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PubMed2004

Thermal AFM: a thermopile case study.

In this work, an atomic force microscope (AFM) with an integrated thermal sensor has been used to obtain the local spatial distribution of temperatures in a micromachined thermopile with submicron resolution. In this communication, we will show how the dimensional, structural and functional characteristics of a thermopile suits well with the requirements for AFM thermal imaging, and how a deeper insight of the thermopile operation can be gained with the aid of these advanced scanning probe-based tools.

Diagnostic Imaging↗
PubMed2005

Non-monotonic changes in performance with eccentricity modeled by multiple eccentricity-dependent limitations.

Eccentricity-dependent resolution losses are sometimes compensated for in psychophysical experiments by magnifying (scaling) stimuli at each eccentricity. The use of either pre-selected scaling factors or unscaled stimuli sometimes leads to non-monotonic changes in performance as a function of eccentricity. We argue that such non-monotonic changes arise when performance is limited by more than one type of constraint at each eccentricity. Building on current methods developed to investigate peripheral perception [e.g., Watson, A. B. (1987). Estimation of local spatial scale. Journal of the Optical Society of America A, 4 (8), 1579-1582; Poirier, F. J. A. M., & Gurnsey, R. (2002). Two eccentricity dependent limitations on subjective contour discrimination. Vision Research, 42, 227-238; Strasburger, H., Rentschler, I., & Harvey Jr., L. O. (1994). Cortical magnification theory fails to predict visual recognition. European Journal of Neuroscience, 6, 1583-1588], we show how measured scaling can deviate from a linear function of eccentricity in a grating acuity task [Thibos, L. N., Still, D. L., & Bradley, A. (1996). Characterization of spatial aliasing and contrast sensitivity in peripheral vision. Vision Research, 36(2), 249-258]. This framework can also explain the central performance drop [Kehrer, L. (1989). Central performance drop on perceptual segregation tasks. Spatial Vision, 4, 45-62] and a case of "reverse scaling" of the integration window in symmetry [Tyler, C. W. (1999). Human symmetry detection exhibits reverse eccentricity scaling. Visual Neuroscience, 16, 919-922]. These cases of non-monotonic performance are shown to be consistent with multiple sources of resolution loss, each of which increases linearly with eccentricity. We conclude that most eccentricity research, including "oddities", can be explained by multiple-scaling theory as extended here, where the receptive field properties of all underlying mechanisms in a task increase in size with eccentricity, but not necessarily at the same rate.

Contrast Sensitivity↗
PubMed2005

Measurement of rate of expansion in the perception of radial motion.

Optic flow generated by rigid surface patches can be decomposed into a small number of elementary motion types. In these experiments, we show that the human visual system can evaluate expansion, one of these motion types, metrically. Moreover, we show that the discrimination of rates of expansion are spatially local. Because the estimation of the focus of expansion is somewhat imprecise, this locality sometimes produces predictable errors in the estimation of rate of expansion. One can make predictions like this with a model adapted from one previously developed for angular-velocity discrimination.

Discrimination, Psychological↗
PubMed2004

Analysis of genes from inner ear developmental-stage cDNA subtraction reveals molecular regionalization of the otic capsule.

Although the gross embryology of inner ear development has been documented for several different vertebrate species at a descriptive level, our understanding of the molecular mechanisms involved remains rudimentary. Therefore, we have used cDNA subtraction and normalization procedures to define genes upregulated in the 13.5dpc mouse inner ear, a developmental stage where inner ear morphogenesis and tissue remodeling is active and differentiation of future hair cells is being initiated. We recovered 33 different genes from this subtraction and using gene-specific primers have confirmed the transcriptional upregulation of 26 of these in the 13.5dpc inner ear. Northern analyses were used to investigate splicing differences between the inner ear and the whole embryo at 13.5dpc. Spatial localization of expression was determined through whole-ear in situ hybridization analysis, and selected genes were analyzed in more detail through in situ hybridization of tissue sections. These data illustrate that the genes isolated in this study are expressed in the developing otic capsule and/or neuroepithelium. Furthermore, the expression patterns also reveal molecular heterogeneity in the developing capsule and indicate that for some genes, the chondrogenic otic capsule is composed of distinct domains of gene expression.

