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Results for “Spatial heterogeneity”

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At least 883 records · Page 49Linked to original sources

PubMed2006

Use of confocal laser as light source reveals stomata-autonomous function.

In most terrestrial plants, stomata open during the day to maximize the update of CO(2) for photosynthesis, but they close at night to minimize water loss. Blue light, among several environmental factors, controls this process. Stomata response to diverse stimuli seems to be dictated by the behaviour of neighbour stomata creating leaf areas of coordinated response. Here individual stomata of Arabidopsis leaves were illuminated with a short blue-light pulse by focusing a confocal argon laser. Beautifully, the illuminated stomata open their pores, whereas their dark-adapted neighbours unexpectedly experience no change. This induction of individual stomata opening by low fluence rates of blue light was disrupted in the phototropin1 phototropin2 (phot1 phot2) double mutant, which exhibits insensitivity of stomatal movements in blue-illuminated epidermal strips. The irradiation of all epidermal cells making direct contact with a given stoma in both wild type and phot1 phot2 plants does not trigger its movement. These results unravel the stoma autonomous function in the blue light response and illuminate the implication of PHOT1 and/or PHOT2 in such response. The micro spatial heterogeneity that solar blue light suffers in partially shaded leaves under natural conditions highlights the physiological significance of the autonomous stomatal behaviour.

Arabidopsis↗

PubMed2002

Intracellular concentration measurements in adherent cells: a comparison of import efficiencies of cell-permeable peptides.

A protocol was developed for performing intracellular concentration measurements in flat adherent tissue culture cells by fluorescence correlation microscopy (FCM). Determination of the number of molecules in the confocal detection volume had to account for background fluorescence caused by molecules adsorbed to the surface of the measurement chamber. Such a background signal leads to a decrease in the amplitude of the autocorrelation function, and thereby to the calculation of an erroneously high number of molecules. Because of the spatial heterogeneity of the background intensity, a method was devised by which its contribution to the total fluorescence could be determined directly from each individual autocorrelation measurement. This method was applied to a comparison of the import efficiencies of different cell-permeable peptides at nanomolar concentrations. The Antennapedia homeodomain-derived peptide penetratin was imported about three times as efficient as the basic fibroblast growth factor-derived MTS peptide. Both peptides equilibrated between cytoplasm and nucleus. A relatively high mobility of these molecules inside the cells indicated that they may be rapidly degraded by cytosolic proteases. Based on these results, it will be possible to determine intracellular concentrations of inhibitors linked to import peptides directly by FCM at nanomolar concentrations and to optimise such constructs for proteolytic stability.

Carrier Proteins↗

PubMed2006

The influence of aridity and fire on holocene prairie communities in the eastern Prairie Peninsula.

The role of climate and fire in the development, maintenance, and species composition of prairie in the eastern axis of the tallgrass Prairie Peninsula intrigued early North American ecologists. However, evaluation of the long-standing hypotheses about the region's environmental history has been hampered by the scarcity of paleorecords. We conducted multiproxy analyses on early and middle Holocene sediments from two Illinois, USA, lakes to assess long-term climatic, vegetational, and fire variability in the region. Sediment mineral composition, carbonate delta18O, ostracode assemblages, and diatom assemblages were integrated to infer fluctuations in moisture availability. Pollen and charcoal delta13C were used to reconstruct vegetation composition, and charcoal influx was used to reconstruct fire. Results indicate that fire-sensitive trees (e.g., Ulmus, Ostrya, Fraxinus, and Acer saccharum) declined and prairie taxa expanded with increased aridity from 10,000 yr BP to 8500 yr BP. Between approximately 8500 yr BP and approximately 6200 yr BP, aridity declined, and prairie coexisted with fire-sensitive and fire-tolerant (e.g., Quercus and Carya) trees. After approximately 6200 yr BP, prairie taxa became dominant, although aridity was not more severe than it was around 8500 yr BP. Along with aridity, fire appears to have played an important role in the establishment and maintenance of prairie communities in the eastern Prairie Peninsula, consistent with the speculations of the early ecologists. Comparison of our data with results from elsewhere in the North American midcontinent indicates that spatial heterogeneity is a characteristic feature of climatic and vegetational variations on millennial time scales.

