Simultaneous measurements of Ca2+ currents and intracellular Ca2+ concentrations in single skeletal muscle fibers of the frog

1987 ◽  
Vol 65 (4) ◽  
pp. 681-685 ◽  
Author(s):  
G. Brum ◽  
E. Stefani ◽  
E. Rios
Keyword(s):  
Time Course ◽  
Muscle Fibers ◽  
Optical Signal ◽  
Single Muscle ◽  
Drastic Reduction ◽  
Maximum Value ◽  
Input Flux ◽  
Voltage Dependent ◽  
The Relationship ◽  
Removal Model

The relationship between Ca2+ current amplitudes and myoplasmic Ca2+ transients was studied in single muscle fibers. Segments of muscle fibers were voltage-clamped in a double Vaseline gap chamber. Ca2+ transients were measured as an optical signal derived from the interaction between Ca2+ and the dye antipyrylazo III. The cells were maintained at −90 mV. Ca2+ currents were detected at pulse potentials to −50 mV, reached a maximum value at 0 mV, were reduced in size for larger depolarizations, and reversed at about 40 mV. Ca2+ transients were also detected at −50 Mv and progressively increased in size with larger pulse potentials up to 10 mV. Depolarizations to voltages greater than 10 mV did not further increase the size of the transient. The magnitude and time course of transients from 10 to 70 mV were almost identical. Ca2+ fluxes into the myoplasm (Ca2+ input fluxes) were calculated from the Ca2+ transients applying a removal model. The size of the input fluxes increased with depolarization up to 0 mV. Between 0 and 70 mV the peak input flux slightly increased, while the flux measured at 200 ms remained unchanged. In conclusion, Ca2+ transients and input fluxes were not reduced during pulses to large positive potentials, even though a drastic reduction of Ca2+ current occurred at these potentials. These observations make it very unlikely that a voltage-dependent Ca2+ entry is the triggering signal for contraction.

scholarly journals Effects of Caffeine on Crayfish Muscle Fibers

1970 ◽  
Vol 55 (5) ◽  
pp. 640-664 ◽  
Author(s):  
Dante J. Chiarandini ◽  
John P. Reuben ◽  
Philip W. Brandt ◽  
Harry Grundfest
Keyword(s):  
Time Course ◽  
Muscle Fibers ◽  
Resting Potential ◽  
Induced Responses ◽  
Single Muscle ◽  
Transverse Tubular System ◽  
Crayfish Muscle ◽  
Number Of Factors ◽  
Tubular System

Contractions are evoked in single muscle fibers of crayfish by intracellular as well as extracellular applications of caffeine. Responses to external applications in concentrations above 2 mM could be induced indefinitely. With concentrations above 5 mM the caffeine-induced responses were highly repeatable. Tensions were transient even when the caffeine remained in the bath. There was no change in resting potential, but during the contraction the effective resistance decreased about 10%. A number of factors (change in pH, Ca, K, and Cl) modified the responses. The time course of the tension was greatly prolonged when the transverse tubular system (TTS) was s swollen and was again shortened when the TTS was caused to shrink. An increased permeability to Ca induced by caffeine was evidenced by the transformation of the normally graded electrical responses to Ca spikes, which are insensitive to tetrodotoxin. The overshoot is a function of both external Ca and caffeine. A 10-fold change in Ca changed the overshoot by 19 mv in the presence of 10 mM caffeine and by 29 mv in 80 mM caffeine. The role of the increased permeability to Ca for caffeine-induced contractions will be analyzed in the accompanying paper.

scholarly journals Voltage-dependent Ca2+ Fluxes in Skeletal Myotubes Determined Using a Removal Model Analysis

2003 ◽  
Vol 123 (1) ◽  
pp. 33-52 ◽  
Author(s):  
R.P. Schuhmeier ◽  
W. Melzer
Keyword(s):  
Time Course ◽  
Voltage Dependence ◽  
Flux Ratio ◽  
Cyclopiazonic Acid ◽  
High Concentration ◽  
Fura 2 ◽  
Input Flux ◽  
Indicator Dye ◽  
Removal Model ◽  
Tonic Phase