Animals↗
PubMed2005

c-Cbl regulates migration of v-Abl-transformed NIH 3T3 fibroblasts via Rac1.

Cellular events like cell adhesion and migration involve complex rearrangements of the actin cytoskeleton. We have previously shown that the multidomain adaptor protein c-Cbl facilitates actin cytoskeletal reorganizations that result in the adhesion of v-Abl-transformed NIH 3T3 fibroblasts. In this report, we demonstrate that c-Cbl also enhances migration of v-Abl-transformed NIH 3T3 fibroblasts. This effect of c-Cbl depends on its tyrosine phosphorylation, specifically on phosphorylation of its Tyr-731, which is required for binding of PI-3' kinase to c-Cbl. Furthermore, we demonstrate that the effect of c-Cbl on migration of v-Abl-transformed fibroblasts is mediated by active PI-3' kinase and the small GTPase Rac1. Our results also indicate that ubiquitin ligase activity of c-Cbl is required, while spatial localization of c-Cbl to the pseudopodia is not required for the observed effects of c-Cbl on cell migration.

Androstadienes↗
PubMed1990

Differential expression of the murine and rat renin genes in peripheral subcutaneous tissue.

We have previously identified peripheral subcutaneous tissue as a bonafide site of primary renin expression in the mouse fetus by virtue of oncogene mediated tumorigenesis in transgenic mice. In this report we demonstrate that the murine renin genes are differentially expressed in this tissue. Through selective breeding and differential primer extension we demonstrate that Ren-1d and Ren-1c transcripts were several fold more abundant than Ren-2. Renin transcripts were also identified in fetal subcutaneous tissues of Spontaneously Hypertensive (SHR) and Wistar Kyoto (WKY) rats. We conclude from these studies that expression of renin during fetal development may be widespread in rodents with its temporal and spatial localization consistent with a role in fetal development.

Animals↗
PubMed2000

A bimodal pattern of InsP(3)-evoked elementary Ca(2+) signals in pancreatic acinar cells.

InsP(3)-evoked elementary Ca(2+) release events have been postulated to play a role in providing the building blocks of larger Ca(2+) signals. In pancreatic acinar cells, low concentrations of acetylcholine or the injection of low concentrations of InsP(3) elicit a train of spatially localized Ca(2+) spikes. In this study we have quantified these responses and compared the Ca(2+) signals to the elementary events shown in Xenopus oocytes. The results demonstrate, at the same concentrations of InsP(3), Ca(2+) signals consisting of one population of small transient Ca(2+) release events and a second distinct population of larger Ca(2+) spikes. The signal mass amplitudes of both types of events are within the range of amplitudes for the elementary events in Xenopus oocytes. However, the bimodal Ca(2+) distribution of Ca(2+) responses we observe is not consistent with the continuum of event sizes seen in Xenopus. We conclude that the two types of InsP(3)-dependent events in acinar cells are both elementary Ca(2+) signals, which are independent of one another. Our data indicate a complexity to the organization of the Ca(2+) release apparatus in acinar cells, which might result from the presence of multiple InsP(3) receptor isoforms, and is likely to be important in the physiology of these cells.

Animals↗
PubMed2003

Model of intracellular calcium cycling in ventricular myocytes.

We present a mathematical model of calcium cycling that takes into account the spatially localized nature of release events that correspond to experimentally observed calcium sparks. This model naturally incorporates graded release by making the rate at which calcium sparks are recruited proportional to the whole cell L-type calcium current, with the total release of calcium from the sarcoplasmic reticulum (SR) being just the sum of local releases. The dynamics of calcium cycling is studied by pacing the model with a clamped action potential waveform. Experimentally observed calcium alternans are obtained at high pacing rates. The results show that the underlying mechanism for this phenomenon is a steep nonlinear dependence of the calcium released from the SR on the diastolic SR calcium concentration (SR load) and/or the diastolic calcium level in the cytosol, where the dependence on diastolic calcium is due to calcium-induced inactivation of the L-type calcium current. In addition, the results reveal that the calcium dynamics can become chaotic even though the voltage pacing is periodic. We reduce the equations of the model to a two-dimensional discrete map that relates the SR and cytosolic concentrations at one beat and the previous beat. From this map, we obtain a condition for the onset of calcium alternans in terms of the slopes of the release-versus-SR load and release-versus-diastolic-calcium curves. From an analysis of this map, we also obtain an understanding of the origin of chaotic dynamics.

