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Shifter circuits: a computational strategy for dynamic aspects of visual processing.

We propose a general strategy for dynamic control of information flow between arrays of neurons at different levels of the visual pathway, starting in the lateral geniculate nucleus and the geniculorecipient layers of cortical area V1. This strategy can be used for resolving computational problems arising in the domains of stereopsis, directed visual attention, and the perception of moving images. In each of these situations, some means of dynamically controlling how retinal outputs map onto higher-level targets is desirable--in order to achieve binocular fusion, to allow shifts of the focus of attention, and to prevent blurring of moving images. The proposed solution involves what we term "shifter circuits," which allow for dynamic shifts in the relative alignment of input and output arrays without loss of local spatial relationships. The shifts are produced in increments along a succession of relay stages that are linked by diverging excitatory inputs. The direction of shift is controlled at each stage by inhibitory neurons that selectively suppress appropriate sets of ascending inputs. The shifter hypothesis is consistent with available anatomical and physiological evidence on the organization of the primate visual pathway, and it offers a sensible explanation for a variety of otherwise puzzling facts, such as the plethora of cells in the geniculorecipient layers of V1.

Animals↗

Enhanced c-myc gene expression during forelimb regenerative outgrowth in the young Xenopus laevis.

Analysis of the expression of the c-myc protooncogene has been carried out in the forelimb regenerate of the Xenopus laevis froglet. Northern blot hybridization analysis revealed the presence of a 2.5-kilobase c-myc transcript in the regenerate forelimb at a level at least 7-fold more than the one found in nonregenerating forelimbs or stumps of regenerating forelimbs. In situ hybridization analyses confirmed the relative abundance of c-myc RNA in the regenerate forelimb and provided evidence of spatial localization of high levels of c-myc RNA in specific cell layers. The deepest layers of the wound epithelium of epidermal origin showed a strong signal, whereas virtually no c-myc RNA was detected in the outermost layers. Labeling was also observed in mesenchymal cells of the blastema where it was relatively evenly distributed. This pattern of c-myc RNA in the regenerate might indicate that the expression of c-myc plays a role in the regulation of the continued proliferation of specific cells of the regenerate, whereas repression of this gene in the epidermis correlates with terminal differentiation of keratinocytes.

Amputation, Surgical↗

Casimir light: field pressure.

The electromagnetic field is assigned a self-consistent role in which abrupt slowing of the collapse produces radiation and the pressure of the radiation produces abrupt slowing. A simple expression is introduced for the photon spectrum. Conditions for light emission are proposed that imply a high degree of spatial localization. Some numerical checks are satisfied. A study of the mechanical equations of motion suggests an explanation of the very short time scale in terms of oppositely directed field pressures and the speed of light.

Journal Article↗

Scalp electrical potentials reflect regional cerebral blood flow responses during processing of written words.

Functional brain imaging studies with positron emission tomography (PET) have identified blood flow changes in widely separated areas of the brain during the performance of word-related tasks. In the present study, we have utilized event-related electrical potentials (ERPs) to investigate the temporal relationships among cortical areas previously identified by PET to be differentially activated when performing a task involving generating the uses of visually presented nouns versus reading aloud. ERPs showed strong task-related differences over left and middle inferior frontal and left parietotemporal regions. Frontal and left parietotemporal channels revealed these differences around 200 and 700 msec, respectively, after word presentation. These results provide the time course for parts of the anatomical circuit involved in generating the meaning of a word. Our results also demonstrate how combining the spatial localization of PET with the temporal resolution of ERPs greatly enhances the capacity to understand the mechanisms involved in human cognition.

Adult↗

The neural development and organization of letter recognition: evidence from functional neuroimaging, computational modeling, and behavioral studies.

Although much of the brain's functional organization is genetically predetermined, it appears that some noninnate functions can come to depend on dedicated and segregated neural tissue. In this paper, we describe a series of experiments that have investigated the neural development and organization of one such noninnate function: letter recognition. Functional neuroimaging demonstrates that letter and digit recognition depend on different neural substrates in some literate adults. How could the processing of two stimulus categories that are distinguished solely by cultural conventions become segregated in the brain? One possibility is that correlation-based learning in the brain leads to a spatial organization in cortex that reflects the temporal and spatial clustering of letters with letters in the environment. Simulations confirm that environmental co-occurrence does indeed lead to spatial localization in a neural network that uses correlation-based learning. Furthermore, behavioral studies confirm one critical prediction of this co-occurrence hypothesis, namely, that subjects exposed to a visual environment in which letters and digits occur together rather than separately (postal workers who process letters and digits together in Canadian postal codes) do indeed show less behavioral evidence for segregated letter and digit processing.

