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Glial domains and nerve fiber patterns in the fish retinotectal pathway.

Optic nerve fibers run parallel from the retina as far as the optic tract in fish, then suddenly criss-cross into a new pattern matching the tectal map. This change coincides with a unique demarcation between two astroglial territories in the retinotectal pathway, located where the optic chiasm occurs in other vertebrates, which we defined using antibodies directed against intermediate filaments (IF). We found that astroglia in optic nerve territory express an Mr 56,000 IF polypeptide, band 3, which we identify as the fish equivalent of vimentin in mammals. These astrocytic cells lack glial fibrillary acidic protein (GFAP; cf. Dahl and Bignami, 1973). Conversely, glia in brain territory, that is, in the optic tract and elsewhere in the CNS, lack the fish vimentin, but express GFAP. By electron microscopy, we obtained evidence that new retinal axons extend swiftly through the growing optic nerve, where they are tightly shepherded into a narrow track by newly differentiating glial cells, positive for the fish vimentin. In the GFAP-positive glial territory of the optic tract, by contrast, growing axons are slowed down and probably branch. We suggest that this allows them to fasciculate accurately with older fibers and thereby propagate a tectotopic pattern established by pioneer axons in the embryo.

Animals↗

Disconnected optic axons persist in the visual pathway during regeneration of the retino-tectal projection in the frog.

In this study, we crushed one optic nerve in the frog Litoria (Hyla) moorei and at intervals thereafter anterogradely labelled optic axons with horseradish peroxidase (HRP). For one series, HRP was applied between the eye and the crush site and in a second series between the crush site and the chiasm. A tectal projection of regenerating axons was seen in both series but, in addition, up to 12 weeks post-crush, the second series displayed an additional projection. Its appearance matched that of the disconnected, but persisting, optic axon terminals which are found after enucleation or optic nerve ligation. We conclude that, in the frog, many disconnected optic axons persist throughout the period of optic nerve regeneration and of restoration of an orderly retino-tectal map.

Animals↗

Light dosimetry for intraperitoneal photodynamic therapy in a murine xenograft model of human epithelial ovarian carcinoma.

Few studies have been published to date measuring spatially resolved fluence rates in complex tissue geometries. Here the light distributions of three different intraperitoneal light delivery geometries in a murine ovarian cancer model were investigated to assess their influence on the tumorcidal efficacy of photodynamic therapy (PDT). In vivo fluence rate measurements in the peritoneal cavities of mice, with the light intensity being mapped in three transverse planes, were performed using fiber-optic detectors. Three different source fiber designs and placements were tested for their ability to provide uniform irradiation of the peritoneal cavity. The biological response to a PDT protocol comprising three separate treatments administered at 72 h intervals, each consisting of a 0.25 mg kg-1 intraperitoneal injection of benzoporphyrin derivative-mono acid ring A followed 90 min later by delivery of 15 J of 690 nm light, was measured. The tissue response was evaluated by measuring the number of remaining visible lesions and the total residual tumor mass. Fluence rate measurements showed large variations in the fluence rate distribution for similar intended treatments. The most uniform and reproducible illumination was achieved using two 18 mm long cylindrical emitting optical fibers. The biological response was comparable to that produced when a flat-cleaved end optical fiber is used to illuminate the four quadrants of the abdomen sequentially. While a good reproducibility in tumor induction in this animal model exists, no correlation was found between the fluence rate distribution measured in one group of animals and the biological response in a separate group of similarly treated animals. Due to the large intra-animal variability in fluence rate distribution, representative fluence rate mapping in complex tissue geometries is of limited value when applied to an individual PDT treatment. Thus, surveillance of the fluence rate during individual treatments will be required for acceptable PDT dosimetry. To improve the versatility of this particular animal model for PDT research, a large number of extended sources are required to increase uniformity of the illumination in order to reduce unwanted cytotoxic side effects resulting from foci of high fluence rates. In this way, subsequent increase of the total energy delivered to the tumor may be possible.

Animals↗

Map plasticity in somatosensory cortex.

