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Optical imaging of low Mg(2+)-induced spontaneous epileptiform activity in combined rat entorhinal cortex-hippocampal slices.

A reproducible increase in transmission of infrared light was observed during spontaneous seizure-like events (SLEs) induced by low Mg2+ solutions in combined rat entorhinal cortex-hippocampal slices. Comparison of half maxima of transmission change in different regions indicated propagation of SLEs from the medial entorhinal cortex towards the temporal cortex suggesting spread along existing anatomical pathways. Thus, optical imaging of spontaneous epileptiform activity is possible and may improve the assessment of spread patterns. The optical signal outlasted both SLEs and associated K+ signals. In contrast, the tetraethylammonium signal, indicating changes of the extracellular space (ECS) volume, had a longer time course than the transmission changes. ECS volume changes are widely held to be responsible for transmission change. Our data suggest that other mechanisms may also contribute to increased light transmission during epileptiform activity.

Animals↗

Areal extent quantification of functional representations using intrinsic signal optical imaging.

An important parameter often investigated in the characterization of cortical functional organization is the areal extent of functional modules. Because it allows the visualization of functional modules with high spatial resolution in a noninvasive way to the cortex, intrinsic signal optical imaging (ISI) can be employed for the quantification of these areal extents. The present paper describes the use of the normalized threshold analysis of areal extent quantification for the objective assessment of single-whisker functional representations in the primary somatosensory cortex of adult rats. As the success of areal extent quantification depends on the ability of ISI to allow visualization of cortical representations with minimal stimulus-dependent blood vessel representations, which are commonly encountered by ISI, the present paper also describes the further development of the intratrial analysis of visualization for minimizing these vessel representations. Both analyses are discussed with respect to their advantages as well as their inherent limitations.

Animals↗

Optical imaging of long-lasting depolarization on burst stimulation in area CA1 of rat hippocampal slices.

Postsynaptic depolarization of dendrites paired with spike generation at the soma is considered to be a central mechanism of long-term potentiation (LTP) induction and a prime example of a Hebbian synapse. This pairing, however, has never been actually demonstrated on tetanic stimulation. Optical imaging of neural activity with a voltage-sensitive dye (VSD) is one potentially suitable method for examining this pairing. It is possible with optical recording to examine simultaneously the excitation of postsynaptic neurons at multiple sites. Thus the pairing of spike generation at the soma and dendritic depolarization can be examined with population level optical recording in highly laminar structures such as the hippocampal slice preparation. For example, one can correlate the optical signals obtained from cell layers with the activity of the soma, and, similarly, optical signals from stratum radiatum can be correlated with the activity of the apical dendrite, even though one cannot calibrate the optical signals in terms of actual membrane potential. Using the VSD aminonaphthylethenylpyridinium in rat hippocampal slices, we aimed to examine the pairing. Standard tetanic stimulation (100 Hz, 1 s) that elicited LTP in the field excitatory postsynaptic potential (fEPSP) resulted in a long-lasting depolarizing optical signal (about 2 s) that spread progressively along the known input pathway of CA1. The time course of this long-lasting depolarization was similar to that recorded intracellularly and to that reflected in the fEPSP. The long-lasting depolarization was insensitive to D,L-2-amino-5-phosphonovaleric acid (D,L-APV, 50 microM), but D,L-APV inhibited the induction of LTP; this allowed us to increase the signal-to-noise ratio of the optical signal by averaging several trials. Using this improved optical signal, we confirmed that postsynaptic cells practically "missed" spikes during tetanic stimulation in most parts of CA1, which had been suggested in the intracellular recordings. Intracellular recordings revealed a 23% reduction in input resistance, which might explain the failed spike generation at the soma via shunting. A steep spatial convergence of the depolarization along the transverse axis of area CA1 was observed. In contrast to the response resulting from a standard 100-Hz tetanus, broader activation, and paired depolarization with somatic spikes was observed on theta-burst stimulation. Overall we concluded that postsynaptic spike generation, at least in synchronous form, has less effect on LTP induction with standard tetanic stimulation, while theta-burst tetanic stimulation can elicit pairing of dendritic depolarization and somatic discharge.