Animals↗

PubMed2006

Prey risk allocation in a grazing ecosystem.

Understanding the behaviorally mediated indirect effects of predators in ecosystems requires knowledge of predator-prey behavioral interactions. In predator-ungulate-plant systems, empirical research quantifying how predators affect ungulate group sizes and distribution, in the context of other influential variables, is particularly needed. The risk allocation hypothesis proposes that prey behavioral responses to predation risk depend on background frequencies of exposure to risk, and it can be used to make predictions about predator-ungulate-plant interactions. We determined non-predation variables that affect elk (Cervus elaphus) group sizes and distribution on a winter range in the Greater Yellowstone Ecosystem (GYE) using logistic and log-linear regression on surveys of 513 1-km2 areas conducted over two years. Employing model selection techniques, we evaluated risk allocation and other a priori hypotheses of elk group size and distributional responses to wolf (Canis lupus) predation risk while accounting for influential non-wolf-predation variables. We found little evidence that wolves affect elk group sizes, which were strongly influenced by habitat type and hunting by humans. Following predictions from the risk allocation hypothesis, wolves likely created a more dynamic elk distribution in areas that they frequently hunted, as elk tended to move following wolf encounters in those areas. This response should dilute elk foraging pressure on plant communities in areas where they are frequently hunted by wolves. We predict that this should decrease the spatial heterogeneity of elk impacts on grasslands in areas that wolves frequently hunt. We also predict that this should decrease browsing pressure on heavily browsed woody plant stands in certain areas, which is supported by recent research in the GYE.

Animals↗

PubMed2006

Sensitivity of ecological models to their climate drivers: statistical ensembles for forcing.

Global and regional numerical models for terrestrial ecosystem dynamics require fine spatial resolution and temporally complete historical climate fields as input variables. However, because climate observations are unevenly spaced and have incomplete records, such fields need to be estimated. In addition, uncertainty in these fields associated with their estimation are rarely assessed. Ecological models are usually driven with a geostatistical model's mean estimate (kriging) of these fields without accounting for this uncertainty, much less evaluating such errors in terms of their propagation in ecological simulations. We introduce a Bayesian statistical framework to model climate observations to create spatially uniform and temporally complete fields, taking into account correlation in time and space, spatial heterogeneity, lack of normality, and uncertainty about all these factors. A key benefit of the Bayesian model is that it generates uncertainty measures for the generated fields. To demonstrate this method, we reconstruct historical monthly precipitation fields (a driver for ecological models) on a fine resolution grid for a climatically heterogeneous region in the western United States. The main goal of this work is to evaluate the sensitivity of ecological models to the uncertainty associated with prediction of their climate drivers. To assess their numerical sensitivity to predicted input variables, we generate a set of ecological model simulations run using an ensemble of different versions of the reconstructed fields. We construct such an ensemble by sampling from the posterior predictive distribution of the climate field. We demonstrate that the estimated prediction error of the climate field can be very high. We evaluate the importance of such errors in ecological model experiments using an ensemble of historical precipitation time series in simulations of grassland biogeochemical dynamics with an ecological numerical model, Century. We show how uncertainty in predicted precipitation fields is propagated into ecological model results and that this propagation had different modes. Depending on output variable, the response of model dynamics to uncertainty in inputs ranged from uncertainty in outputs that matched that of inputs to those that were muted or that were biased, as well as uncertainty that was persistent in time after input errors dropped.

Animals↗

PubMed2006

The impact of exploiting grazers (Scaridae) on the dynamics of Caribbean coral reefs.