Sensory maps in neocortex are adaptively altered to reflect recent experience and learning. In somatosensory cortex, distinct patterns of sensory use or disuse elicit multiple, functionally distinct forms of map plasticity. Diverse approaches-genetics, synaptic and in vivo physiology, optical imaging, and ultrastructural analysis-suggest a distributed model in which plasticity occurs at multiple sites in the cortical circuit with multiple cellular/synaptic mechanisms and multiple likely learning rules for plasticity. This view contrasts with the classical model in which the map plasticity reflects a single Hebbian process acting at a small set of cortical synapses.

Animals↗

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.

Animals↗

Functional reorganization of visual cortex maps after ischemic lesions is accompanied by changes in expression of cytoskeletal proteins and NMDA and GABA(A) receptor subunits.

Reorganization of cortical representations after focal visual cortex lesions has been documented. It has been suggested that functional reorganization may rely on cellular mechanisms involving modifications in the excitatory/inhibitory neurotransmission balance and on morphological changes of neurons peripheral to the lesion. We explored functional reorganization of cortical retinotopic maps after a focal ischemic lesion in primary visual cortex of kittens using optical imaging of intrinsic signals. After 1, 2, and 5 weeks postlesion (wPL), we addressed whether functional reorganization correlated in time with changes in the expression of MAP-2, GAP-43, GFAP, GABA(A) receptor subunit alpha1 (GABA(A)alpha1), subunit 1 of the NMDA receptor (NMDAR1), and in neurotransmitter levels at the border of the lesion. Our results show that: (1) retinotopic maps reorganize with time after an ischemic lesion; (2) MAP-2 levels increase gradually from 1wPL to 5wPL; (3) MAP-2 upregulation is associated with an increase in dendritic-like structures surrounding the lesion and a decrease in GFAP-positive cells; (4) GAP-43 levels reach the highest point at 2wPL; (5) NMDAR1 and glutamate contents increase in parallel from 1wPL to 5wPL; (6) GABA(A)alpha1 levels increase from 1wPL to 2wPL but do not change after this time point; and (7) GABA contents remain low from 1wPL to 5wPL. This is a comprehensive study showing for the first time that functional reorganization correlates in time with dendritic sprouting and with changes in the excitatory/inhibitory neurotransmission systems previously proposed to participate in cortical remodeling and suggests mechanisms by which plasticity of cortical representations may occur.

Amino Acids↗

[The data processing of neurophysiological examination and the diagnostic support system].

Neurophysiological examinations include EEG, EMG, Evoked Potential (EP) and ENG. In this paper, EEG and EP data processing, and the diagnostic support system using these findings were discussed. The FFT and AR model for analysis of frequency, and the pattern recognition for spike or spindle detection are the technical methods for application to EEG data processing of the first order. Amplitude or phase mapping techniques, as a second order data processing, were not only applied to the diagnosis of neurological disorders, but also used for detection of electrical equivalent dipole source localization which was inversely reconstructed in the cerebral cortex from the distribution of EEG potentials on the scalp. The averaging technique was used for detection of small evoked potentials (EP) such as SEP or ABR. The following items were included in the diagnostic support system applied to data processing. 1) Topographic analysis in the brain mapping of SEP to median nerve stimulation was applied to the identification of the central sulcus during a neurosurgical procedure to maintain the QOL of patient. 2) The usefulness of EEG automatic reporting system with Japanese sentences and brain topographic mapping was described. 3) Digital EEG in a data filing system using magneto optical disks was used for data analysis and diagnostic support system after EEG examination.

Algorithms↗

Genetic and molecular characterization of the optomotor-blind gene locus in Drosophila melanogaster.

The Drosophila gene optomotor-blind (omb) is involved in the development of a set of giant neurons in the optic lobes and possibly other structures in the imaginal brain. Adult flies have discrete defects in optomotor behavior. The gene has previously been mapped in chromomeres 4C5-6, together with three other genes, bifid, Quadroon and lacqueredgls. We have localized the gene in a genomic walk of 340 kb of DNA. By mapping seven chromosome breakpoints with omb phenotype we determined its minimum size to about 80 kb. From this region more than 20 RNAs of different size and temporal expression pattern are transcribed. Three of them (T3, T7 and T7') stem from primary transcripts of 40-80 kb in length. In its distal part the omb gene overlaps in at least 19 kb with four other complementation units, bifid, l(1)bifid, Quadroon and lacqueredgls. The three nonlethals affect the external appearance of the fly and seem to be unrelated to brain development.

