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1.
Critical and supercritical spatiotemporal calcium dynamics in beta cells
Marko Gosak, Andraž Stožer, Rene Markovič, Jurij Dolenšek, Matjaž Perc, Marjan Rupnik, Marko Marhl, 2017, izvirni znanstveni članek

Opis: A coordinated functioning of beta cells within pancreatic islets is mediated by oscillatory membrane depolarization and subsequent changes in cytoplasmic calcium concentration. While gap junctions allow for intraislet information exchange, beta cells within islets form complex syncytia that are intrinsically nonlinear and highly heterogeneous. To study spatiotemporal calcium dynamics within these syncytia, we make use of computational modeling and confocal high-speed functional multicellular imaging. We show that model predictions are in good agreement with experimental data, especially if a high degree of heterogeneity in the intercellular coupling term is assumed. In particular, during the first few minutes after stimulation, the probability distribution of calcium wave sizes is characterized by a power law, thus indicating critical behavior. After this period, the dynamics changes qualitatively such that the number of global intercellular calcium events increases to the point where the behavior becomes supercritical. To better mimic normal in vivo conditions, we compare the described behavior during supraphysiological non-oscillatory stimulation with the behavior during exposure to a slightly lower and oscillatory glucose challenge. In the case of this protocol, we observe only critical behavior in both experiment and model. Our results indicate that the loss of oscillatory changes, along with the rise in plasma glucose observed in diabetes, could be associated with a switch to supercritical calcium dynamics and loss of beta cell functionality.
Ključne besede: beta cells, islets of Langerhans, self-organized criticality, intercellular dynamics, calcium waves, glucose oscillations, computational model, confocal calcium imaging
Objavljeno v DKUM: 23.01.2018; Ogledov: 999; Prenosov: 328
.pdf Celotno besedilo (3,43 MB)
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2.
Progressive glucose stimulation of islet beta cells reveals a transition from segregated to integrated modular functional connectivity patterns
Rene Markovič, Andraž Stožer, Marko Gosak, Jurij Dolenšek, Marko Marhl, Marjan Rupnik, 2015, izvirni znanstveni članek

Opis: Collective beta cell activity in islets of Langerhans is critical for the supply of insulin within an organism. Even though individual beta cells are intrinsically heterogeneous, the presence of intercellular coupling mechanisms ensures coordinated activity and a well-regulated exocytosis of insulin. In order to get a detailed insight into the functional organization of the syncytium, we applied advanced analytical tools from the realm of complex network theory to uncover the functional connectivity pattern among cells composing the intact islet. The procedure is based on the determination of correlations between long temporal traces obtained from confocal functional multicellular calcium imaging of beta cells stimulated in a stepwise manner with a range of physiological glucose concentrations. Our results revealed that the extracted connectivity networks are sparse for low glucose concentrations, whereas for higher stimulatory levels they become more densely connected. Most importantly, for all ranges of glucose concentration beta cells within the islets form locally clustered functional sub-compartments, thereby indicating that their collective activity profiles exhibit a modular nature. Moreover, we show that the observed non-linear functional relationship between different network metrics and glucose concentration represents a well-balanced setup that parallels physiological insulin release.
Ključne besede: endocrinology, computational biophysics, calcium signalling, biological physics
Objavljeno v DKUM: 23.06.2017; Ogledov: 935; Prenosov: 282
.pdf Celotno besedilo (957,14 KB)
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3.
Membrane potential and calcium dynamics in beta cells from mouse pancreas tissue slices : theory, experimentation, and analysis
Jurij Dolenšek, Denis Špelič, Maša Skelin, Borut Žalik, Marko Gosak, Marjan Rupnik, Andraž Stožer, 2015, pregledni znanstveni članek