Adult↗

Neurotrophins regulate agrin-induced postsynaptic differentiation.

The precise orchestration of synaptic differentiation is critical for efficient information exchange in the nervous system. The nerve-muscle synapse forms in response to agrin, which is secreted from the motor nerve terminal and induces the clustering of acetylcholine receptors (AChRs) and other elements of the postsynaptic apparatus on the subjacent muscle cell surface. In view of the highly restricted spatial localization and the plasticity of neuromuscular junctions, it seems likely that synapse formation and maintenance are regulated by additional, as-yet-unidentified factors. Here, we tested whether neurotrophins modulate the agrin-induced differentiation of postsynaptic specializations. We show that both brain-derived neurotrophic factor (BDNF) and neurotrophin-4 (NT-4) inhibit agrin-induced AChR clustering on cultured myotubes. Nerve growth factor and NT-3 are without effect. Muscle cells express full-length TrkB, the cognate receptor for BDNF and NT-4. Direct activation of this receptor by anti-TrkB antibodies mimicked the BDNF/NT-4 inhibition of agrin-induced AChR clustering. This BDNF/NT-4 inhibition is likely to be an intrinsic mechanism for regulating AChR clustering, because neutralization of endogenous TrkB ligands resulted in elevated levels of AChR clustering even in the absence of added agrin. Finally, high concentrations of agrin can occlude the BDNF/NT-4 inhibition of AChR clustering. These results indicate that an interplay between agrin and neurotrophins can regulate the formation of postsynaptic specializations. They also suggest a mechanism for the suppression of postsynaptic specializations at nonjunctional regions.

Agrin↗

Targeted cytoplasmic irradiation with alpha particles induces mutations in mammalian cells.

Ever since x-rays were shown to induce mutation in Drosophila more than 70 years ago, prevailing dogma considered the genotoxic effects of ionizing radiation, such as mutations and carcinogenesis, as being due mostly to direct damage to the nucleus. Although there was indication that alpha particle traversal through cellular cytoplasm was innocuous, the full impact remained unknown. The availability of the microbeam at the Radiological Research Accelerator Facility of Columbia University made it possible to target and irradiate the cytoplasm of individual cells in a highly localized spatial region. By using dual fluorochrome dyes (Hoechst and Nile Red) to locate nucleus and cellular cytoplasm, respectively, thereby avoiding inadvertent traversal of nuclei, we show here that cytoplasmic irradiation is mutagenic at the CD59 (S1) locus of human-hamster hybrid (AL) cells, while inflicting minimal cytotoxicity. The principal class of mutations induced are similar to those of spontaneous origin and are entirely different from those of nuclear irradiation. Furthermore, experiments with radical scavenger and inhibitor of intracellular glutathione indicated that the mutagenicity of cytoplasmic irradiation depends on generation of reactive oxygen species. These findings suggest that cytoplasm is an important target for genotoxic effects of ionizing radiation, particularly radon, the second leading cause of lung cancer in the United States. In addition, cytoplasmic traversal by alpha particles may be more dangerous than nuclear traversal, because the mutagenicity is accomplished by little or no killing of the target cells.

Alpha Particles↗

Quantitative assessment of Hox complex expression in the indirect development of the polychaete annelid Chaetopterus sp.

A prediction from the set-aside theory of bilaterian origins is that pattern formation processes such as those controlled by the Hox cluster genes are required specifically for adult body plan formation. This prediction can be tested in animals that use maximal indirect development, in which the embryonic formation of the larva and the postembryonic formation of the adult body plan are temporally and spatially distinct. To this end, we quantitatively measured the amount of transcripts for five Hox genes in embryos of a lophotrochozoan, the polychaete annelid Chaetopterus sp. The polychaete Hox complex is shown not to be expressed during embryogenesis, but transcripts of all measured Hox complex genes are detected at significant levels during the initial stages of adult body plan formation. Temporal colinearity in the sequence of their activation is observed, so that activation follows the 3'-5' arrangement of the genes. Moreover, Hox gene expression is spatially localized to the region of teloblastic set-aside cells of the later-stage embryos. This study shows that an indirectly developing lophotrochozoan shares with an indirectly developing deuterostome, the sea urchin, a common mode of Hox complex utilization: construction of the larva, whether a trochophore or dipleurula, does not involve Hox cluster expression, but in both forms the complex is expressed in the set-aside cells from which the adult body plan derives.

Animals↗

The Lin12-notch repeats of pregnancy-associated plasma protein-A bind calcium and determine its proteolytic specificity.