Animals↗
PubMed2003

Mapping mechanical strain of an endogenous cytoskeletal network in living endothelial cells.

A central aspect of cellular mechanochemical signaling is a change of cytoskeletal tension upon the imposition of exogenous forces. Here we report measurements of the spatiotemporal distribution of mechanical strain in the intermediate filament cytoskeleton of endothelial cells computed from the relative displacement of endogenous green fluorescent protein (GFP)-vimentin before and after onset of shear stress. Quantitative image analysis permitted computation of the principal values and orientations of Lagrangian strain from 3-D high-resolution fluorescence intensity distributions that described intermediate filament positions. Spatially localized peaks in intermediate filament strain were repositioned after onset of shear stress. The orientation of principal strain indicated that mechanical stretching was induced across cell boundaries. This novel approach for intracellular strain mapping using an endogenous reporter demonstrates force transfer from the lumenal surface throughout the cell.

Animals↗
PubMed1999

Mucosal mast cell secretion processes imaged using three-photon microscopy of 5-hydroxytryptamine autofluorescence.

The secretion process of the mucosal mast cell line RBL-2H3 was imaged using infrared three photon excitation (3PE) of serotonin (5-hydroxytryptamine, 5-HT) autofluorescence, a measurement previously difficult because of the technical intractability of deep UV optics. Images of prestimulation 5-HT distributions were analyzed in loaded cell populations (those incubated in a 5-HT-rich medium overnight) and in unloaded populations and were found to be strictly quantifiable by comparison with bulk population high-performance liquid chromatography measurements. Antigenically stimulated cells were observed to characteristically ruffle and spread as granular 5-HT disappeared with no detectable granule movement. Individual cells exhibited highly heterogeneous release kinetics, often with quasi-periodic bursts. Neighboring granule disappearances were correlated, indicative of either spatially localized signaling or granule-granule interactions. In one-half of the granule release events, weak residual fluorescence was visible suggestive of leftover 5-HT still bound to the granule matrix. The terminal stages of secretion (>300 s) consisted primarily of unresolved granules and remainder 5-HT leakage from already released granules.

Animals↗
PubMed2002

Rapid changes in expression of glutamate transporters after spinal cord injury.

Glutamate is a major excitatory neurotransmitter in the mammalian CNS. After its release, specific transporter proteins rapidly remove extracellular glutamate from the synaptic cleft. The clearance of excess extracellular glutamate prevents accumulation under normal conditions; however, CNS injury elevates extracellular glutamate concentrations to neurotoxic levels. The purpose of this study was to examine changes in expression and in spatial localization of glial glutamate transporters GLAST (EAAT1) and GLT-1 (EAAT2) and the neuronal glutamate transporter EAAC1 (EAAT3) after spinal cord contusion injury (SCI). The levels of all three transporters significantly increased at the epicenter of injury (T10) and in segments rostral and caudal to the epicenter as determined by Western blot analysis. Quantitative immunohistochemistry demonstrated an increase in GLAST staining in laminae I-V and lamina X both rostral and caudal to the epicenter of injury. Staining for GLT-1 increased significantly in lamina I rostral to the injury site and in the entire gray matter caudal to the injury site. A significant increase in EAAC1 staining was observed in laminae I-IV rostral to the epicenter of injury and throughout the gray matter caudal to the injury site. The results suggest that upregulation of these high affinity transporters occurs rapidly and is important in regulating glutamate homeostasis after SCI.

Amino Acid Transport System X-AG↗
PubMed2002

Expression, regulation and role of the MAGUK protein SAP-97 in human atrial myocardium.