Opis: Beta cells in the pancreatic islets of Langerhans are precise biological sensors for glucose and play a central role in balancing the organism between catabolic and anabolic needs. A hallmark of the beta cell response to glucose are oscillatory changes of membrane potential that are tightly coupled with oscillatory changes in intracellular calcium concentration which, in turn, elicit oscillations of insulin secretion. Both membrane potential and calcium changes spread from one beta cell to the other in a wave-like manner. In order to assess the properties of the abovementioned responses to physiological and pathological stimuli, the main challenge remains how to effectively measure membrane potential and calcium changes at the same time with high spatial and temporal resolution, and also in as many cells as possible. To date, the most wide-spread approach has employed the electrophysiological patch-clamp method to monitor membrane potential changes. Inherently, this technique has many advantages, such as a direct contact with the cell and a high temporal resolution. However, it allows one to assess information from a single cell only. In some instances, this technique has been used in conjunction with CCD camera-based imaging, offering the opportunity to simultaneously monitor membrane potential and calcium changes, but not in the same cells and not with a reliable cellular or subcellular spatial resolution. Recently, a novel family of highly-sensitive membrane potential reporter dyes in combination with high temporal and spatial confocal calcium imaging allows for simultaneously detecting membrane potential and calcium changes in many cells at a time. Since the signals yielded from both types of reporter dyes are inherently noisy, we have developed complex methods of data denoising that permit for visualization and pixel-wise analysis of signals. Combining the experimental approach of high-resolution imaging with the advanced analysis of noisy data enables novel physiological insights and reassessment of current concepts in unprecedented detail.
Ključne besede: calcium sensors, membrane potential sensors, calcium imaging, membrane potential imaging, beta cell, pancreas, denoising, patch-clamp
Objavljeno v DKUM: 22.06.2017; Ogledov: 983; Prenosov: 151
.pdf Celotno besedilo (4,17 MB)
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4.
The analysis of intracellular and intercellular calcium signaling in human anterior lens capsule epithelial cells with regard to different types and stages of the cataract
Marko Gosak, Rene Markovič, Aleš Fajmut, Marko Marhl, Marko Hawlina, Sofija Andjelić, 2015, izvirni znanstveni članek

Opis: In this work we investigated how modifications of the Ca2+ homeostasis in anterior lens epithelial cells (LECs) are associated with different types of cataract (cortical or nuclear) and how the progression of the cataract (mild or moderate) affects the Ca2+ signaling. We systematically analyzed different aspects of intra- and inter-cellular Ca2+ signaling in the human LECs, which are attached to surgically isolated lens capsule (LC), obtained during cataract surgery. We monitored the temporal and spatial changes in intracellular Ca2+ concentration after stimulation with acetylcholine by means of Fura-2 fluorescence captured with an inverted microscope. In our analysis we compared the features of Ca2+ signals in individual cells, synchronized activations, spatio-temporal grouping and the nature of intercellular communication between LECs. The latter was assessed by using the methodologies of the complex network theory. Our results point out that at the level of individual cells there are no significant differences when comparing the features of the signals with regard either to the type or the stage of the cataract. On the other hand, noticeable differences are observed at the multicellular level, despite inter-capsule variability. LCs associated with more developed cataracts were found to exhibit a slower collective response to stimulation, a less pronounced spatio-temporal clustering of LECs with similar signaling characteristics. The reconstructed intercellular networks were found to be sparser and more segregated than in LCs associated with mild cataracts. Moreover, we show that spontaneously active LECs often operate in localized groups with quite well aligned Ca2+ activity. The presence of spontaneous activity was also found to affect the stimulated Ca2+ responses of individual cells. Our findings indicate that the cataract progression entails the impairment of intercellular signaling thereby suggesting the functional importance of altered Ca2+ signaling of LECs in cataractogenesis.
Ključne besede: intracellular calcium signaling, anterior lens, epithelial cells, cataract
Objavljeno v DKUM: 19.06.2017; Ogledov: 889; Prenosov: 373
.pdf Celotno besedilo (1,45 MB)
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5.
The influence of physico-chemical parameters on water scale precipitation on washing machines' heaters
Danijela Urbancl, Darko Goričanec, Jurij Krope, 2007, strokovni članek