Animals↗

From diffusion tractography to quantitative white matter tract measures: a reproducibility study.

The aim of this study is to propose methods for assessing the reproducibility of diffusion tractography algorithms in future clinical studies and to show their application to the tractography algorithm developed in our unit, fast marching tractography (FMT). FMT estimates anatomical connectivity between brain regions using the information provided by diffusion tensor imaging. Three major white-matter pathways were investigated in 11 normal subjects--anterior callosal fibers, optic radiations, and pyramidal tracts. FMT was used to generate maps of connectivity metric, and regions of voxels with highest connectivity metric to an anatomically defined starting point were identified for each tract under investigation. The reproducibilities of tract-"normalized" volume (NV) and fractional anisotropy (FA) measurements were assessed over such regions. The values of tract volumes are consistent with the postmortem data. Coefficients of variation (CVs) for FA and NV ranged from 1.7 to 7.1% and from 2.2 to 18.6%, respectively. CVs were lowest in the anterior callosal fibers (range: 1.7- 7.8%), followed by the optic radiations (range: 1.2-18.6%) and pyramidal tracts (range: 2.6-15.5%), suggesting that fiber organization plays a role in determining the level of FMT reproducibility. In conclusion, these findings underline the importance of assessing the reliability of diffusion tractography before investigating white matter pathology.

Adult↗

Temporal and spatial effects of Sonic hedgehog signaling in chick eye morphogenesis.

Proper dorsal--ventral pattern formation of the optic cup is essential for vertebrate eye morphogenesis and retinotectal topographic mapping. Previous studies have suggested that midline tissue-derived Sonic hedgehog (Shh) molecules play critical roles in establishing the bilateral eye fields and in determining the proximal--distal axis of the eye primordium. Here, we have examined the temporal requirements for Shh during the optic vesicle to optic cup transition and after early optic cup formation in chick embryos. Both misexpressing Shh by virus and blocking Shh activity by antibodies resulted in disruption of ventral ocular tissues. Decreasing endogenous Shh signals unexpectedly revealed a sharp morphological boundary subdividing dorsal and ventral portions of the optic cup. In addition, Shh signals differentially influenced expression patterns of genes involved in ocular tissue specification (Pax6, Pax2, and Otx2) and dorsal--ventral patterning (cVax) within the ventral but not dorsal optic cup. Ectopic Shh suppressed expression of Bone Morphogenetic Protein 4 (BMP4) in the dorsal retina, whereas reducing endogenous Sonic hedgehog activity resulted in a ventral expansion of BMP4 territory. These results demonstrate that temporal requirements for Shh signals persist after the formation of the optic cup and suggest that the early vertebrate optic primordium may be subdivided into dorsal and ventral compartments. We propose a model in which ventrally derived Shh signals and dorsally restricted BMP4 signals act antagonistically to regulate the growth and specification of the optic primordium.

Animals↗

Retinal projections in the freshwater butterfly fish, Pantodon buchholzi (Osteoglossoidei). II. Differential projections of the dorsal and ventral hemiretinas.

Pantodon buchholzi, the freshwater butterfly fish, is a member of the Osteoglossomorpha, the most primitive of the four major teleost radiations. The projections of fibers originating in the dorsal and ventral hemiretinas in Pantodon, as determined with autoradiography, are reported here. Fibers originating in the ventral hemiretina reach their targets through the axial, medial and dorsal optic tracts. Fibers that originate in the dorsal hemiretina reach their points of termination by way of the axial, medial and ventral optic tracts. Projections of the various tracts to preoptic, thalamic, tubercular, pretectal and tectal regions, as described in the previous study of total retinal projections, were verified. The retinal projections to the preoptic, thalamic and tubercular nuclei do not map topographically. Ventral hemiretinal fibers are mapped, however, onto the dorsal part of the nucleus pretectalis superficialis pars parvocellularis, the rostral part of the dorsal accessory optic nucleus, the entire nucleus pretectalis periventricularis pars ventralis and the dorsomedial portion of the optic tectum. Ventral hemiretinal fibers also supply most if not all the retinal innervation to the central pretectal nucleus. In contrast, dorsal hemiretinal fibers are mapped onto the ventral part of nucleus pretectalis superficialis pars parvocellularis, the entire dorsal accessory optic nucleus and the ventrolateral portion of the optic tectum. The dorsal and ventral hemiretinal projections to the tectum about at a cytoarchitectonically recognizable point, indicating that no discontinuity is present in the retinal connectivity with the tectum. The pars parvocellularis of nucleus pretectalis superficialis is a simple, unfolded, and nonlaminar structure in Pantodon. This structure contrasts markedly with the more complex, folded structure of the nucleus in the majority of other examined teleosts. The orientation of the projections from the dorsal and ventral hemiretinas onto this nucleus in Pantodon is congruent with that seen in other fishes only after a schematic unfolding of the nucleus in these fishes.