Action Potentials↗

Cortical point-spread function and long-range lateral interactions revealed by real-time optical imaging of macaque monkey primary visual cortex.

Processing of retinal images is carried out in the myriad dendritic arborizations of cortical neurons. Such processing involves complex dendritic integration of numerous inputs, and the subsequent output is transmitted to multiple targets by extensive axonal arbors. Thus far, details of this intricate processing remained unexaminable. This report describes the usefulness of real-time optical imaging in the study of population activity and the exploration of cortical dendritic processing. In contrast to single-unit recordings, optical signals primarily measure the changes in transmembrane potential of a population of neuronal elements, including the often elusive subthreshold synaptic potentials that impinge on the extensive arborization of cortical cells. By using small visual stimuli with sharp borders and real-time imaging of cortical responses, we found that shortly after its onset, cortical activity spreads from its retinotopic site of initiation, covering an area at least 10 times larger, in upper cortical layers. The activity spreads at velocities from 100 to 250 microns/msec. Near the V1/V2 border the direct activation is anisotropic and we detected also anisotropic spread; the "space constant" for the spread was approximately 2.7 mm parallel to the border and approximately 1.5 mm along the perpendicular axis. In addition, we found cortical interactions between cortical activities evoked by a small "center stimulus" and by large "surround stimuli" positioned outside the classical receptive field. All of the surround stimuli used suppressed the cortical response to the center stimulus. Under some stimulus conditions iso-orientation suppression was more pronounced than orthogonal-orientation suppression. The orientation dependence of the suppression and its dependency on the size of some specific stimuli indicate that at least part of the center surround inhibitory interaction was of cortical origin. This findings reported here raise the possibility that distributed processing over a very large cortical area plays a major role in the processing of visual information by the primary visual cortex of the primate.

Animals↗

Optical imaging of the spatial distribution of beta-particles emerging from surfaces.

The multiplication in gases of ionization electrons, by the effect of the electric fields between parallel electrodes, leads to the emission of light from the molecules excited in the avalanche process. The optical imaging of this light, with intensifiers, on charge-coupled devices permits the localization, in the gaseous volume, of the entrance points of the beta-particles emitted by radioactive compounds placed close to or at the cathode electrode. Thin slices of anatomical samples labeled with 3H show detailed structures 30 microns in size. Gels carrying 32P or 35S are imaged with accuracies of the order of 0.5 mm (full width at half maximum). In comparison with photographic emulsion, the gain in time for data taking is close to a factor of 100, with the advantage of linearity and wider dynamic range in the intensity measurement and a greatly improved signal-to-noise ratio.

Animals↗

Unwanted optical images with narrow profile intraocular lenses compared with conventional intraocular lenses.

About 35% of intraocular lenses now implanted in the United States are narrow profile lenses, measuring on the average 5.0 mm by 6.0 mm. This study sought to determine whether these lenses cause unwanted optical images with a decrease in the quality of vision. Seventy-five eyes with a 5.0 mm x 6.0 mm intraocular lens (IOL) were compared with 75 eyes with a 6.5 mm IOL. All patients had capsulorhexis, phacoemulsification, and capsular fixation of the IOL. There was no statistically significant difference in unwanted visual images (16% and 20%, respectively), ability to drive at night (79% and 69%), or patient satisfaction. Careful patient selection is, however, important.

Aged↗

Optical imaging of neural activity in auditory cortex induced by intracochlear electrical stimulation.

Little is known about the representation of electrically evoked activity in the auditory cortex. We observed evoked activity in guinea pig auditory cortex evoked by acoustical and electrical stimulation to the cochlea by optical imaging with the aid of a voltage-sensitive dye. Light signals from the cortex were recorded with a 12 x 12 array of photodiodes, and transferred to the spatio-temporal images by every 0.57 ms. The activity by pure tones was shown spatio-temporally through tonotopical organization in the cortex according to the sound frequencies. The tonotopic responses were dynamically changed. When the cochlea was stimulated with single electrical pulses, focal activities were observed in the cortex as spatio-temporal patterns. Activated cortical regions were not sharply localized, but varied with stimulating positions of the cochlea. The curves of response magnitude versus stimulus intensity showed the narrow dynamic range, and that of latency was almost constant. These results were significantly different from those for normal sound stimulation.

