolated boundary conditions. The model relies on the method of Padé approximants and consists of a nonlinear approximation of time-resolved transmittance curves in the presence of an inclusion. The proposed model is intended to extend the range of applicability of perturbation models when applied to inclusions that are non-point-like. We test the model on different tissue phantoms with scattering only, absorbing only, and both scattering and absorbing inclusions. Maps of the optical properties are displayed, and the results are compared with those obtained by means of the usual linear approximation of time-resolved transmittance curves. We found that the nonlinear approach gives a better prediction for absolute values of the scattering and absorption coefficients of inclusions, when the inclusion optical properties are higher than the surrounding background. Furthermore, better-resolved spots and a reduced cross talk between the two parameters are found in the reconstructed maps. Because the range of the optical properties spanned by the considered phantoms covers the values expected for optical mammography, the application of the reported reconstruction method to in vivo images of a breast appears promising from a diagnostic viewpoint.

Biological Phenomena↗

In vivo monitoring of cutaneous edema using spectral imaging in the visible and near infrared.

Tissue inflammation is often accompanied by local interstitial fluid accumulation expressed as edema. Edema can be the manifestation of infection, lymphatic blockage, wound healing, or even cancer, and is typically graded visually. Here we demonstrate that the edema reaction can be objectively quantitated in vivo by the use of spectral imaging. To this end we applied the method on a histamine-induced cutaneous edema model. Apparent concentrations of oxy-hemoglobin, deoxy-hemoglobin, and water were calculated for each pixel of a spectral image stack. These values were used to construct concentration maps for each of these molecules as well as an intensity map of an optical tissue-scattering parameter. The oxy-hemoglobin and the tissue water maps are two-dimensional quantitative representations of the skin areas involved in erythema and edema, respectively. These maps demonstrated characteristics of the wheal-and-flare reaction and their gray-level intensities were dependent on the applied histamine dose. We conclude that spectral imaging can be a valuable noninvasive tool in the study of edema pathology and can be used to monitor the edema reaction in vivo or follow the efficacy of treatments in a clinical setting.

Adult↗

Representation of the temporal raphe within the optic tract of the cat.

The retinal topography of the cat's optic tract was determined by means of injections of the enzyme horseradish peroxidase (HRP) into the tract. This analysis was accomplished by the subtraction of all HRP injection sites not labeling a defined retinal area from those injection sites which resulted in ganglion cell labeling (Venn diagram analysis). Using this method, the following correspondences were demonstrated for the ipsilateral and contralateral projections: superior retina represented in medial optic tract; inferior retina in lateral tract; and area centralis in a dorsocentral location (which was part of a larger area representing the visual streak). The temporal raphe was represented in the ipsilateral tract as a band curving from the area centralis region toward the dorsomedial border of the tract. Contralateral fibers from a region superior to the optic disc were found to be displaced with respect to the general retinal representation in the optic tract and this appeared to be related to retinal development. The ratio of contralateral to ipsilateral fibers was determined and found to be nonuniform within the tract. Injection of HRP into the optic tract of the cat also allowed the axons from labeled retinal ganglion cells to be traced within the retina and optic disc. Axons from ganglion cells lying temporal to the raphe curve around the area centralis enter the optic disc on the lateral and inferior aspects. Ganglion cells lying nasal to the raphe send their axons more directly to enter the optic disc on its superior aspect. A schema is proposed whereby the retina is mapped onto the optic tract.

Animals↗

Optically active benzamides as predictive tools for mapping the dopamine D2 receptor.

Substituent variations on the pyrrolidinyl nitrogen of sulpiride, a selective D2 dopamine antagonist, showed that in vitro and in vivo activities are concentrated in the (S) optical series for N-alkyl analogs and in the (R) series for N-benzyl analogs. To account for these unusual structure-activity relationships, a pharmacophoric model was built from the crystallographic structure of (-)piquindone and extended to 14 other D2 antagonists. This model considers the lone pair orientation of the basic nitrogen rather than its spatial location. Two distinct active conformations for benzamides were defined, corresponding to the (S) and (R) series. An extended pharmacophore is then proposed involving four main anchoring areas: (i) an aromatic site Ar1, (ii) a tertiary nitrogen with its lone pair orthogonal to the Ar1 plane, (iii) a dipole delta 1 coplanar to the Ar1 ring and (iv) three sites for the N-substituent, including a small hydrophobic pocket and two different aromatic binding sites Ar2 and Ar3. To probe the predictive value of this model, structures were designed and several compounds were synthesized and tested as inhibitors of [125I]iodo-sulpiride binding to rat striatal membranes and as antagonists of apomorphine-induced stereotyped behavior in mice.

