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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 9 (1972), S. 341-360 
    ISSN: 1432-1424
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary The passive electrical properties of neonatal rat heart cells grown in monolayer cultures were determined. Hyperpolarizing current pulses were injected through one microelectrode via an active bridge circuit. Membrane voltage displacements caused by the injected current pulses were measured at various distances from the first with a second microelectrode. Using a modified least-squares method the experimental results were fitted to a Bessel function, which is the steady-state solution of the differential equation describing the relation between membrane voltage caused by current injection and interelectrode distance in a very large and very thin plane cell. Best fit was obtained with a space constant of 360 μm and an internal resistivity of 500 Ω cm. From these figures, specific membrane resistance was calculated to be 1,300 Ω cm2, assuming all current to leave through the upper surface of the monolayer. The time constant of the membrane was measured from the time course of the current-induced membrane voltage displacements. From its value of 1.7 msec a membrane capacity of 1.3 μF/cm2 was calculated. From these results and some literature data on nexus distribution (A. W. Spira,J. Ultrastruct. Res. 34:409, 1971) specific nexus resistance was calculated to range between 0.25 and 1.25 Ω cm2, depending on the amount of folding of the intercalated discs. The results suggest that spread of activation in monolayer cultures of heart cells by means of local circuit currents is very likely.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-1424
    Keywords: beat-rate synchronization ; morphometry ; junctional conductance ; voltage clamp ; single-channel conductance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary The time course of gap junction formation and growth, following contraction synchronization of cardiac myocytes in culture, has been studied in a combined (electro)physiological and ultrastructural study. In cultures of collagenase-dissociated neonatal rat cardiocytes, pairs of spontaneously beating myocytes synchronized their contractions within one beat interval within 2–20 min after they apparently had grown into contact, 45 sec after the first synchronized beat an appreciable junctional region containing several small gap junctions was already present. In the following 30 min, neither the area of individual gap junctions nor their total area increased, 75 min after synchronization both the area of individual gap junctions and their total area had increased by a factor of 10–15 with respect to what was found in the first half hour. In the period between 75 and 300 min again no further increase in gap junctional area was found. In double voltage-clamp experiments, gap junctions between well-coupled cells behaved like ohmic conductors. In poorly coupled cells, in which the number of functional gap-junctional channels was greatly reduced, the remaining channels showed voltage-dependent gating. Their single-channel conductance was 40–50 pS. The electrophysiologically measured junctional conductance agreed well with the conductance calculated from the morphometrically determined gap-junctional area. It is concluded that a rapid initial gap junction formation occurs during the 2–20 min period prior to synchronization by assembly of functional channels from existing channel precursors already present in the cell membranes. It then takes at least another 30 min before the gap-junctional area increases possibly byde novo synthesis or by recruitment from intracellular stores or from nonjunctional membranes, a process completed in the next 45 min.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Biological cybernetics 38 (1980), S. 213-221 
    ISSN: 1432-0770
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Computer Science , Physics
    Notes: Abstract Identification of figural elements is based upon comparison of one of several formal pattern descriptions with subjective similarity judgements. The best description appears to contain figural elements which are heavily dependent on the visual domain chosen for the experiment. Generally those figural elements seem to be involved which distinguish optimally between the reference patterns used to mark off the visual domain. An algorithm is proposed for searching figural elements which are building stones of optimal descriptions. The present study indicates that grammars generating structures based upon features, higher order figures and rules of composition may be tools for visual research.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1420-9071
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary It is demonstrated that systolic and diastolic depolarization rate are correlated with the percentage of myofilaments in the cells of the rabbit sinoatrial node. It appears that, in the rabbit sinoatrial node, little-excitable transitional cells exist in the zone of propagation at the septal side of the typical nodal cells.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-2013
    Keywords: Aggregates ; Neonatal rat heart ; Slow inward current ; Inactivation ; Voltage clamp ; D600
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract In voltage clamped neonatal rat heart cells a transient current is observed during depolarizing potential steps, which was identified as slow inward current (I si) by its range of activation, by its reversal potential of approximately +50 mV and by its sensitivity to D600 or low external Ca2+. ThisI si activates too fast to be detected by the present methods, which implies that activation is completed within milliseconds. The time constant of inactivation was weakly potential dependent and less than 30 ms between −40 mV and +20 mV. Thef ∞ curve ofI si had a sigmoidal shape with 90% and 10% values near −50 mV and −10 mV respectively, half maximum was at −25 mV. From double pulse experiments an estimate was obtained of the potential dependence and amplitude of steady stateI si. A maximum was expected around −30 mV. Steady stateI si appears to be present indeed in steady state current voltage relations, as the relative minimum at −30 mV in such relations is abolished by 5·10−7 g/ml D600. Currents tails during hyperpolarizing steps from prepulse potentials near 0 mV are potential dependent in a way expected whenI si contributes to these current tails by a decrease in inactivation. Moreover, the current tails are diminished by D600 or Co2+. Consequenses of steady stateI si are discussed.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Pflügers Archiv 414 (1989), S. 95-98 
    ISSN: 1432-2013
    Keywords: cardiac fibroblasts and myocytes ; voltage clamp ; gating ; gap junction channel conductance ; connexons
