Length vs. active force relationship in single isolated smooth muscle cells

1991 ◽  
Vol 260 (5) ◽  
pp. C1104-C1112 ◽  
Author(s):  
D. E. Harris ◽  
D. M. Warshaw
Keyword(s):  
Smooth Muscle ◽  
Elastic Modulus ◽  
Smooth Muscle Cells ◽  
Force Measurement ◽  
Muscle Length ◽  
Muscle Cells ◽  
Active Force ◽  
Length Changes ◽  
Cross Bridges ◽  
Force Relationship

The length vs. active force relationship (L-F) may provide information about changes in smooth muscle contractile protein interactions as muscle length changes. To characterize the L-F in single toad stomach smooth muscle cells, cells were attached to a force measurement system, electrically stimulated, and isometric force and elastic modulus (an estimate of the number of attached cross bridges) determined at different cell lengths. Cells generated maximum stress (Pmax = 152.5 mN/mm2) and elastic modulus (Eact = 0.68 x 10(4) mN/mm2) at their rest length (Lcell = 78.0 microns; distance between cell attachments). At shorter lengths, active force and elastic modulus declined proportionally with active force eliminated at 0.4 Lcell. Stretching the relaxed cells up to 1.4 Lcell shifted the subsequent L-F along the length axis by the amount of the stretch but did not change Pmax or the shape of the L-F. In activated cells, force was a function of cell length rather than of shortening history. We interpret these findings as evidence that 1) Lcell is close to the optimum length for force generation, 2) the decline in force at lengths less than Lcell results from a reduced number of attached cross bridges, and 3) stretching relaxed smooth muscle cells may not move the contractile units to new positions on their L-F.

Agonist activation modulates cross-bridge states in single vascular smooth muscle cells

1993 ◽  
Vol 264 (1) ◽  
pp. C103-C108 ◽  
Author(s):  
F. V. Brozovich ◽  
M. Yamakawa
Keyword(s):  
Smooth Muscle ◽  
Steady State ◽  
Vascular Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Vascular Smooth Muscle Cells ◽  
Muscle Cells ◽  
Relative Population ◽  
Cross Bridge ◽  
Cross Bridges ◽  
State Force

To determine cross-bridge properties during agonist-stimulated contractions, steady-state force and relative steady-state stiffness were recorded at rest (pCa 9) and during both full (pCa 4) and partial (pCa 7) Ca2+ activations of isolated single alpha-toxin permeabilized vascular smooth muscle cells. For pCa 4 and pCa 7, agonist (1 microM histamine) activation resulted in significant (P < 0.05) increases in both force and stiffness. The agonist-induced increase of steady-state force was significantly (P < 0.05) greater than that of stiffness; at pCa 4, there was a 48% increase for force vs. 17% for stiffness, and, at pCa 7, there was a 160% increase for force vs. 57% for stiffness. The increase in force and stiffness after agonist prestimulation implies that the number of attached cross bridges has increased. However, after agonist prestimulation, we found that the increase of force was greater (P < 0.05) than that of stiffness, resulting in a greater force at any given level of stiffness. Thus these data indicate that agonist activation, presumably via activation of a G protein, increases the relative force per attached cross bridge, possibly by modulating the kinetics of the actomyosin adenosinetriphosphatase to increase in the relative population of cross bridges in force-producing states [actinomyosin (AM) or AM.ADP].

scholarly journals Force: velocity relationship in single isolated toad stomach smooth muscle cells.

1987 ◽  
Vol 89 (5) ◽  
pp. 771-789 ◽  
Author(s):  
D M Warshaw
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Striated Muscle ◽  
Muscle Cells ◽  
Length Change ◽  
Force Transducer ◽  
Cell Length ◽  
Shortening Velocity ◽  
Cross Sectional ◽  
Cross Bridges

The relationship between force and shortening velocity (F:V) in muscle is believed to reflect both the mechanics of the myosin cross-bridge and the kinetics of its interaction with actin. To date, the F:V for smooth muscle cells has been inferred from F:V data obtained in multicellular tissue preparations. Therefore, to determine F:V in an intact single smooth muscle cell, cells were isolated from the toad (Bufo marinus) stomach muscularis and attached to a force transducer and length displacement device. Cells were electrically stimulated at 20 degrees C and generated 143 mN/mm2 of active force per muscle cross-sectional area. At the peak of contraction, cells were subjected to sudden changes in force (dF = 0.10-0.90 Fmax) and then maintained at the new force level. The force change resulted in a length response in which the cell length (Lcell) rapidly decreased during the force step and then decreased monotonically with a time constant between 75 and 600 ms. The initial length change that coincided with the force step was analyzed and an active cellular compliance of 1.9% cell length was estimated. The maintained force and resultant shortening velocity (V) were fitted to the Hill hyperbola with constants a/Fmax of 0.268 and b of 0.163 Lcell/s. Vmax was also determined by a procedure in which the cell length was slackened and the time of unloaded shortening was recorded (slack test). From the slack test, Vmax was estimated as 0.583 Lcell/s, in agreement with the F:V data. The F:V data were analyzed within the framework of the Huxley model (Huxley. 1957. Progress in Biophysics and Biophysical Chemistry. 7:255-318) for contraction and interpreted to indicate that in smooth muscle, as compared with fast striated muscle, there may exist a greater percentage of attached force-generating cross-bridges.