Coral reefs provide a number of ecosystem services including coastal defense from storms, the generation of building materials, and fisheries. It is increasingly clear that the management of reef resources requires an ecosystem approach in which extractive activities are weighed against the needs of the ecosystem and its functions rather than solely those of the fishery. Here, I use a spatially explicit simulation model of a Caribbean coral reef to examine the ecosystem requirements for grazing which is primarily conducted by parrotfishes (Scaridae). The model allows the impact of fishing grazers to be assessed in the wider context of other ecosystem processes including coral-algal competition, hurricanes, and mass extinction of the herbivorous urchin Diadema antillarum. Using a new analytical model of scarid grazing, it is estimated that parrotfishes can only maintain between 10% and 30% of a structurally complex forereef in a grazed state. Predictions from this grazing model were then incorporated into a broader simulation model of the ecosystem. Simulations predict that scarid grazing is unable to maintain high levels of coral cover (> or = 30%) when severe hurricanes occur on a decadal basis, such as occurs in parts of the northern Caribbean. However, reefs can withstand such intense disturbance when grazing is undertaken by both scarids and the urchin Diadema. Scarid grazing is predicted to allow recovery from hurricanes when their incidence falls to 20 years or less (e.g., most of Central and South America). Sensitivity analyses revealed that scarid grazing had the most acute impact on model behavior, and depletion led to the emergence of a stable, algal-dominated community state. Under conditions of heavy grazer depletion, coral cover was predicted to decline rapidly from an initial level of 30% to less than 1% within 40 years, even when hurricane frequency was low at 60 years. Depleted grazers caused a population bottleneck in juvenile corals in which algal overgrowth caused elevated levels of postsettlement mortality and resulted in a bimodal distribution of coral sizes. Several new hypotheses were generated including a region-wide change in the spatial heterogeneity of coral reefs following extinction of Diadema. The management of parrotfishes on Caribbean reefs is usually approached implicitly through no-take marine reserves. The model predicts that depletion of grazers in nonreserve areas can severely limit coral accretion. Other studies have shown that low coral accretion can reduce the structural complexity and therefore quality of the reef habitat for many organisms. A speculative yet rational inference from the model is that failure to manage scarid populations outside reserves will have a profoundly negative impact on the functioning of the reserve system and status of non-reserve reefs.

Animals↗

PubMed2005

Polychlorinated biphenyls and toxaphene in preferred prey fish of coastal southeastern U.S. bottlenose dolphins (Tursiops truncatus).

Legacy organochlorine (OC) contaminants continue to pose a potential risk to ecological and human health in coastal aquatic ecosystems of the southeastern United States. Polychlorinated biphenyls (PCBs) and toxaphene (TOX) were analyzed by gas chromatography with electron capture detection and negative chemical ionization mass spectrometry in 77 composites of four inshore fish species commonly preyed upon by bottlenose dolphins (Tursiops truncatus) from estuaries near Savannah, Georgia (SAV), Brunswick, Georgia (BRN), and Jacksonville, Florida (JAX), USA. Whereas seasonal and species-specific differences were minimal, differences among mean total PCB concentrations (sigmaPCBs) by estuary (42.0 +/- 48.3, 1.59 +/- 1.24, and 0.281 +/- 0.075 microg/g lipid for BRN, JAX, and SAV, respectively) were highly significant. This estuary-specific trend also held true for mean total toxaphene concentrations (sigmaTOX): 49 +/- 100 (BRN), 1.2 +/- 0.52 (JAX), and 0.40 +/- 0.19 microg/g lipid (SAV). Congener profiles of PCBs also were found to be significantly different among estuaries, with BRN and (to a lesser extent) JAX samples enriched with highly chlorinated homologs associated with Aroclor 1268, a legacy OC linked to a historical point source in Brunswick. The observed spatial heterogeneity in OC concentrations and PCB congener profiles suggests that contaminated fish from Brunswick pose the greatest risk to ecological and human health via biomagnification and seafood consumption; highly chlorinated PCBs (and possibly toxaphene) are transported in a southerly, alongshore direction; and the uniqueness of Aroclor 1268 underscores its utility as a signature proxy in future regional ecotoxicological studies.