OBJECTIVE: In various cell types, membrane-associated guanylate kinases proteins called MAGUK play a major role in the spatial localization and clustering of ion channels. Here, we studied the expression and role of these anchoring proteins in human right atrial myocardium by means of various molecular, biochemical and physiological methods. METHODS AND RESULTS: SAP-97, PSD-95, Chapsyn and SAP-102 messengers were detected by reverse transcriptase-polymerase chain reaction (RT-PCR) on mRNA extracted from both whole myocardium and isolated myocytes. Western blot revealed that the MAGUK protein SAP-97 and, to a lesser extent, PSD-95, is abundantly expressed in human atrial myocardium, while Chapsyn are almost undetectable. Confocal microscopic visualization of cryosection of atrial myocardium stained with the anti-PSD-95 family antibody showed positive staining at the plasma membrane level and cell extremity. Calpain-I cleaved both SAP-97 and PSD-95 proteins, resulting in an accumulation of short bands, including an 80-kDa band that was also detected in the cytosolic protein fraction. Immunoprecipitation of SAP-97 co-precipitated hKv1.5 channels, and vice versa. Co-expression of cloned SAP-97 and hKv1.5 channels in Chinese hamster ovarian (CHO) cells increased the K(+) current (157.00+/-19.45 pA/pF vs. 344.50+/-58.58 pA/pF at +50 mV). CONCLUSIONS: The protein SAP-97 is abundantly expressed in human atrial myocardium in association with hKv1.5 channels, and probably contributes to regulating the functional expression of the latter.

Adaptor Proteins, Signal Transducing↗
PubMed1999

Controlling cytoskeleton structure by phosphoinositide-protein interactions: phosphoinositide binding protein domains and effects of lipid packing.

Cell movement and resistance to mechanical forces are largely governed by the cytoskeleton, a three-dimensional network of protein filaments that form viscoelastic networks within the cytoplasm. The cytoskeleton underlying the plasma membrane of most cells is rich in actin filaments whose assembly and disassembly are regulated by actin binding proteins that are stimulated or inhibited by signals received and transmitted at the membrane/cytoplasm interface. Inositol phospholipids, or phosphoinositides, are potent regulators of many actin binding proteins, and changes in the phosphorylation of specific phosphoinositide species or in their spatial localization are associated with cytoskeletal remodeling in vitro. This review will focus on recent studies directed at defining the structural features of phosphoinositide binding sites in actin binding proteins and on the influence of the physical state of phosphoinositides on their ability to interact with their target proteins.

Amino Acid Sequence↗
PubMed2002

Dissociating 'what' and 'how' in visual form agnosia: a computational investigation.

Patients with visual form agnosia exhibit a profound impairment in shape perception (what an object is) coupled with intact visuomotor functions (how to act on an object), demonstrating a dissociation between visual perception and action. How can these patients act on objects that they cannot perceive? Although two explanations of this 'what-how' dissociation have been offered, each explanation has shortcomings. A 'pathway information' account of the 'what-how' dissociation is presented in this paper. This account hypothesizes that 'where' and 'how' tasks require less information than 'what' tasks, thereby allowing 'where/how' to remain relatively spared in the face of neurological damage. Simulations with a neural network model test the predictions of the pathway information account. Following damage to an input layer common to the 'what' and 'where/how' pathways, the model performs object identification more poorly than spatial localization. Thus, the model offers a parsimonious explanation of differential 'what-how' performance in visual form agnosia. The simulation results are discussed in terms of their implications for visual form agnosia and other neuropsychological syndromes.

Agnosia↗
PubMed2002

Disturbed visual processing contributes to impaired reading in Alzheimer's disease.

The relationship between visual processing dysfunction and oral reading impairment was investigated in 17 patients with probable or possible Alzheimer's disease (AD). When dementia severity was controlled, a significant relationship was found between single word oral reading impairments and difficulties discriminating words written in different fonts and photographs of objects in different orientations, which are all functions believed to be dependent on the integrity of left ventral temporal-occipital visual association regions. By contrast, there was no significant relationship between reading performance and the score on a test of spatial localization, believed to be more dependent on parietal lobe function. There was also no relationship between reading ability and discrimination of unfamiliar faces, a function thought to engage right inferotemporal lobe structures. In contrast to the significant association between impaired reading and certain visual processes, when dementia severity was controlled, there was no relationship between reading and lexical semantic impairment. These results highlight the contribution of visual processing deficits to impaired oral reading in AD and further suggest that this association may derive from neuropathological changes in areas of the left temporal occipital lobes specialized for high-level visual processing.

Aged↗
PubMed2003

How much does an individual habitat fragment contribute to metapopulation dynamics and persistence?