Animals↗

Synthesis, axonal transport, and turnover of the high molecular weight microtubule-associated protein MAP 1A in mouse retinal ganglion cells: tubulin and MAP 1A display distinct transport kinetics.

Microtubule-associated proteins (MAPs) in neurons establish functional associations with microtubules, sometimes at considerable distances from their site of synthesis. In this study we identified MAP 1A in mouse retinal ganglion cells and characterized for the first time its in vivo dynamics in relation to axonally transported tubulin. A soluble 340-kD polypeptide was strongly radiolabeled in ganglion cells after intravitreal injection of [35S]methionine or [3H]proline. This polypeptide was identified as MAP 1A on the basis of its co-migration on SDS gels with MAP 1A from brain microtubules; its co-assembly with microtubules in the presence of taxol or during cycles of assembly-disassembly; and its cross-reaction with well-characterized antibodies against MAP 1A in immunoblotting and immunoprecipitation assays. Glial cells of the optic nerve synthesized considerably less MAP 1A than neurons. The axoplasmic transport of MAP 1A differed from that of tubulin. Using two separate methods, we observed that MAP 1A advanced along optic axons at a rate of 1.0-1.2 mm/d, a rate typical of the Group IV (SCb) phase of transport, while tubulin moved 0.1-0.2 mm/d, a group V (SCa) transport rate. At least 13% of the newly synthesized MAP 1A entering optic axons was incorporated uniformly along axons into stationary axonal structures. The half-residence time of stationary MAP 1A in axons (55-60 d) was 4.6 times longer than that of MAP 1A moving in Group IV, indicating that at least 44% of the total MAP 1A in axons is stationary. These results demonstrate that cytoskeletal proteins that become functionally associated with each other in axons may be delivered to these sites at different transport rates. Stable associations between axonal constituents moving at different velocities could develop when these elements leave the transport vector and incorporate into the stationary cytoskeleton.

Animals↗

Mapping and ultrastructure of serotonin-immunoreactive neurons in the optic lobes of three insect species.

With antibodies to serotonin (5-HT) we have mapped immunoreactive neurons in the optic lobes of three species, the blowfly Calliphora, the desert ant Cataglyphis, and the worker bee Apis. The main emphasis in this investigation is on a system of 5-HT-positive neurons connecting the most peripheral neuropil of the optic lobes, the lamina, to more central neuropil regions. To aid in electron microscopical identification of these neurons we used immunocytochemistry at the EM-level and Golgi-EM for Calliphora and horseradish peroxidase (HRP) labelling for the other two insects. The immunoreactive terminals in Calliphora and the HRP-labelled ones in the other insects contain large (c. 100 nm) granular vesicles and smaller (c.60 nm) clear vesicles. In Cataglyphis and Apis the profiles with granular vesicles are presynaptic to second order neurons of the lamina, whereas in Calliphora no synaptic contacts were found. In this animal the 5-HT-positive terminals are situated distal to the synaptic layer of the lamina, in a region of retinal photoreceptor axons and perikarya of the lamina monopolar neurons. In Catagylphis and Apis the interactions of the 5-HT-neurons with the laminar neurons might occur through chemical synapses, whereas in Calliphora neuroactive substance could be released non-synaptically from varicosities distal to the synaptic layer. The possible involvement of 5-HT in control of neuronal activity in the optic lobes is discussed.

Animals↗

Simultaneous recording of event-related auditory oddball response using transcranial near infrared optical topography and surface EEG.

Near infrared optical topography (OT) is the measurement of hemoglobin absorption simultaneously from an array of optical fibers on the scalp to construct maps of cortical activity. We demonstrate that OT can be used to simultaneously detect and characterize the hemodynamic responses associated with an "oddball" auditory stimulus and that corresponding electrical event related potentials can be acquired simultaneously using conventional scalp recordings. In addition to the measured electrical response, the hemodynamic localization is consistent with fMRI studies, which show significant activation in the temporal and parietal cortical regions. The event-related response of total hemoglobin showed relatively slow peak latencies (5.8 +/- 0.3 s), which were also consistent with fMRI. The current study shows the regions of peak hemodynamic activity that are in closest proximity to areas of peak electrical activity. This is the first demonstration of simultaneous ERP electrical recording and non-invasive optical mapping in human subjects, which promises to be an important tool in the characterization of both normal and abnormal brain function.