The purpose of this study was to quantify the Ca2+ fluxes underlying Ca2+ transients and their voltage dependence in myotubes by using the “removal model fit” approach. Myotubes obtained from the mouse C2C12 muscle cell line were voltage-clamped and loaded with a solution containing the fluorescent indicator dye fura-2 (200 μM) and a high concentration of EGTA (15 mM). Ca2+ inward currents and intracellular ratiometric fluorescence transients were recorded in parallel. The decaying phases of Ca2+-dependent fluorescence signals after repolarization were fitted by theoretical curves obtained from a model that included the indicator dye, a slow Ca2+ buffer (to represent EGTA), and a sequestration mechanism as Ca2+ removal components. For each cell, the rate constants of slow buffer and transport and the off rate constant of fura-2 were determined in the fit. The resulting characterization of the removal properties was used to extract the Ca2+ input fluxes from the measured Ca2+ transients during depolarizing pulses. In most experiments, intracellular Ca2+ release dominated the Ca2+ input flux. In these experiments, the Ca2+ flux was characterized by an initial peak followed by a lower tonic phase. The voltage dependence of peak and tonic phase could be described by sigmoidal curves that reached half-maximal activation at −16 and −20 mV, respectively, compared with −2 mV for the activation of Ca2+ conductance. The ratio of the peak to tonic phase (flux ratio) showed a gradual increase with voltage as in rat muscle fibers indicating the similarity to EC coupling in mature mammalian muscle. In a subgroup of myotubes exhibiting small fluorescence signals and in cells treated with 30 μM of the SERCA pump inhibitor cyclopiazonic acid (CPA) and 10 mM caffeine, the calculated Ca2+ input flux closely resembled the L-type Ca2+ current, consistent with the absence of SR Ca2+ release under these conditions and in support of a valid determination of the time course of myoplasmic Ca2+ input flux based on the optical indicator measurements.

scholarly journals Detection of Ca2+ transients near ryanodine receptors by targeting fluorescent Ca2+ sensors to the triad

2021 ◽  
Vol 153 (4) ◽  
Author(s):  
Colline Sanchez ◽  
Christine Berthier ◽  
Yves Tourneur ◽  
Laloé Monteiro ◽  
Bruno Allard ◽  
...  
Keyword(s):  
Time Course ◽  
Model Simulation ◽  
Ryanodine Receptors ◽  
Muscle Fibers ◽  
Contrast Model ◽  
Spatial Gradients ◽  
Lower Amplitude ◽  
Voltage Dependent ◽  
Intact Muscle ◽  
Spatial Level

In intact muscle fibers, functional properties of ryanodine receptor (RYR)–mediated sarcoplasmic reticulum (SR) Ca2+ release triggered by activation of the voltage sensor CaV1.1 have so far essentially been addressed with diffusible Ca2+-sensitive dyes. Here, we used a domain (T306) of the protein triadin to target the Ca2+-sensitive probe GCaMP6f to the junctional SR membrane, in the immediate vicinity of RYR channels, within the triad region. Fluorescence of untargeted GCaMP6f was distributed throughout the muscle fibers and experienced large Ca2+-dependent changes, with obvious kinetic delays, upon application of voltage-clamp depolarizing pulses. Conversely, T306-GCaMP6f localized to the triad and generated Ca2+-dependent fluorescence transients of lower amplitude and faster kinetics for low and intermediate levels of Ca2+ release than those of untargeted GCaMP6f. By contrast, model simulation of the spatial gradients of Ca2+ following Ca2+ release predicted limited kinetic differences under the assumptions that the two probes were present at the same concentration and suffered from identical kinetic limitations. At the spatial level, T306-GCaMP6f transients within distinct regions of a same fiber yielded a uniform time course, even at low levels of Ca2+ release activation. Similar observations were made using GCaMP6f fused to the γ1 auxiliary subunit of CaV1.1. Despite the probe's limitations, our results point out the remarkable synchronicity of voltage-dependent Ca2+ release activation and termination among individual triads and highlight the potential of the approach to visualize activation or closure of single groups of RYR channels. We anticipate targeting of improved Ca2+ sensors to the triad will provide illuminating insights into physiological normal RYR function and its dysfunction under stress or pathological conditions.