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Recently, the use of the double whole-cell patchclamp technique enable conductance measurements of single gap junctional channels. Different values have been measured in pairs of rat lacrimal cells (6), murine acinar cells and chinese hamster ovary cells (9), embryonic chick heart- (10) and neonatal rat heart myocytes (7). We here present evidence that the conductance of gap junction channels between two different cell types originating from the same tissue, neonatal rat heart, is different. In mixed cultures of cardiac fibroblasts and myocytes, gap junction channels between fibroblasts have a single channel conductance of only 22 pS, while those between myocytes have a conductance of 43 pS. Fibroblasts can be electrically coupled to myocytes through channels having an intermediate conductance of 29 pS, a value which matches very well with te theoretically expected conductance of a gap junction channel composed of a fibroblast- and a myoblast connexon (hemichannel). These data provide direct evidence on the single channel level that in heterologous gap junction channels the composing connexons retain their cell-specific properties.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1432-2013
    Keywords: cardiac myocytes ; voltage clamp ; perforated patch method ; connexin43 ; guanosine 3′,5′-cyclic monophosphate ; carbachol ; alkaline phosphatase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The effects of an increase in intracellular cyclic GMP (cGMP)-concentration on gap junctional current (Ij) were studied in cultured neonatal rat heart cells using both the whole-cell and perforated patch voltage-clamp method. In whole-cell measurements, exposure to 8-bromo-cGMP or carbachol reduced Ij. With the perforated patch technique, on the other hand, Ij was not affected by either 8-bromo-cGMP or carbachol. Addition of alkaline phosphatase prevented the carbachol-induced decrease in Ij in whole-cell measurements. Reduction of Ij in well-coupled cell pairs by application of heptanol allowed us to study the effects of these substances on the single gap junction channel level. We found that cGMP-treatment shifts the single channel conductance (γj) from 43 to 21 pS in whole-cell measurements and that intracellular addition of phosphatase prevents this shift. In contrast, intracellular phosphatase-treatment itself shifts γj to 70 pS. Our results indicate that Cx43-gap junction channels may exhibit three conductance levels (21 pS, 40–45 pS and 70 pS), depending on the phosphorylation state of the protein.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Pflügers Archiv 431 (1995), S. 138-140 
    ISSN: 1432-2013
    Keywords: Connexins ; Voltage-clamp ; Intracellular pH
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Intercellular communication through gap junction channels can be regulated by changes in intracellular pH (pHi). This regulation may play an important role in ischemic heart tissue. Using the dual voltage-clamp technique, we compared the pHi sensitivity of gap junction channels composed of connexin 43 (Cx43) and Cx45, two of the gap junction proteins that are expressed in heart. We made use of SKHep1 cells, endogenously expressing low levels of Cx45 and SKHep1 cells stably transfected with rat Cx43. To manipulate the pHi we applied the NH3/NH4 + pH-clamp method. At pHi 6.7 the gj of Cx45 channels was reduced to ∼20% of control values (pHi 7.0) and at pHi 6.3 all channels closed. The gj of Cx43 channels was ∼70% of control values at pHi 6.7 and ∼40% at pHi 6.3. Cx43 channels closed at pHi 5.8. Single channel conductances were 17.8 pS for Cx45 and 40.8 pS for Cx43 at pHi 7.0 and did not change significantly at lower pHi. This suggests that the decrease in macroscopic conductance observed at low pHi results from the decrease in open probability of gap junctional channels rather than from a decrease in single channel conductance. Our results demonstrate that gap junction channels built of Cx45 are far more pH sensitive than channels built of Cx43.
    Type of Medium: Electronic Resource
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Basic research in cardiology 82 (1987), S. 454-464 
    ISSN: 1435-1803
    Keywords: cultured rat-heart cells ; beat-ratesynchronization ; gap junctions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary When two spontaneously beating neonatal rat heart cells in tissue culture were allowed to grow together they synchronized their originally independent beats to a common rhythm, as measured with an opto-electronic technique. Both single isolated cells and cell pairs exhibited a highly irregular beating pattern. Beating irregularity was strongly and positively correlated with mean interbeat interval. Synchronization of beating occurred in 50% of the pairs studied within one beating interval. In the remaining cell pairs, the first synchronized beat was followed by a 4–65 s period of partial synchronization. The time difference between contraction moments of two cells in a pair respective to each other (latency) changed upon synchronization from a random value to a fixed value. In a few cases the latency decreased during 20 to 30 s after the first synchronized beat before a steady-state value was reached. The mean interbeat interval (IBI) of the synchronized cell pairs was governed by the mean IBI of the originally faster beating cel. In 83% of the cases the mean IBI of the cell pairs was between that of the originally isolated beating cells. We conclude from the experiments described that physical coupling (i.e. gap junction formation) is virtually complete before beating synchronization occurs.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1573-2657
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The relationship between the calcium concentration and the isometric tension obtained with different techniques of skinning provides information on the biochemical events of contraction in vascular smooth muscle. Muscle preparations of the rabbit femoral artery were skinned with triton X-100, saponin, β-escin and α-toxin and the relationship between the calcium concentration and isometric tension was determined at different preparation lengths. We determined the calcium sensitivity as a function of muscle length with different techniques of skinning. At a pCa of 6.0, triton X-100 skinned smooth muscle of the femoral artery generated 50% of the maximal tension. In α-toxin skinned preparations, this calcium sensitivity was shifted to a pCa of 5.6. The sensitivity of the saponin and β-escin skinned preparations were in between those of the triton X-100 and the α-toxin skinned preparations. The cooperativity of the regulation of contraction varied among the differently skinned preparations between 3 (α-toxin) and 6 (triton X-100). The relationships between the calcium concentration and the isometric tension of the differently skinned preparations up to the optimal length for tension generation did not exhibit any length dependency. The length tension relationship, obtained from the maximal response at the highest calcium concentration is in line with that from other studies. The presence of intracellular proteins and membranes affects the regulation of contraction in smooth muscle of the femoral artery.
    Type of Medium: Electronic Resource
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