Opis: The paper presents the results of analysis of physico - chemical parameters influence on the intensity of water scale precipitation on washing machines heaters. Washing machines of the same producer were, to this end, modified to an endless cycle of washing with the same amount of cotton fabric at the same conditions. On the basis of various experiments it was determined that water hardness, the amount of exceeded carbon dioxide, water turbulence around the heater, the amount of cotton fibres in water exceeded from fabrics, and heater specific heat strength have the key role in water scale building up in washing machines heaters.
Ključne besede: magnetic water treatment, magnetic hydrodynamics, water scale, calcium carbonate, scale prevention, washing machine, electrical heater
Objavljeno v DKUM: 21.12.2015; Ogledov: 1173; Prenosov: 99
.pdf Celotno besedilo (544,97 KB)
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6.
Modeling of molecular and cellular mechanisms involved in [Ca sup 2+] signal encoding in airway myocytes
Marko Marhl, Denis Noble, Etienne Roux, 2006, pregledni znanstveni članek

Opis: In airway myocytes signal transduction via cytosolic calcium plays an important role. In relation with experimental results we review models of basic molecular and cellular mechanisms involved in the signal transduction from the myocyte stimulation to the activation of the contractile apparatus. We concentrate on mechanisms for encoding of input signals into Ca2+ signals and the mechanisms for their decoding. The mechanisms are arranged into a general scheme of cellular signaling, the so-called bow-tie architecture of signaling, in which calcium plays the role of a common media for cellular signals and links the encoding and decoding part. The encoding of calcium signals in airway myocytes is better known and is presented in more detail. Inparticular, we focus on three recent models taking into account the intracellular calcium handling and ion fluxes through the plasma membrane. Themodel of membrane conductances was originally proposed for predicting membrane depolarization and voltage-dependent Ca2+ influx triggered by initialcytosolic Ca2+ increase as observed on cholinergic stimulation. Cellular models of intracellular Ca2+ handling were developed to investigate the role of a mixed population of InsP3 receptor isoforms and the cellular environment in the occurrence of Ca2+ oscillations, and the respective role ofthe sarcoplasmic reticulum, mitochondria, and cytosolic Ca2+-binding proteins in cytosolic Ca2+ clearance. Modeling the mechanisms responsible for the decoding of calcium signals is developed in a lesser extent; however, the most recent theoretical studies are briefly presented in relation with the known experimental results.
Ključne besede: biophysics, mathematical modelling, modelling, calcium oscillations, contractions, airway smooth muscle cells, muscle cells, smooth muscles, encoding, decoding, bow-tie structures
Objavljeno v DKUM: 07.06.2012; Ogledov: 1555; Prenosov: 31
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7.
Theoretical and experimental investigation of calcium-contraction coupling in airway smooth muscle
Prisca Mbikou, Aleš Fajmut, Milan Brumen, Etienne Roux, 2006, izvirni znanstveni članek

Opis: We investigated theoretically and experimentally the ▫$Ca^{2+}$▫-contraction couplingin rat tracheal smooth muscle. ▫$[Ca^{2+}]_i$▫, isometric contraction and myosin light chain (MLC) phosphorylation were measured in response to 1 mM carbachol. Theoretical modeling consisted in coupling a model of ▫$Ca^{2+}-dependent$▫ MLC kinase (MLCK) activation with a four-state model of smooth muscle contractile apparatus. Stimulation resulted in a short-time contraction obtained within 1 min, followed by a long-time contraction up to the maximal force obtained in 30 min. ML-7 and Wortmannin (MLCK inhibitors) abolished the contraction. Chelerythrine (PKC inhibitor) did not change the short-time, but reduced the long-time contraction. ▫$[Ca^{2+}]_i$▫ responses of isolated myocytes recorded during the first 90 s consisted in a fast peak, followed by a plateau phase and, in 28 % of the cells, superimposed ▫$Ca^{2+}$▫ oscillations. MLC phosphorylation was maximal at 5 s and then decreased, whereas isometric contraction followed a Hill-shaped curve. The model properlypredicts the time course of MLC phosphorylation and force of the short-time response. With oscillating ▫$Ca^{2+}$▫ signal, the predicted force does not oscillate. According to the model, the amplitude of the plateau and the frequency of oscillations encode for the amplitude of force, whereas the peak encodes for force velocity. The long-time phase of the contraction, associated with a second increase in MLC phosphorylation, may be explained, at least partially, by MLC phosphatase (MLCP) inhibition, possibly via PKC inhibition.
Ključne besede: biophysics, mathematical modelling, modelling, calcium oscillations, contractions, force development, muscle cells, smooth muscles, myosin kinase
Objavljeno v DKUM: 07.06.2012; Ogledov: 1463; Prenosov: 82
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8.
Selective regulation of protein activity by complex Ca[sup]2+ oscillations : a theoretical study
Beate Knoke, Marko Marhl, Stefan Schuster, 2007, samostojni znanstveni sestavek ali poglavje v monografski publikaciji