The Lin12-Notch repeat (LNR) module of about 35 residues is a hallmark of the Notch receptor family. Three copies, arranged in tandem, are invariably present in the extracellular portion of the Notch receptors. Although their function is unknown, genetic and biochemical data indicate that the LNR modules participate in the regulation of ligand-induced proteolytic cleavage of the Notch receptor, a prerequisite to intramembrane cleavage and Notch signaling. Outside the Notch receptor family, the LNR module is present only in the metalloproteinase pregnancy-associated plasma protein-A (PAPP-A) and its homologue PAPP-A2, which also contain three copies. Curiously, LNR modules 1 and 2 are present within the proteolytic domain of PAPP-A/A2, but LNR3 is separated from LNR2 by more than 1000 amino acids. The growth factor antagonists insulin-like growth factor-binding protein (IGFBP)-4 and -5 are both substrates of PAPP-A. We provide here evidence that the PAPP-A LNR modules function together to determine the proteolytic specificity of PAPP-A. Analysis of C-terminally truncated PAPP-A mutants followed by the analysis of LNR deletion mutants demonstrated that each of the three PAPP-A LNR modules is strictly required for proteolytic activity against IGFBP-4 but not for proteolytic activity against IGFBP-5. Individual substitution of conserved LNR residues predicted to participate in calcium coordination caused elimination (D341A, D356A, D389A, D1484A, D1499A, and D1502A) or a significant reduction (D359A and E392A) of IGFBP-4 proteolysis, whereas IGFBP-5 proteolysis was unaffected. The activity of the latter mutants against IGFBP-4 could be partially rescued by calcium, and the addition of the calcium-binding protein calbindin D9k to wild-type PAPP-A eliminated activity against IGFBP-4 but not against IGFBP-5, demonstrating that the PAPP-A LNR modules bind calcium ions. We propose a model in which LNR3 is spatially localized in proximity to LNR1 and -2, forming a single functional unit.

Amino Acid Sequence↗

Actin activation of myosin heavy chain kinase A in Dictyostelium: a biochemical mechanism for the spatial regulation of myosin II filament disassembly.

Studies in Dictyostelium discoideum have established that the cycle of myosin II bipolar filament assembly and disassembly controls the temporal and spatial localization of myosin II during critical cellular processes, such as cytokinesis and cell locomotion. Myosin heavy chain kinase A (MHCK A) is a key enzyme regulating myosin II filament disassembly through myosin heavy chain phosphorylation in Dictyostelium. Under various cellular conditions, MHCK A is recruited to actin-rich cortical sites and is preferentially enriched at sites of pseudopod formation, and thus MHCK A is proposed to play a role in regulating localized disassembly of myosin II filaments in the cell. MHCK A possesses an aminoterminal coiled-coil domain that participates in the oligomerization, cellular localization, and actin binding activities of the kinase. In the current study, we show that the interaction between the coiled-coil domain of MHCK A and filamentous actin leads to an approximately 40-fold increase in the initial rate of kinase catalytic activity. Actin-mediated activation of MHCK A involves increased rates of kinase autophosphorylation and requires the presence of the coiled-coil domain. Structure-function analyses revealed that the coiled-coil domain alone binds to actin filaments (apparent K(D) = 0.9 microm) and thus mediates the direct interaction with F-actin required for MHCK A activation. Collectively, these results indicate that MHCK A recruitment to actin-rich sites could lead to localized activation of the kinase via direct interaction with actin filaments, and thus this mode of kinase regulation may represent an important mechanism by which the cell achieves localized disassembly of myosin II filaments required for specific changes in cell shape.

Actins↗

Development of methodology for the three-dimensional modelling of the metabolic capacity of the rat nasal cavity using glutathione S-transferase M1 as an example.

A variety of chemicals induce site-specific lesions in the rodent nasal cavity. In order to explore the reasons for this site-selectivity, methodology for (a) creation of a 3-dimensional (3D) model of a rat nasal cavity, and (b) mapping of semiquantitative data onto the model has been developed. The head of a rat was fixed, decalcified, step-sectioned (every 100 microm) and stained with hematoxylin and eosin. Digital images of the sections were optically captured, and a KS400 image analysis system (Imaging Associates, Thame, Oxford, UK), attached to a standard personal computer, was used to align adjacent images and reconstruct the series in 3D. The final model was anatomically correct, and could be rotated in any plane and manipulated to display individual internal structures. The spatial localization of a glutathione S-transferase (rGSTM1, previously known as GST 3-3) within this model was investigated using immunohistochemistry. Step sections (every 400 microm) were stained, analyzed by imaging densitometry, and the results for the stained regions within the nasal cavity divided into 4 grades representing high to low expression of rGSTM1. The data was mapped onto the 3D model and showed that the highest expression of this enzyme was in the central regions of the nasal cavity at the transition between respiratory and olfactory epithelia. This methodology will allow investigation of the relationship between the in situ localization of bioactivating and detoxifying enzyme systems and the site-specificity of nasal lesions.