Animals↗

PubMedDate not supplied

Spatial variability of soil carbon in forested and cultivated sites: implications for change detection.

The potential to sequester atmospheric carbon in agricultural and forest soils to offset greenhouse gas emissions has generated interest in measuring changes in soil carbon resulting from changes in land management. However, inherent spatial variability of soil carbon limits the precision of measurement of changes in soil carbon and hence, the ability to detect changes. We analyzed variability of soil carbon by intensively sampling sites under different land management as a step toward developing efficient soil sampling designs. Sites were tilled cropland and a mixed deciduous forest in Tennessee, and old-growth and second-growth coniferous forest in western Washington, USA. Six soil cores within each of three microplots were taken as an initial sample and an additional six cores were taken to simulate resampling. Soil C variability was greater in Washington than in Tennessee, and greater in less disturbed than in more disturbed sites. Using this protocol, our data suggest that differences on the order of 2.0 Mg C ha(-1) could be detected by collection and analysis of cores from at least five (tilled) or two (forest) microplots in Tennessee. More spatial variability in the forested sites in Washington increased the minimum detectable difference, but these systems, consisting of low C content sandy soil with irregularly distributed pockets of organic C in buried logs, are likely to rank among the most spatially heterogeneous of systems. Our results clearly indicate that consistent intramicroplot differences at all sites will enable detection of much more modest changes if the same microplots are resampled.

Agriculture↗

PubMed2003

Transformations for cDNA microarray data.

Two channel microarray data often contain systematic variations that can be minimized by data transformation prior to further analysis. The most commonly observed effects are revealed by viewing scatter plots of the logarithm of the ratio by the average logarithmic intensity of the two color channels (RI plots). In this paper we present a general model for signal intensity data with multiple error sources. We demonstrate how these sources of error influence the shape of an RI plot. We then compare some currently available transformation strategies in terms of their mechanism and performance on both simulated and real microarray data. A linlog transformation is proposed to stabilize the variance of the log ratios. We also propose a regional smoothing method to remove variation in log ratios due to spatial heterogeneity on the microarray surface. The discussed transformations represent an important initial step in microarray data analysis for both ratio-based and ANOVA methods.

Journal Article↗

PubMed1995

Visualization of endothelial clefts and nuclei in living microvessels with combined reflectance and fluorescence confocal microscopy.

OBJECTIVE: The cellular basis for spatial heterogeneity along postcapillary venules in their response to inflammatory mediators is not understood. To study permeability regulatory processes on an individual cell basis in intact microvessels, we developed methods to delineate individual endothelial cells within living, single perfused microvessels. METHODS: Individual postcapillary microvessels in the mesenteries of frogs and hamsters were perfused with chloride-free Ringer's solutions containing AgNO3 (0.1 g per 100 ml) for 5--10 s. Vessels were immediately flushed with Ringer's solution containing serum albumin and some vessels subsequently perfused with the fluorescent nucleic acid stain YO-PRO-1 (1 microM) for 10-15 min. Vessels were imaged in situ using laser scanning confocal microscopy. A reflectance image from silver precipitate in the endothelial clefts and a fluorescence image of the nuclei were simultaneously recorded. Stacks of confocal images were merged and used to reconstruct the three-dimensional orientation of endothelial cells. Hydraulic conductivity of some frog mesenteric venular vessels, measured before and after perfusion with AgNO3, was used to assess the integrity of the vessels. RESULTS: The endothelial cell clefts were delineated with a very fine, readily imaged precipitate of silver. Hydraulic conductivity measured after AgNO3 perfusion in frog mesenteric venular microvessels was not significantly different from control. Vessels showed a slight reduction in reactivity to inflammatory stimuli. The three-dimensional pattern of endothelial cells in the living vessels was imaged. CONCLUSIONS: Combining confocal microscopy with silver staining in single perfused microvessels was shown to be an effective means to delineate the three-dimensional pattern of endothelial cells forming microvessel walls. It enables further study of the vascular structure and function relationships at the individual cell level in intact microvessels.