Opis: Calcium oscillations play an important role in intracellular signal transduction. As a second messenger, ▫$Ca^{2+}$▫ represents a link between several input signals and several target processes in the cell. Whereas the frequency of simple ▫$Ca^{2+}$▫ oscillations enables a selective activation of a specific protein and herewith a particular process, the question arises of how at the same time two or more classes of proteins can be specifically regulated. The question is general and concerns the problem of how one second messenger can transmit more than one signal simultaneously (bow-tie structure of signalling). To investigate whether a complex ▫$Ca^{2+}$▫ signal like bursting, a succession of low-peak and high-peak oscillatory phases, could selectively activate different proteins, several bursting patterns with simplified square pulses were applied in a theoretical model. The results indicate that bursting ▫$Ca^{2+}$▫ oscillations allow a differential regulation of two different calcium-binding proteins, and hence, perform the desired function.
Ključne besede: biophysics, calcium oscillations, cellular dynamics, mathematical models, signalling, bow-tie structures, bursting, decoding
Objavljeno v DKUM: 07.06.2012; Ogledov: 1461; Prenosov: 23
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9.
Noise-induced spatial dynamics in the presence of memory loss
Matjaž Perc, Marko Marhl, 2007, izvirni znanstveni članek

Opis: We study the spatial dynamics of noise-induced waves in two-dimensional excitable media in dependence on the duration of the artificially imposed refractory time that is introduced to each constitutive system unit after an excitation. Due to the introduction of refractory times, a randomly induced spatial wave is temporarily unable to transmit information to the opposite site of its propagation direction. Thus, once the wave leaves the absorbing boundaries of the spatial grid the system has little or no recollection, depending on the duration of the refractory time, of its existence. We show that even in the presence of such memory loss, self-organization of excitatory events leads to noise-induced spatial periodicity in the media. We present a simple analytical treatment of a two-unit system to capture and explain the essence of the observed phenomenon. Since refractory times are widespread in biological systems, our results provide interesting insights into functioning of real-life organisms at the cellular as well as tissue level.
Ključne besede: noise, spatiotemporal noise, intensity, pattern formation, refractory time, calcium oscillations
Objavljeno v DKUM: 07.06.2012; Ogledov: 1588; Prenosov: 84
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10.
Role of cascades in converting oscillatory signals into stationary step-like responses
Marko Marhl, Vladimir Grubelnik, 2007, izvirni znanstveni članek

Opis: In biological signal transduction pathways intermediates are often oscillatory and need to be converted into smooth output signals at the end. We show by mathematical modelling that protein kinase cascades enable converting oscillatory signals into sharp stationary step-like outputs. The importance of this result is demonstrated for the switch-like protein activation by calcium oscillations, which is of biological importance for regulating different cellular processes. In addition, we found that protein kinase cascades cause memory effects in the protein activation, which might be of a physiological advantage since a smaller amount of calcium transported in the cell is required for an effective activation of cellular processes.
Ključne besede: physics, calcium oscillations, mathematical modelling, calcium, calcium oscillations, sygnalling cascade, protein kinase cascades, signal transduction, ultrasensitivity, biochemical switch, cellular dynamics
Objavljeno v DKUM: 07.06.2012; Ogledov: 1626; Prenosov: 77
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