Auditory Cortex↗

Expression of glutamate receptor genes in white matter: developing and adult rat optic nerve.

We have identified the glutamate receptor genes expressed by developing and adult rat optic nerve. Results from PCR suggested that of the ionotropic glutamate receptor (GluR) gene family, only GluR1 and GluR3 subunits are expressed by optic nerve. However, Northern blot analysis demonstrated that only the GluR1 subtype is expressed at appreciable levels. In situ hybridization histochemistry of GluR1 was performed and a diffuse pattern of expression was observed in both postnatal day 15 and adult optic nerve. Restriction mapping of the GluR1 PCR product indicated the expression of the flip alternative splice version in optic nerve. The detected GluR1 message was not due to axons because no loss of expression was observed after the degeneration of axons. PCR analysis also revealed the presence of a metabotropic glutamate receptor in optic nerve.

Aging↗

Vesikin, a vesicle associated ATPase from squid axoplasm and optic lobe, has characteristics in common with vertebrate brain MAP 1 and MAP 2.

Vesikin, a protein that can associate with squid axoplasmic vesicles or optic lobe microtubules, has been implicated as a force-generating molecule involved in microtubule-dependent vesicle transport [Gilbert and Sloboda, 1986, 1988]. Because vesikin crossreacts with an antibody to porcine brain microtubule associated protein 2 (MAP 2), studies were conducted to compare squid vesikin and brain MAPs. When taxol stabilized microtubules containing vesikin as a microtubule associated protein were incubated in the presence of ATP, vesikin dissociated from the microtubule subunit lattice. This behavior would be expected for an ATP-dependent, force generating molecule that serves as a crossbridge between vesicles and microtubules. When chick brain microtubules were treated under the same conditions, MAP 2 remained bound to the microtubules while MAP 1 dissociated in a manner similar to vesikin. One dimensional peptide mapping procedures revealed that, although digestion of vesikin and MAP 2 generated several peptides common to both proteins, vesikin and MAP 2 are clearly not identical. Furthermore, the addition of vesikin or MAPS 1 and 2 to purified tubulin stimulated microtubule assembly in a manner dependent on the concentration of added protein. These findings demonstrate that brain MAPs share characteristics common to squid vesikin and support the suggestion that brain MAPs 1 and 2 might act as a force generating complex for vesicle transport in higher organisms.

Adenosine Triphosphate↗

Clinical observations on the isolated optic nerve injury.

The optic nerve can be injured indirectly by blunt trauma to the frontoorbital region. The mechanism of injury is related to trauma of the nerve in the tight, bony optic canal. Neurovisual transmission in the canal is interrupted by pressure on the nerve. The conventional management includes careful observation and supportive treatment. Recent interest in this unique clinical problem has prompted the development of new techniques to decompress the injured optic canal extracranially; these approaches have produced encouraging results. Another dividend of the method has been the mapping of the optic nerve region under magnification using the operating room microscope. Any gain of vision, partial or complete, is well appreciated by the patient and the physician.

Craniocerebral Trauma↗

Ultraviolet illumination thermoreflectance for the temperature mapping of integrated circuits.

Noncontact optical methods such as thermoreflectance, which measure temperature-induced optical reflectivity changes, are particularly suitable for obtaining high-resolution temperature mappings on integrated circuits. Unfortunately, the coefficient linking the variations of temperature and reflectivity depends on the nature of the material and can be modified when optical interferences occur in the Si3N4-based encapsulation layers protecting the circuits. We show that taking advantage of the deep UV absorption of encapsulation layers yields temperature mapping that is independent of the underlying materials. A single calibration is therefore enough to yield the temperature on any point of the uniform and thermally thin encapsulation layer. This simplification and its potential for high resolution should make UV thermoreflectance more attractive to the semiconductor industry.

Journal Article↗

A novel optical imaging method for the early detection, quantitative grading, and mapping of cancerous and precancerous lesions of cervix.