Animals↗

PubMed1997

Blood flow in the cerebral capillary network: a review emphasizing observations with intravital microscopy.

Capillary perfusion in the brain is characterized by an essentially continuous flow of erythrocytes and plasma in almost all capillaries. Rapid fluctuations and spatial heterogeneity or red blood cell (RBC) velocity (0.5-1.8 mm/s) within the capillary network are present. In addition, low-frequency (4-8 cpm) synchronous oscillations in RBC velocity in the capillary network emerge when perfusion to cerebral tissue is challenged. Despite the tortuous, three-dimensional architecture of microvessels, functional intercapillary anastomoses are absent. At rest, red cells travel through the capillary network in 100-300 ms along 150- to 500-micron-long paths. Physiological challenges elicit sizable changes in RBC velocity with a minor role for capillary recruitment, change in capillary diameter, or flow shunting. During acute hypoxia, RBC velocity increases in all capillaries; the corresponding response to hypereapnia is more complex and involves redistribution of capillary flow toward more homogeneous perfusion. The response of capillary flow to decreased perfusion pressure reflects autoregulation of cerebral blood flow but also involves intranetwork redistribution of RBC flow between two populations of capillaries, postulated as thoroughfare channels and exchange capillaries. Flow reserve may be provided by the thoroughfare channels and may help maintain flow velocity and capillary exchange and protect the microcirculation from perfusion failure. Isovolemic hemodilution increases RBC velocity three- to fourfold and increases RBC flux to a moderate degree with a relatively small decrease in capillary hematocrit, under normal and compromised arterial blood supply. In cerebral ischemia, leukocyte adhesion is enhanced and appears reversible when the ischemia is moderate but may be progressive when the injury is severe. The observed flow behavior suggests the presence of a physiological regulatory mechanism of cerebral capillary flow that may involve communication among various microvascular and parenchymal cells and utilize locally acting endothelial and parenchymal mediators such as endothelium-derived relaxing factor or nitric oxide.

Animals↗

PubMed2005

Medulloblastoma: mouse models and novel targeted therapies based on the Sonic hedgehog pathway.

Understanding molecular pathways, signaling cascades, and genetic alterations activated during tumorigenesis is essential for the development of targeted cancer treatments. In children, tumors of the central nervous system are thought to arise from progenitor cells that show considerable temporal and spatial heterogeneity in a developmental environment that is different from that of the adult. Investigating the molecular basis of pediatric tumors is critical because it is likely to generate novel treatments. Animal models have brought many important advances in this field. In this review the authors discuss the mouse models based on the Sonic hedgehog pathway, which have provided a better knowledge of the genetic and molecular alterations of medulloblastoma.

Animals↗

PubMed2026

Metagenomic insights into antibiotic resistance genes and virulence factors in sediments of river Yamuna.

Riverine sediments serve as critical reservoirs of microbial diversity and functional genes, reflecting both natural ecological processes and anthropogenic impacts. In the present study, we employed a shotgun metagenomic approach to investigate microbial community composition, antimicrobial resistance (AMR) genes, and virulence factors in sediments collected from three environmentally distinct locations of the Yamuna River near Agra, India, representing BSA, TGY, and YEA. The sediment DNA was subjected to high-throughput Illumina sequencing, followed by quality control, assembly, and open reading frame prediction. Taxonomic classification and diversity analyses were performed using MEGAN6 and R-based statistical tools, while AMR genes were identified from predicted metagenomic proteins using the Resistance Gene Identifier (RGI) against the CARD database, with high-confidence perfect and strict hits retained; ARGs were interpreted independently of species-level host assignment. Virulence factors were assessed through presence-absence profiling of functionally relevant gene categories. The results revealed pronounced spatial heterogeneity in microbial communities, with increasing taxonomic diversity, functional complexity, and evenness from BSA to TGY and YEA. TGY and YEA composite samples showed greater observed representation of high-confidence AMR gene predictions spanning multiple drug classes and resistance mechanisms, alongside a diverse repertoire of virulence-associated genes linked to motility, adhesion, and secretion systems. In contrast, the BSA site harbored a comparatively simpler resistome and virulome. Overall, this study highlights Yamuna River sediments as important reservoirs of resistance and virulence determinants and underscores the need for long-term genomic surveillance to inform risk assessment, pollution control, and sustainable river management strategies.