scholarly journals Frozen/thawed meat quality associated with muscle fiber characteristics of porcine longissimus thoracis et lumborum, psoas major, semimembranosus, and semitendinosus muscles

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Huilin Cheng ◽  
Sumin Song ◽  
Gap-Don Kim
Keyword(s):  
Meat Quality ◽  
Muscle Fiber ◽  
Shear Force ◽  
Muscle Fibers ◽  
Frozen Storage ◽  
Meat Color ◽  
Pork Meat ◽  
Psoas Major ◽  
The Relationship

AbstractTo evaluate the relationship between muscle fiber characteristics and the quality of frozen/thawed pork meat, four different muscles, M. longissimus thoracis et lumborum (LTL), M. psoas major (PM), M. semimembranosus (SM), and M. semitendinosus (ST), were analyzed from twenty carcasses. Meat color values (lightness, redness, yellowness, chroma, and hue) changed due to freezing/thawing in LTL, which showed larger IIAX, IIX, and IIXB fibers than found in SM (P < 0.05). SM and ST showed a significant decrease in purge loss and an increase in shear force caused by freezing/thawing (P < 0.05). Compared with LTL, SM contains more type IIXB muscle fibers and ST had larger muscle fibers I and IIA (P < 0.05). PM was the most stable of all muscles, since only its yellowness and chroma were affected by freezing/thawing (P < 0.05). These results suggest that pork muscle fiber characteristics of individual cuts must be considered to avoid quality deterioration during frozen storage.

Computer Simulation of the Responses of Human Motoneurons to Composite 1A EPSPS: Effects of Background Firing Rate

1997 ◽  
Vol 77 (1) ◽  
pp. 405-420 ◽  
Author(s):  
Kelvin E. Jones ◽  
Parveen Bawa
Keyword(s):  
Computer Simulation ◽  
Firing Rate ◽  
Time Course ◽  
Current Model ◽  
Response Probability ◽  
Muscle Spindles ◽  
Excitatory Postsynaptic Potential ◽  
Biophysical Properties ◽  
Rhythmic Firing ◽  
The Relationship

Jones, Kelvin E. and Parveen Bawa. Computer simulation of the responses of human motoneurons to composite 1A EPSPS: effects of background firing rate. J. Neurophysiol. 77: 405–420, 1997. Two compartmental models of spinal alpha motoneurons were constructed to explore the relationship between background firing rate and response to an excitatory input. The results of these simulations were compared with previous results obtained from human motoneurons and discussed in relation to the current model for repetitively firing human motoneurons. The morphologies and cable parameters of the models were based on two type-identified cat motoneurons previously reported in the literature. Each model included five voltage-dependent channels that were modeled using Hodgkin-Huxley formalism. These included fast Na+ and K+ channels in the initial segment and fast Na+ and K+ channels as well as a slow K+ channel in the soma compartment. The density and rate factors for the slow K+ channel were varied until the models could reproduce single spike AHP parameters for type-identified motoneurons in the cat. Excitatory synaptic conductances were distributed along the equivalent dendrites with the same density described for la synapses from muscle spindles to type-identified cat motoneurons. Simultaneous activation of all synapses on the dendrite resulted in a large compound excitatory postsynaptic potential (EPSP). Brief depolarizing pulses injected into a compartment of the equivalent dendrite resulted in pulse potentials (PPs), which resembled the compound EPSPs. The effects of compound EPSPs and PPs on firing probability of the two motoneuron models were examined during rhythmic firing. Peristimulus time histograms, constructed between the stimulus and the spikes of the model motoneuron, showed excitatory peaks whose integrated time course approximated the time course of the underlying EPSP or PP as has been shown in cat motoneurons. The excitatory peaks were quantified in terms of response probability, and the relationship between background firing rate and response probability was explored. As in real human motoneurons, the models exhibited an inverse relationship between response probability and background firing rate. The biophysical properties responsible for the relationship between response probability and firing rate included the shapes of the membrane voltage trajectories between spikes and nonlinear changes in PP amplitude during the interspike interval at different firing rates. The results from these simulations suggest that the relationship between response probability and background firing rate is an intrinsic feature of motoneurons. The similarity of the results from the models, which were based on the properties of cat motoneurons, and those from human motoneurons suggests that the biophysical properties governing rhythmic firing in human motoneurons are similar to those of the cat.