This paper describes a novel optical imaging method for the in vivo early detection, quantitative staging, and mapping of cervical cancer and precancer. A multispectral imaging system was developed, which is capable of performing time-resolved imaging spectroscopy. The system was used in order to assess quantitatively the alterations in the light scattering properties of the cervix, induced selectively and reversibly in cervical neoplasias, after the application of acetic acid solution. Spectral imaging and analysis of cervix show that the maximum contrast between acetic acid responsive and nonresponsive areas is obtained at 525 +/- 15 nm, which is further enhanced by cutting off the regular component of tissue reflection, with the aid of two linear cross polarizers. Successive snapshot imaging at this spectral band enables the quantitative assessment of the temporal alterations in the intensity of the backscattered light, in any spatial location of the examined area. Initial clinical trials show that optical contrast enhancement results in a notable improvement of the sensitivity in detecting incipient lesions. It was also shown that the measured temporal characteristics of the phenomenon contain specific information, which enables the differentiation between neoplastic and nonneoplastic lesions, as well as between neoplasias of different grade. The demonstrated improved sensitivity and specificity highlight the potential of the method in both clinical research and noninvasive diagnosis.

Acetic Acid↗

Mapping retinotopic structure in mouse visual cortex with optical imaging.

We have used optical imaging of intrinsic signals to visualize the retinotopic organization of mouse visual cortex. The functionally determined position, size, and shape of area 17 corresponded precisely to the location of this area as seen in stained cortical sections. The retinotopic map, which was confirmed with electrophysiological recordings, exhibited very low inter-animal variability, thus allowing averaging of maps across animals. Patches of activity in area 17 were often encircled by regions in which the intrinsic signal dropped below baseline, suggesting the presence of strong surround inhibition. Single-unit recordings revealed that this decrease of the intrinsic signal indeed correlated with a drop of neuronal firing rate below baseline. The averaged maps also greatly facilitated the identification of extrastriate visual activity, pointing to at least four extrastriate visual areas in the mouse. We conclude that optical imaging is ideally suited to visualize retinotopic maps in mice, thus making this a powerful technique for the analysis of map structure in transgenic animals.

Animals↗

Depth profiling of the optical absorption coefficient in ultraviolet-degraded poly(vinylchloride) films by dual beam light profile microscopy.

Dual beam laser light profile microscopy (LPM) was applied in this work to the depth mapping of the optical absorption coefficient in photo-degraded poly(vinyl chloride) films. Depth profiles followed the absorption coefficient of a conjugated polyene photoproduct at visible wavelengths in photolyzed films of approximately 200 mum thickness. Both continuous and layered (separable laminate) films were studied. The absorption coefficient profiles reconstructed from photo-degraded thin films showed the classic concentration profiles seen in the literature for PVC degraded in the presence of oxygen and nitrogen atmospheres. In the case of single thin layers with continuous properties, the depth profiles were smooth and regular with minimum spatial noise. In the LPM of laminate structures, more optical anomalies were present because of the multiply interfacial structures that appeared in both the images and the reconstructed depth profiles. Notwithstanding, it was possible to profile the optical absorption coefficient at a level of error comparable to standard microtome methods. The latter was determined by comparing the LPM results to a destructive layer-by-layer analysis performed in parallel on the imaged materials. The dual beam LPM method should be generally useful for establishing polyene concentration profiles in industrial materials produced by photochemical, thermal, and chemical degradation mechanisms.

Absorption↗

Evidence that regenerating optic axons maintain long-term growth in the lizard Ctenophorus ornatus: growth-associated protein-43 and gefiltin expression.

In the lizard, Ctenophorus ornatus, the optic nerve regenerates but animals remain blind via the experimental eye, presumably as a result of axons failing to consolidate a retinotopic map in the optic tectum. Here we have examined immunohistochemically the expression of the growth-associated protein GAP-43 and the low-molecular-weight intermediate filament protein gefiltin, up to one year after optic nerve crush. Both proteins were found to be permanently up-regulated, suggesting that regenerating axons are held in a permanent state of re-growth. We speculate that, in the lizard, the continued expression of GAP-43 and the failure to switch from the expression of low- to high-molecular-weight intermediate filament proteins are associated with the inability to consolidate a retinotopic projection.

Animals↗