Acoustic Stimulation↗

Postsynaptic metabotropic glutamate receptor mGluR1 mediates the late component of signal propagation in the guinea pig piriform cortex: optical imaging study.

Metabotropic glutamate receptors (mGluRs) were previously shown to mediate a postsynaptic late propagation component elicited by layer Ib stimulation in guinea pig piriform cortex slices. In the present study, the effects of some group specific or subtype specific mGluR antagonists on the late propagation component were investigated using an optical imaging method, in order to identify mGluR subtypes mediating it. A selective mGluR1 antagonist (RS)-1-aminoindan-1,5-dicarboxylic acid most effectively suppressed the late component whereas a selective mGluR5 antagonist, selective group II or group III antagonists showed little or no suppressive effect. These results suggest that the late propagation component is mediated by mGluR1.

Animals↗

A new technique for three-dimensional measurements of skin surface contours: evaluation of skin surface contours according to the ageing process using a stereo image optical topometer.

The evaluation of skin surface contours can be carried out by various techniques, including the use of a stylus profilometer, a laserprofilometer and a conventional optical profilometer (COP). But these methods have some drawbacks because their data are basically obtained from two-dimensional algorithms. So a new technique has been developed based on a new concept: a stereo image processing technique which is considered in this paper. Since a pair of stereo images contains depth information, the 'disparity', or the difference between the left and right images, enables the production of three-dimensional coordinates. This study was performed to evaluate the change of skin surface contours according to the ageing process. The stereo image optical topometer (SOT) is a new instrument used for the three-dimensional evaluation of skin surface contours. Thus. five new parameters have to be developed, such as mean surface roughness (S(a)), mean depth of roughness (S(z)), three-dimensional length (S(L)), three-dimensional area (S(A)), and three-dimensional volume (S(V)). S(a), S(L) and S(A) have shown a statistically significant increase in the seventies age group. S(z) has also shown a significant increase in the twenties and over-sixties age groups. The coefficient variation of the height of the skin surface using a COP varies between 14.76 and 6.57, but that using a SOT is between 2.18 and 2.69, according to age variation. In conclusion, the SOT system is a more reliable and useful method for evaluating skin surface contours than the COP system. Among the three-dimensional parameters which were made in this study, S(A), S(L) and S(a) seem to be useful as reliable parameters for evaluating skin surface contours in the ageing process.

Aging↗

Optical imaging of long-term depression in the mouse cerebellar cortex in vivo.

Conjunctive stimulation of climbing fiber and parallel fiber inputs results in long-term depression (LTD) at parallel fiber-Purkinje cell synapses. Although hypothesized to play a major role in cerebellar motor learning, there has been no characterization of the cellular and molecular mechanisms of LTD in the whole animal, let alone its spatial properties, both of which are critical to understanding the role of LTD in cerebellar function. Neutral red optical imaging of the cerebellar cortex in the anesthetized mouse was used to visualize the spatial patterns of activation. Stimulation of the parallel fibers evoked a transverse beam of optical activity, and stimulation of the contralateral inferior olive evoked parasagittal bands. Conjunctive stimulation of parallel fibers and climbing fibers induced a long-term decrease (at least 1 hr) in the optical response to subsequent parallel fiber activation confined to the region of interaction between these two inputs. Activation of climbing fibers alone failed to induce the long-term decrease. Field potential recordings confirmed that the depression is postsynaptic and restricted to the interaction site. The long-term depression in the beam was prevented by a group 1 metabotropic glutamate receptor (mGluR(1)) antagonist and was absent in transgenic mice selectively expressing an inhibitor of protein kinase C (PKC) in Purkinje cells. Conversely, the long-term depression occurred in the mGluR(4) knock-out mouse, consistent with its postsynaptic origin. In addition to providing the first visualization of parallel fiber-Purkinje cell LTD in the cerebellar cortex, this study demonstrates the spatial specificity of LTD and its dependence on mGluR(1) and PKC in vivo.