Metric for Disassembly and Reuse Decisions: Formulation and Validation

2008 ◽  
Author(s):  
Vijitashwa Pandey ◽  
Deborah Thurston
Keyword(s):  
Maximum Likelihood ◽  
Value Function ◽  
Choice Theory ◽  
Decision Makers ◽  
Maximum Likelihood Estimates ◽  
Likelihood Method ◽  
Maximum Value ◽  
Long Range Planning ◽  
The Relationship ◽  
The Value Function

Design for disassembly and reuse focuses on developing methods to minimize difficulty in disassembly for maintenance or reuse. These methods can gain substantially if the relationship between component attributes (material mix, ease of disassembly etc.) and their likelihood of reuse or disposal is understood. For products already in the marketplace, a feedback approach that evaluates willingness of manufacturers or customers (decision makers) to reuse a component can reveal how attributes of a component affect reuse decisions. This paper introduces some metrics and combines them with ones proposed in literature into a measure that captures the overall value of a decision made by the decision makers. The premise is that the decision makers would choose a decision that has the maximum value. Four decisions are considered regarding a component’s fate after recovery ranging from direct reuse to disposal. A method on the lines of discrete choice theory is utilized that uses maximum likelihood estimates to determine the parameters that define the value function. The maximum likelihood method can take inputs from actual decisions made by the decision makers to assess the value function. This function can be used to determine the likelihood that the component takes a certain path (one of the four decisions), taking as input its attributes, which can facilitate long range planning and also help determine ways reuse decisions can be influenced.

scholarly journals Excessive Pressure in Multichambered Cuffs Used for Sequential Compression Therapy

2002 ◽  
Vol 82 (10) ◽  
pp. 1000-1008 ◽  
Author(s):  
Patrick Segers ◽  
Jean-Paul Belgrado ◽  
Andre Leduc ◽  
Olivier Leduc ◽  
Pascal Verdonck
Keyword(s):  
Time Course ◽  
Compression Therapy ◽  
Lymphatic Drainage ◽  
Chamber Pressure ◽  
Excessive Pressure ◽  
Ellipsoidal Model ◽  
Order Of Magnitude ◽  
Pneumatic Compression Devices ◽  
Target Pressure ◽  
The Relationship

Abstract Background and Purpose. Pneumatic compression devices, used as part of the therapeutic strategy for lymphatic drainage, often have cuffs with multiple chambers that are inflated sequentially. The purpose of this study was to investigate (1) the relationship between cuff chamber pressure (Pchamber) and the pressure on the cuff-skin interface (Pinterface) and (2) the mechanical interaction of cuff chambers and consequences for device control. Subjects and Methods. In this study, we used 3 cylindrical (60-, 80-, and 100-mm-diameter) model limbs and 1 ellipsoidal model of the arm to test a commercially available pressure controller using “target pressures,” indicated by the controller, of 30, 60, 80, and 100 mm Hg. We studied the time course of Pchamber and Pinterface during the inflation sequence and the effect of local curvature on Pinterface. Results. Our data indicated that, overall, Pinterface is of the same order of magnitude as Pchamber. There was some effect of model diameter and shape, with the smaller curvatures yielding the highest Pinterface. Cuff chamber interaction led to Pchamber and Pinterface values in the most distal (first inflated) chamber that were up to 80% higher than the target pressure. For the 80-mm cylindrical model, for instance, pressure in this chamber reached 54, 98, 121, and 141 mm Hg, respectively, instead of the 30, 60, 80, and 100 mm Hg indicated by the controller. Discussion and Conclusion. The discrepancy between the target pressure, indicated by the controller, and the pressure measured inside the cuff chambers undermines the therapeutic control and efficacy of the pneumatic compression devices. Because the measured pressures were far beyond the pressure level indicated by the controller, it is recommended that pneumatic compression devices be used at much lower target pressures (&lt;30 mm Hg) than those applied in clinical practice.