Animals↗

Near-field mapping of surface polariton fields.

Using the general approach to image formation in collection near-field optical microscopy, I derive the symmetry relations for the amplitude coupling coefficients in the case of a weakly guiding single-mode fibre terminated with a probe tip possessing axial symmetry. It is shown that, for the symmetrical detection configuration, six elements of the coupling matrix can be expressed by using only three independent coupling coefficients. The obtained relations are further applied to describe near-field mapping of surface plasmon polariton (SPP) fields. I demonstrate that, for the symmetrical detection configuration, the near-field optical image reflects the intensity distribution of the SPP field components parallel to the surface plane, even though the strong perpendicular component is also being detected. This conclusion is supported with numerical simulations that elucidate the influence of symmetry of the fibre probe on the resulting near-field optical image. The near-field optical images simulated for scattering systems typical for SPP microoptics and localization are presented. It is found that the presence of asymmetry in the detection configuration increases the contribution of the perpendicular field component and results in the images approaching the corresponding SPP intensity distributions.

Journal Article↗

Diffusion-tensor imaging in septo-optic dysplasia.

One of the morphological correlates of septo-optic dysplasia is hypoplasia of the optic nerves. As of now, it remains unknown, in how far this disorder also affects the organization of the optic radiation. Using diffusion-tensor imaging (DTI), the non-invasive evaluation of large fiber tracts including the optic radiation has become possible. We have compared DTI-data from a patient suffering from septo-optic dysplasia with those of a group of eleven healthy control subjects. The anisotropy showed statistically significant reduction in the patient with septo-optic dysplasia within the visual fiber tracts and an unordered eigenvector map. A comparison of the anisotropy in the pyramidal tract showed no significant difference. Since the patient was congenitally blind, it remains unclear whether the findings are the results of the underlying disorder or occur in all congenitally blind patients. One might presume, that, in order for the optic radiation to fully develop, an afferent input to the lateral geniculate body is necessarry.

Adult↗

The organization of regenerating axons in the Xenopus optic nerve.

After regeneration of the optic nerve in Xenopus, with restoration of an orderly visuotectal map, HRP was applied to a small region of retina, or tectum, and the distribution of labelled fibres in the nerve investigated. As they passed the site of the crush, fibres became grossly disordered and remained so until they got to the chiasma.

Animals↗

Chronically mountable goggles for persistent exposure to single orientation.

To examine the effect of experience on the developmental plasticity of functional maps in the visual cortex, we need to establish a method for a stable visual experience manipulation under the freely moving condition. For this purpose, we fabricated goggles that are chronically mounted stably on the animal's head, but easy to replace according to the animal's growth. Here we report the design of the goggles and the method of mounting them on the head of animals. By this method, combined with the intrinsic signal optical imaging technique, we were able to observe a rapid and robust reorganization of orientation maps.

Aging↗

Projections of the nucleus of the basal optic root in pigeons (Columba livia) revealed with biotinylated dextran amine.