Animals↗

[The oblique effect revealed by optical imaging in primary visual cortex of cats].

The oblique effect is a ubiquitous visual psychological effect. To explore its underlying neural basis, we quantitatively analyzed the proportion and response amplitude of the cardinal preferred areas and the oblique preferred areas in a fairly large region of the primary visual cortex of cats, using optical imaging based on intrinsic signals. The results show that cardinal preferred areas were larger than oblique preferred areas, with a mean difference of 4.7%. Overall, the responses evoked by cardinal stimuli were generally greater than those by oblique stimuli. The present work provides an explanation for the differences in electrophysiological results reported for this issue, and gives a new insight into the neural basis of the oblique effect.

Animals↗

Synthesis and characterization of glucosamine-bound near-infrared probes for optical imaging.

[structure: see text] Two novel near-infrared (NIR) fluorescent probes have been synthesized by linking a carbocyanine fluorophore and glucosamine through different linkers. These probes demonstrated a high quantum yield, low cytotoxicity, reversible pH-dependent fluorescence in the physiological pH range, and a decreased aggregation tendency in aqueous solutions. In vitro NIR optical imaging studies revealed cellular uptake and strong intracellular NIR fluorescence of these two probes in four breast epithelial cell lines.

Carbocyanines↗

Sub-diffraction-limited optical imaging with a silver superlens.

Recent theory has predicted a superlens that is capable of producing sub-diffraction-limited images. This superlens would allow the recovery of evanescent waves in an image via the excitation of surface plasmons. Using silver as a natural optical superlens, we demonstrated sub-diffraction-limited imaging with 60-nanometer half-pitch resolution, or one-sixth of the illumination wavelength. By proper design of the working wavelength and the thickness of silver that allows access to a broad spectrum of subwavelength features, we also showed that arbitrary nanostructures can be imaged with good fidelity. The optical superlens promises exciting avenues to nanoscale optical imaging and ultrasmall optoelectronic devices.

Journal Article↗

Optical imaging of intrinsic signals as a tool to visualize the functional architecture of adult and developing visual cortex.

One of the most common principles of cortical organization is that neurons with similar response properties are clustered together in space. Thereby the environment is represented in an orderly fashion on the cortical surface in a so-called "cortical map". In primary visual cortex, for instance, neurons with similar orientation preferences are grouped together, forming the orientation preference map. Optical imaging of intrinsic signals allows to investigate the organization of such maps in vivo. Neuronal activity was measured utilizing the fact that the transition from oxy-hemoglobin to hemoglobin in active brain areas can be detected optically by recording changes in light reflectance with a high resolution CCD-camera. When using this technique to look at the exact patterning of orientation preference maps in cat visual cortex a novel principle for the organization of cortical maps was observed: orientation was not organized in parallel bands as had previously been thought but iso-orientation domains were organized radially; orientations from 0 to 180 degrees were laid out in a pinwheel-like fashion around singularities which we termed "orientation-centers". After observing pinwheel patterns in orientation preference maps in adult cat visual cortex it was also investigated how these meticulously arranged maps develop in the cortex of young kittens. Performing chronical recordings in kittens from the age of postnatal day 17 on we were able to observe how orientation maps form already during the third week of life and--under normal conditions--remain largely unchanged thereafter.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Optical imaging of neuronal activity in tissue labeled by retrograde transport of Calcium Green Dextran.