ANALYSIS OF THE ERROR IN DETERMINING THE EQUIVALENT SIZE OF A DEFECT BY THE STANDARDLESS METHOD DURING ULTRASONIC TESTING

2021 ◽  
pp. 50-57
Author(s):  
A. N. Kireev ◽  
M. A. Kireeva
Keyword(s):  
Relative Error ◽  
High Reliability ◽  
Ultrasonic Method ◽  
Experimental Studies ◽  
Standard Samples ◽  
Defect Identification ◽  
Identification Method ◽  
Maximum Value ◽  
Software Product ◽  
The Relationship

The article provides a review and analysis of the defect identification method for determining the size of discontinuities when diagnosing various machine parts and units by the manual ultrasonic method. This method makes it possible to determine the equivalent size of discontinuities of various types without using standard samples of an enterprise: point planar and volumetric; extended planar and volumetric. The method is based on the use of the relationship between the amplitude and time characteristics of the echo signal from the discontinuity and the backside signal in the object being diagnosed and the equivalent size of the discontinuity. The article presents the mathematical apparatus for the implementation of this method. Also presented is a software product that allows you to automate calculations when using this defect identification method. The article contains experimental studies of the method for determining the equivalent dimensions of discontinuities of various types, which have shown its high reliability. The maximum value of the relative error in determining the equivalent size of a point planar discontinuity was 2.867 %. The maximum value of the relative error in determining the equivalent size of a point volumetric discontinuity was 1.986 %. The maximum value of the relative error in determining the transverse equivalent size of an extended planar discontinuity was 0.667 %. The maximum value of the relative error in determining the transverse equivalent size of an extended volumetric discontinuity was 1.95 %.

scholarly journals New Insights on the Mechanism of Fatty Acids as Buccal Permeation Enhancers

Pharmaceutics ◽  
2018 ◽  
Vol 10 (4) ◽  
pp. 201 ◽  
Author(s):  
Cristina Padula ◽  
Silvia Pescina ◽  
Sara Nicoli ◽  
Patrizia Santi
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Fluorescein Isothiocyanate ◽  
Penetration Enhancers ◽  
Systemic Delivery ◽  
Permeation Enhancers ◽  
Maximum Value ◽  
The Relationship ◽  
Insight Into ◽  
The Ideal

Buccal mucosa has recently received much attention as a potential route for systemic delivery of drugs, including biologics and vaccines. The aim of this work was to gain insight into the mechanism of fatty acids as buccal permeation enhancers, by studying the effect of a series of medium and long chain fatty acids on the permeation of a model high molecular weight and hydrophilic molecule, fluorescein isothiocyanate labelled dextran (FD-4, m.w. 4 kDa) across porcine esophageal epithelium. A parabolic relationship between fatty acid lipophilicity and enhancement was obtained, regardless of the presence and number of double bonds. The relationship, which resembles the well-known relationship between permeability and lipophilicity of transdermal delivery, presents a maximum value in correspondence of C10 (logP approx. 4). This is probably the ideal lipophilicity for the fatty acid to interact with the lipid domains of the mucosa. When the same analysis was performed on skin data, the same trend was observed, although the maximum value was reached for C12 (logP approx. 5), in agreement with the higher lipophilicity of the skin. The results obtained in the present work represent a significant advancement in the understanding of the mechanisms of action of fatty acids as buccal penetration enhancers.

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