AMR↗

PubMed2026

Deep learning-based cross-attention fusion of multimodal MRI for survival prediction and risk stratification in IDH-wildtype glioblastoma: a multicenter study.

BACKGROUND: Glioblastoma (GBM) exhibits profound molecular and spatial heterogeneity, complicating prognostic evaluations. While multiparametric MRI provides crucial multidimensional biological information, conventional end-to-end deep learning integration strategies, such as early or late fusion, often fail to capture complex nonlinear cross-modal interactions. We aimed to systematically evaluate a cross-attention fusion (CAF) architecture for GBM survival prediction and quantify its incremental prognostic value relative to existing clinical tools. METHODS: In this multicenter retrospective study, 386 adults with IDH-wildtype, WHO grade 4 GBM were assembled from an institutional cohort (n = 226), the Chinese Glioma Genome Atlas (CGGA, n = 62), and The Cancer Genome Atlas (TCGA, n = 98). Using a unified 3D ResNet-18 backbone, we compared single-modality models, early fusion, late fusion, and CAF on preoperative T1-weighted, contrast-enhanced T1-weighted (T1CE), and T2-weighted MRI, and integrated the resulting deep learning risk score with routine clinical variables through multivariable Cox regression. Performance was assessed using Harrell's C-index, time-dependent AUC, and decision curve analysis. RESULTS: CAF showed numerically higher, more consistent C-index trends than early fusion, late fusion, and single-modality models (pooled C-index 0.629, 95% CI 0.594-0.664), although pairwise differences in time-dependent AUC were not statistically significant. Integrating clinical variables raised the pooled C-index to 0.691 (95% CI 0.660-0.721) in the treatment-era model, with comparable performance across the three cohorts (Local 0.688; CGGA 0.716; TCGA 0.689); a pre-treatment configuration excluding adjuvant therapy yielded a pooled C-index of 0.642. Under leave-one-cohort-out external validation, the combined model retained significant risk stratification in all held-out cohorts (C-index 0.63-0.71; all log-rank P < 0.01), albeit with attenuated discrimination. The deep learning risk score remained independent after multivariable adjustment (HR 1.41 per SD, 95% CI 1.26-1.57; P < 0.001). Kaplan-Meier analysis confirmed significant high- versus low-risk separation in all cohorts, and decision curve analysis showed greater net benefit than clinical-only and deep-learning-only models. CONCLUSION: The CAF-derived risk score offers prognostic information complementary to routine clinical variables, representing a promising noninvasive tool for individualized risk stratification when molecular profiling is incomplete or unavailable; these findings warrant prospective external validation before clinical use.

cross-attention fusion↗

PubMed1998

Epidemiology of endemic Oropouche virus transmission in upper Amazonian Peru.