We derive measures for assessing the value of an individual habitat fragment for the dynamics and persistence of a metapopulation living in a network of many fragments. We demonstrate that the most appropriate measure of fragment value depends on the question asked. Specifically, we analyse four alternative measures: the contribution of a fragment to the metapopulation capacity of the network, to the equilibrium metapopulation size, to the expected time to metapopulation extinction and the long-term contribution of a fragment to colonization events in the network. The latter measure is comparable to density-dependent measures in general matrix population theory, though some differences are introduced by the fact that "density dependence" is spatially localized in the metapopulation context. We show that the value of a fragment depends not only on the properties of the landscape but also on the properties of the species. Most importantly, variation in fragment values between the habitat fragments is greatest in the case of rare species that occur close to the extinction threshold, as these species are likely to be restricted to the most favorable parts of the landscape. We expect that the measures of habitat fragment value described and analysed here have applications in landscape ecology and in conservation biology.

Animals↗
PubMed2000

Spatial color contrast matching: broad-bandpass functions and the flattening effect.

The contrast matching function (CMF) is the reciprocal of test contrast that perceptually matches the contrast of standard pattern, measured as a function of test spatial frequency (SF). Achromatic CMFs usually flatten as the contrast of the standard is raised, and are broader than the achromatic, bandpass, contrast sensitivity function (CSF). This report investigates whether chromatic CMFs have similar characteristics. For this purpose, the red-green color channel was defined using minimum flicker and hue cancellation techniques. Spatially localized (D6), vertical, equiluminant patterns (SFs: 0.063-8 cpd; contrast: 3-80%) were used to measure the CSF and CMF of isoluminant patterns presented with a temporal Gaussian envelope. CMFs were measured using a randomized double-staircase procedure and the two-interval forced choice technique. Two color-normal observers, whose task was to select the interval that had higher color contrast, participated in experiments. Results show that: (a) the color CMFs are lowpass functions of SF at low standard contrasts (3-12.5%), broad-bandpass at intermediate contrasts (6.25-60%), and near-flat at high contrasts (80%); and (b) isoluminant CMFs have higher upper cut-off frequencies than isoluminant CSFs. It is concluded that: (i) color-contrast-constancy (CMF independent of SF) is partly achieved at high contrasts because color CMFs flatten as contrast increases; (ii) the information processing at suprathreshold levels is different from that at the threshold levels; and (iii) the model that explained achromatic CMFs using achromatic threshold mechanisms could not explain chromatic CMFs using chromatic threshold mechanisms.

Adult↗
PubMed2002

Spatial frequency discrimination: a comparison of achromatic and chromatic conditions.

In this study, we have compared foveal SF discriminations for luminance and color-defined stimuli using two different tasks (criteria): in criterion-A, the discrimination is based on spatial (size of the stimuli) and/or spatial frequency; in criterion-B, it is based on apparent motion (contraction/expansion). We used high contrast (75%) spatially localized D6 stimuli and cosine gratings (0.25-9.5 cpd). The SF discrimination was measured by the method of constant stimuli with a two-interval forced-choice procedure. Data show that: (i) for criterion-A, the discrimination thresholds for color stimuli were lower than that for luminance stimuli at low SFs, but similar or higher at higher SFs; for criterion-B, the thresholds to chromatic stimuli were higher than that to achromatic stimuli for all SFs; (ii) SF discrimination was best at inter-stimulus-interval (ISI) of about 200 ms for color stimuli and at ISI of 0 ms for luminance stimuli; (iii) SF discrimination got better with stimulus duration and reached to plateau at 200 ms (or more) for color stimuli and at 67 ms (or more) for luminance stimuli; (iv) SF discrimination threshold (mean Delta(f)=0.19 octaves) is about one-tenth of the full bandwidth (mean=1.96 octaves) of SF tuned mechanisms and is in hyperacuity range; both (discrimination and hyperacuity) can be explained by the relative activities within a population of tuned mechanisms. We conclude that color and luminance SF discrimination thresholds have a different SF dependence. While color appears to perform better than luminance vision at low SFs, this effect is lost or even reversed at high SFs. Data imply that color and form interact, but color and motion are largely segregated (i.e. they weakly interact).

Adult↗