Animals↗

Differential effects of prime-probe duration on positive and negative location priming: Evidence for opponent facilitatory and inhibitory influences in priming tasks.

A series of four spatial localization experiments is reported that examined the effects of display duration and presentation mode on positive and negative priming using an attended-repetition and an ignored-repetition paradigm, respectively. Experiment 1 showed larger positive priming with response-dependent than with 150 ms display durations while negative priming remained unaffected. Experiments 2-4 were performed to further elucidate the effects of prime-probe durations. Data suggest largely independent effects of prime and probe duration on priming effects. Manipulation of prime duration affected facilitation due to repetition of the prime distractor location as well as inhibitory effects associated with ignored repetition. Furthermore, anticipated probe duration modulated the effectiveness of inhibition of return. Findings are discussed within a framework proposing two major components of priming effects--a stimulus-driven or automatic component, and a strategic component related to the participant's expectations towards the probe.

Adult↗

Training the adult amblyopic eye with "perceptual learning" after vision loss in the non-amblyopic eye.

We recently reported acuity development in the amblyopic eye of a 60-year-old patient after loss of vision in her non-amblyopic eye. Here, we focus on the training that we implemented, based on new insights from psychophysical procedures aiming at functional visual improvement of adults ("perceptual learning"). We alternately used the following procedures: grating acuity (Teller-Cards); contrast sensitivity (Vistech-Charts); two spatial localization tests (vertical alignment, pointing); and labyrinth patterns for a eye-hand coordination exercise. One month without intervention was followed by six months of training and two blocks of pleoptic treatment. Clinical parameters were assessed monthly. Besides acuity gain, we observed enhanced grating resolution and contrast sensitivity, decreased alignment distortions, pointing shifts, mainly after pleoptics, and more efficient labyrinth tracing. A questionnaire reflected the patient's perception of the changes. These data confirm the plasticity of the adult amblyopic system, be it spontaneous due to the loss of the non-amblyopic eye or caused by the intervention or both. Further experience is necessary to isolate the role of the intervention. Our results also underline the limitation of adult plasticity, emphasizing the importance of early diagnosis and treatment of amblyopia.

Amblyopia↗

Superfund soil cleanup: developing the Piazza Road remedial design.

A statistical approach was used to develop the remediation and soil sampling strategy at the Piazza Road dioxin site (an EPA Superfund site in Missouri). The source of the dioxin was contaminated waste oil that was used as a dust suppressant. This site was used as a test case to determine if a "surgical" remedial design could be developed. This new approach was compared to the historical approach taken by U.S. EPA Region 7 in Missouri. A pilot study provided information on the local spatial pattern of dioxin, so that sampling designs could be evaluated and costs of remediation could be estimated for different remedial strategies. Sampling designs were evaluated by a Monte Carlo approach, where the laboratory analytical error and sampling error due to spatial variation of dioxin were modeled based on the results of the pilot study and the historical site data. The optimal cleanup unit size was determined to be 14x14 ft, or 1/24th of the historical cleanup unit size. A companion paper describes the performance achieved and the dollar 5,900,000 cost savings that resulted from applying the improved design. This case study clearly shows the value of environmental data, where the use and quality of the data are established as a part of planning the site remediation.

Costs and Cost Analysis↗

Pontine and lateral reticular projections to the c1 zone in lobulus simplex and paramedian lobule of the rat cerebellar cortex.