In many neurophysiological studies it is desirable to simultaneously record the activity of a large number of neurons. This is particularly true in the study of vertebrate motor systems that generate rhythmic behaviors, such as the pattern generator for locomotion in vertebrate spinal cord. Optical imaging of neurons labeled with appropriate fluorescent dyes, in which fluorescence is activity-dependent, provides a means to record the activity of many neurons at the same time, while also providing fine spatial resolution of the position and morphology of active neurons. Voltage-sensitive dyes have been explored for this purpose and have the advantage of rapid response to transmembrane voltage changes. However, voltage-sensitive dyes bleach readily, which results in phototoxic damage and limits the time that labeled neurons can be imaged. In addition, the signal-to-noise ratio is typically small, so that averaging of responses is usually required. As an alternative to voltage-sensitive dyes, calcium-sensitive dyes can exhibit large changes in fluorescence. Most neurons contain voltage-sensitive Ca2+ channels, and numerous reports indicate that neuronal activity is accompanied by increased intracellular Ca2+ concentration. In this protocol we describe a method to use retrograde transport of the dextran conjugate of a calcium-sensitive dye (Calcium Green Dextran) to label selectively populations of brain and spinal interneurons in a primitive vertebrate (lamprey), for subsequent video-rate imaging of changes in intracellular fluorescence during neuronal activity. Although described with specific reference to lampreys, the technique has also been applied to embryonic chick spinal cord and larval zebrafish preparations and should be easily adaptable to other systems. The most significant novel feature of the protocol is the use of retrograde axonal transport to selectively fill neurons that have known axonal trajectories. Using lampreys, we have obtained activity-sensitive labeling across longer distances and over a longer transport time (up to 14 mm and 4 days) than has been reported in other species. In addition, retrograde transport allows filling of neurons more deeply within tissue than would be possible with bath application of calcium-sensitive dyes. Furthermore, the dyes are readily taken up by adult tissues, while bath application is usually limited to embryonic and neonatal vertebrate nervous tissues (although the reasons for this limitation are not clear). Attempts to load the AM (acetomethoxy) esters of calcium-sensitive dyes into lamprey spinal cord neurons by bath application have been unsuccessful (McPherson, unpublished observations, and).

Animals↗

Optical imaging in cat area 18: strabismus does not enhance the segregation of ocular dominance domains.

While early-onset strabismus leads to clearly segregated domains of the left and the right eye in cat primary visual cortex (area 17), far less is known about experience-dependent plasticity of ocular dominance in area 18. We therefore used optical imaging of intrinsic signals to analyze the influence of strabismus on cortical maps in cat area 18. Monocular visual stimulation of the left and right eye with moving square wave gratings of four different orientations induced patchy activity maps. Unlike our previous observations in cat area 17, the monocular activity maps in area 18 of strabismic cats were rather similar so that functional ocular dominance domains were not clearly segregated. Imaging of the 17/18 border region confirmed this observation and revealed a sudden change in the segregation of the left and right eye domains across the border. Our results demonstrate that modified visual input can have different consequences for different visual areas: while the decorrelation of activity between the two eyes (as induced by strabismus) clearly enhances the segregation of ocular dominance domains in cat area 17, area 18 does not show this effect although electrophysiological studies have confirmed that the percentage of binocularly driven neurons is as reduced as in area 17.

Animals↗

Optically imaged maps of orientation preference in primary visual cortex of cats and ferrets.

Feature maps in the cerebral cortex constitute orderly representations of response features created within the cortex; an example is the mapping of orientation-selective neurons in visual cortex. We have compared the properties of orientation maps in area 17 of cats and ferrets, obtained by optical imaging of intrinsic signals. Orientation maps in both species contain a quasi-periodic distribution of iso-orientation domains that are organized into a lattice of pinwheels. However, the spatial density of orientation domains and of pinwheels in ferret area 17 is nearly twice that in cat area 17. The ferret map also contains more discontinuities, or fractures, where orientation changes abruptly. The size of orientation domains scales with interdomain spacing, so that the ratio of the two is approximately the same in both species. Consistent with this finding, the orientation tuning width of individual pixels is similar in the two. The magnitude of orientation preference, however, is much lower in ferret compared to cat. The greater incidence of fractures in ferret appears to be due to proportionately greater overlap between domains of different orientations, particularly along fracture lines that link pinwheel centers. We hypothesize that a key determinant of orientation maps, the relationship between orientation domain size and spacing, expresses an anatomical link between sizes of thalamocortical arbors and horizontal intracortical connections in area 17.

Animals↗