A cross-sectional serosurvey of a rural community near Iquitos, Peru was conducted to determine Oropouche (ORO) virus antibody prevalence and risk factors for human infection. Venous blood samples, and demographic, social, and risk factor data were obtained from people age five years of age and older who lived in the village of Santa Clara on the Nanay River, a tributary of the Amazon River. Sera were tested for ORO viral antibody by an ELISA. The specificity of viral antibody reactivity was determined by a standard plaque-reduction neutralization test. Interview data were analyzed by univariate and multiple logistic regression to determine which variables were statistically associated with previous ORO viral infection, as indicated by the presence of IgG antibody. Final models were evaluated based on log-likelihood and Wald chi-square. Clustering of seropositive residents within houses was analyzed by the method of Walter. Among 1,227 persons sampled, 33.7% (n=414) were positive for ORO viral IgG antibody. Overall, antibody prevalence was similar for males (33.9%) and females (33.6%), and increased significantly with age for both sexes to include more than half of persons more than 25 years of age. The length of residence in the village was positively associated with serologic status; persons who had moved to the village within the past 15 years were less likely to be seropositive than life-long residents of the same age. Antibody prevalence among immigrants who had lived in Santa Clara more than 15 years was similar to that in life-long residents. The activity most predictive of previous ORO viral infection was travel to forest communities and travel to Iquitos. No evidence of spatial heterogeneity in ORO virus antibody distribution was observed. Results suggested that endemic transmission of ORO virus in this region has been ongoing during many decades, and that people are at considerable risk of infection.

Adolescent↗

PubMed1999

New techniques for clinical assessment of the peripheral microcirculation.

Current methods for clinical investigation of the cutaneous microcirculation in patients are based mainly on laser Doppler and capillary microscopy. The use of laser Doppler gives a semi-quantitative index of superficial tissue perfusion. The most recent devices are capable of analysing both the volumetric and velocimetric components. New instruments use two different frequencies to compare tissue perfusion at different depths beneath the skin surface. The combination of a laser probe and a small automate produces a 2-dimensional image, allowing the evaluation of spatial heterogeneity in tissue perfusion, an important pathophysiological concept in vascular diseases. Capillaroscopy has recently been improved by the emergence of the flexible videomicroscope, allowing easy exploration of not only the classical site of the nail-fold but also of the body skin surface. The use of this method was therefore extended--from peripheral vascular disease and connective tissue diseases to the whole spectrum of trophic changes in the skin of the extremities. Systems for digital image analysis allow quantification of the structure of the microvascular bed (quantitative appraisal of microangiopathies) and function (capillary haemodynamics and exchange). Laser Doppler and capillaroscopy can also be combined for the measurement of red blood cell velocity in single capillaries.

Humans↗

PubMed1999

New techniques for clinical assessment of the peripheral microcirculation.

Blood Flow Velocity↗

PubMed1999

Dynamic digital fluorescence ratio imaging of cell calcium in vascular endothelial cells.

AIM: To study the spatial and temporal distribution of intracellular Ca2+ concentration in cultured bovine pulmonary artery endothelial (BPAE) cells. METHODS: Cultured BPAE cells were loaded with Fura-2 and observed under an inverted microscope coupled to a microfluorimeter, which enables pixel-to-pixel ratio imaging of the BPAE cells in real time. RESULTS: Addition of Ca2+ 1-2 mmol.L-1 to BPAE cells, which were exposed to Ca(2+)-free medium containing egtazic acid, resulted in a transient elevation of cytosolic Ca2+ concentration, which rapidly returned to the resting level. Biphasic elevation (a larger transient phase followed by a smaller sustained phase) of intracellular Ca2+ concentration was observed upon the addition of ATP (via activation of surface membrane receptor). 4-Chloro-3-ethyl phenol (CEP; an activator of Ca(2+)-induced Ca2+ channels) potently induced elevation of Ca2+ level. Cyclopiazonic acid (CPA; an inhibitor of endoplasmic reticulum Ca(2+)-ATPase pump) offered a more sustained elevation of Ca2+. In most cases, the highest level of Ca2+ elevation was observed around the cell peripheries, sometimes at rest and particularly upon stimulation. Ca2+ elevation associated with nuclear complex seemed to be higher compared to that in the cytosolic compartment. CONCLUSION: Changes of cell Ca2+ upon stimulation by various agents that acted at different intracellular sites were found to be temporarily and spatially heterogenous among BPAE cells. At the single cell level, Ca2+ elevation seemed to occur initially near the peripheral region followed by the nuclear region. This study raised the possibility that nuclear Ca2+ and cytosolic Ca2+ might be regulated independently in BPAE cells.

Animals↗