Spatial localization and axonal branching in mossy fiber projections to two rostrocaudally-separated regions of the 'forelimb' c1 zone in lobulus simplex and paramedian lobule were studied in rats using a retrograde double-labelling tracer technique. In four animals the two cortical regions were localized electrophysiologically and each was micro-injected with tracer material, yielding a total of eight different cases. Single- and double-labelled cell bodies were plotted in the basal pontine nucleus (BPN), nucleus reticularis tegmenti pontis (NRTP), and the lateral reticular nucleus (LRN). As a control, cells labelled in the contralateral inferior olive were also counted. The parts of the c1 zone in lobulus simplex and the paramedian lobule were found to receive mossy fiber inputs from similar regions of BPN, NRTP and LRN. Double-labelled cells were not found in NRTP but were present in BPN and LRN (on average 6% and 25% of the smaller single-labelled population, respectively). The incidence of double-labelled cells in the olive and LRN was positively correlated, but no relation was found between olive and BPN, suggesting a zonal organization within the mossy fiber projections from LRN, but not from the pons. In quantitative terms, the c1 zone in lobulus simplex received a greater density of mossy fiber projections from BPN, NRTP and LRN than the c1 zone in the paramedian lobule. This suggests that the two parts of the same cerebellar cortical zone differ, at least partially, in regard to their inputs from three major sources of mossy fibers. This is consistent with the modular hypothesis and could enable a higher degree of parallel processing and integration of information within different parts of the same zone.

Animals↗

Cellular and subcellular distribution of a cAMP-regulated prestalk protein and prespore protein in Dictyostelium discoideum: a study on the ontogeny of prestalk and prespore cells.

We have analyzed a developmentally and spatially regulated prestalk-specific gene and a prespore-specific gene from Dictyostelium. The prestalk gene, pst-cathepsin, encodes a protein highly homologous to the lysosomal cysteine proteinases cathepsin H and cathepsin B. The prespore gene encodes a protein with some homology to the anti-bacterial toxin crambin and has been designated beejin. Using the lambda gtll system, we have made polyclonal antibodies directed against a portion of the protein encoded by pst-cathepsin and other antibodies directed against the beejin protein. Both antibodies stain single bands on Western blots. By immunofluorescence and Western blots, pst-cathepsin is not present in vegetative cells or developing cells during the first approximately 10 h of development. It then appears with a punctate distribution in a subset of developing cells. Beejin is detected only after approximately 15 h of development, also in a subset of cells. Pst-cathepsin is distributed in the anterior approximately 1/10 of migrating slugs and on the peripheral posterior surfaces of slugs. Beejin is distributed in the posterior region of slugs. Expression of both pst-cathepsin and beejin can be induced in subsets of isolated cultured cells by a combination of conditioned medium and extracellular cAMP in agreement with the regulation of the mRNAs encoding these proteins. We have used the antibodies as markers for cell type to examine the ontogeny and the spatial distribution of prestalk and prespore cells throughout multicellular development. Our findings suggest that prestalk cell differentiation is independent of position within the aggregate and that the spatial localization of prestalk cells within the multicellular aggregate arises from sorting of the prestalk cells after their induction. We have also found a class of cell in developing aggregates that contains neither the prestalk nor the prespore markers.

Antibodies, Monoclonal↗

Actin-binding proteins from Drosophila embryos: a complex network of interacting proteins detected by F-actin affinity chromatography.

By using F-actin affinity chromatography columns to select proteins solely by their ability to bind to actin filaments, we have identified and partially purified greater than 40 proteins from early Drosophila embryos. These proteins represent approximately 0.5% of the total protein present in soluble cell extracts, and 2 mg are obtained by chromatography of an extract from 10 g of embryos. As judged by immunofluorescence of fixed embryos, 90% of the proteins that we have detected in F-actin column eluates are actin-associated in vivo (12 of 13 proteins tested). The distributions of antigens observed suggest that groups of these proteins cooperate in generating unique actin structures at different places in the cell. These structures change as cells progress through the cell cycle and as they undergo the specializations that accompany development. The variety of different spatial localizations that we have observed in a small subset of the total actin-binding proteins suggests that the actin cytoskeleton is a very complex network of interacting proteins.

Actins↗

Shootin1: A protein involved in the organization of an asymmetric signal for neuronal polarization.

Neurons have the remarkable ability to polarize even in symmetrical in vitro environments. Although recent studies have shown that asymmetric intracellular signals can induce neuronal polarization, it remains unclear how these polarized signals are organized without asymmetric cues. We describe a novel protein, named shootin1, that became up-regulated during polarization of hippocampal neurons and began fluctuating accumulation among multiple neurites. Eventually, shootin1 accumulated asymmetrically in a single neurite, which led to axon induction for polarization. Disturbing the asymmetric organization of shootin1 by excess shootin1 disrupted polarization, whereas repressing shootin1 expression inhibited polarization. Overexpression and RNA interference data suggest that shootin1 is required for spatially localized phosphoinositide-3-kinase activity. Shootin1 was transported anterogradely to the growth cones and diffused back to the soma; inhibiting this transport prevented its asymmetric accumulation in neurons. We propose that shootin1 is involved in the generation of internal asymmetric signals required for neuronal polarization.

Amino Acid Sequence↗