The nucleus of the basal optic root (nBOR) of the accessory optic system is known to be involved in the analysis of the visual consequences of self-motion. Previous studies have shown that the nBOR in pigeons projects bilaterally to the vestibulocerebellum, the inferior olive, the interstitial nucleus of Cajal, and the oculomotor complex and projects unilaterally to the ipsilateral pretectal nucleus lentiformis mesencephali and the contralateral nBOR. By using the anterograde tracer biotinylated dextran amine, we confirmed these projections and found (previously unreported) projections to the nucleus Darkshewitsch, the nucleus ruber, the mesencephalic reticular formation, and the area ventralis of Tsai as well as ipsilateral projections to the central gray, the pontine nuclei, the cerebellar nuclei, the vestibular nuclei, the processus cerebellovestibularis, and the dorsolateral thalamus. In addition to previous studies, which showed a projection to the dorsomedial subdivision of the contralateral oculomotor complex, we found terminal labelling in the ventral and dorsolateral subdivisions. Individual fibers were reconstructed from serial sections, and collaterals to various nuclei were demonstrated. For example, collaterals of fibers projecting to the vestibulocerebellum terminated in the vestibular or cerebellar nuclei; collaterals of fibers to the inferior olive terminated in the pontine nuclei; many individual neurons projected to the interstitial nucleus of Cajal, the nucleus Darkshewitsch, and the central gray and also projected to the nucleus ruber and the mesencephalic reticular formation; collaterals of fibers to the contralateral nucleus of the basal optic root terminated in the mesencephalic reticular formation and/or the area ventralis of Tsai; neurons projecting to the nucleus lentiformis mesencephali also terminated in the dorsolateral thalamus. The consequences of these data for understanding the visual control of eye movements, neck movements, posture, locomotion, and visual perception are discussed.

Animals↗

Use of intrinsic optical signals to monitor physiological changes in brain tissue slices.

Optical imaging techniques have the potential to bring a combination of high spatial and temporal resolution to studies of brain function. Many optical techniques require the addition of a dye or fluorescent marker to the tissue, and such methods have proven extremely valuable. It is also known that the intrinsic optical properties of neural tissue are affected by certain physiological changes and that these intrinsic optical signals can provide information not available by other means. Most authors attribute the intrinsic optical change to alterations in cell volume and concomitant change in the concentration of the cytosol. In this article we review the literature on intrinsic optical signals, covering both the mechanisms of the optical change and its use in various branches of neurophysiology. We also discuss technical aspects of the technique as used with hippocampal slices, including illumination methods, cameras, experimental methods, and data collection and analysis procedures. Finally we present data from investigations in which we used intrinsic optical signals in hippocampal slices to study the extent of spread of synaptic activation, propagation of spreading depression, extent and severity of the response to hypoxia, and tissue response to osmotic challenges. We conclude that (1) at least two processes generate intrinsic optical signals in hippocampal slices, one of which causes light scattering to change inversely with cell volume and is related to dilution of the cytoplasm, while the other, opposite in sign, may be due to mitochondrial swelling; and (2) the intrinsic optical signal can be a useful tool for spatial mapping of relatively slow events, but is not suitable for study of fast physiological processes.

Animals↗

Synaptogenesis in the first optic neuropile of the fly's visual system.

The developmental transformation of the chief afferent class of fly photoreceptor synapse has been examined from serial electron micrographs of animals fixed at 74, 81, 94 and 100% pupal development (100% pupal development being defined by the time of normal adult eclosion). Animals were selected both by their age and the conformity of their eye coloration to standards for each stage. Two animals were analysed from each stage, one in greater detail than the other. For the first, the exact coordinates of the cartridges (the synaptic columns of the first optic neuropile) from which the analyses were made were mapped and selected to be within the same region of the eye field at all stages. From all animals a portion of one or two cartridges was analysed from series of up to 100 sections and the synapse populations (greater than 80) were analysed for their fine structure and postsynaptic composition. Adult synapses are confirmed as tetrads, with two of the four postsynaptic elements invariably from two monopolar interneurons L1 and L2, one from each. The two others are usually from alpha processes of the same amacrine cell. Synapses appear during the last half of pupal development, with no obvious asynchrony of ultrastructural maturation and in parallel with those of at least one of the other synaptic classes present (which were otherwise not studied). Many adult features of synaptic ultrastructure emerge late, only by 94% pupal development. These include adult numbers of synaptic vesicles, the complete form of the presynaptic ribbon with platform and the postsynaptic cisternae of L1/L2. Prior to 94% the synapses are smaller with postsynaptic elements having a less regular geometry and with postsynaptic densities which are subsequently lost (alpha processes) or replaced by cisternae (L1/L2). At the presynaptic sites of the younger animals (74%, 81%) dyads and triads of postsynaptic elements coexist with tetrads, those of older animals having, on average, more postsynaptic processes per synapse. It is suggested that individual synapses assemble piecemeal, element by element.

Afferent Pathways↗