scholarly journals Involvement of gap junctions between smooth muscle cells in sustained hypoxic pulmonary vasoconstriction development: a potential role for 15-HETE and 20-HETE

2016 ◽  
Vol 310 (8) ◽  
pp. L772-L783 ◽  
Author(s):  
Igor V. Kizub ◽  
Anand Lakhkar ◽  
Vidhi Dhagia ◽  
Sachindra R. Joshi ◽  
Houli Jiang ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Gap Junctions ◽  
Force Measurement ◽  
Isometric Force ◽  
Hypoxic Pulmonary Vasoconstriction ◽  
Muscle Cells ◽  
Cytochrome P450 Monooxygenases ◽  
Prostaglandin E ◽  
Pulmonary Vasoconstriction

In response to hypoxia, the pulmonary artery normally constricts to maintain optimal ventilation-perfusion matching in the lung, but chronic hypoxia leads to the development of pulmonary hypertension. The mechanisms of sustained hypoxic pulmonary vasoconstriction (HPV) remain unclear. The aim of this study was to determine the role of gap junctions (GJs) between smooth muscle cells (SMCs) in the sustained HPV development and involvement of arachidonic acid (AA) metabolites in GJ-mediated signaling. Vascular tone was measured in bovine intrapulmonary arteries (BIPAs) using isometric force measurement technique. Expression of contractile proteins was determined by Western blot. AA metabolites in the bath fluid were analyzed by mass spectrometry. Prolonged hypoxia elicited endothelium-independent sustained HPV in BIPAs. Inhibition of GJs by 18β-glycyrrhetinic acid (18β-GA) and heptanol, nonspecific blockers, and Gap-27, a specific blocker, decreased HPV in deendothelized BIPAs. The sustained HPV was not dependent on Ca2+entry but decreased by removal of Ca2+and by Rho-kinase inhibition with Y-27632. Furthermore, inhibition of GJs decreased smooth muscle myosin heavy chain (SM-MHC) expression and myosin light chain phosphorylation in BIPAs. Interestingly, inhibition of 15- and 20-hydroxyeicosatetraenoic acid (HETE) synthesis decreased HPV in deendothelized BIPAs. 15-HETE- and 20-HETE-stimulated constriction of BIPAs was inhibited by 18β-GA and Gap-27. Application of 15-HETE and 20-HETE to BIPAs increased SM-MHC expression, which was also suppressed by 18β-GA and by inhibitors of lipoxygenase and cytochrome P450 monooxygenases. More interestingly, 15,20-dihydroxyeicosatetraenoic acid and 20-OH-prostaglandin E2, novel derivatives of 20-HETE, were detected in tissue bath fluid and synthesis of these derivatives was almost completely abolished by 18β-GA. Taken together, our novel findings show that GJs between SMCs are involved in the sustained HPV in BIPAs, and 15-HETE and 20-HETE, through GJs, appear to mediate SM-MHC expression and contribute to the sustained HPV development.

scholarly journals Modeling the impairment of airway smooth muscle force by stretch

2015 ◽  
Vol 118 (6) ◽  
pp. 684-691 ◽  
Author(s):  
Jason H. T. Bates
Keyword(s):  
Smooth Muscle ◽  
Airway Smooth Muscle ◽  
Coarse Grained ◽  
Active Force ◽  
Cross Bridge ◽  
Bulk Force ◽  
Length Changes ◽  
Cross Bridges ◽  
Large Stretch ◽  
Two State Model

Imposed length changes of only a small percent produce transient reductions in active force in strips of airway smooth muscle (ASM) due to the temporary detachment of bound cross-bridges caused by the relative motion of the actin and myosin fibers. More dramatic and sustained reductions in active force occur following large changes in length. The Huxley two-state model of skeletal muscle originally proposed in 1957 and later adapted to include a four-state description of cross-bridge kinetics has been widely used to model the former phenomenon, but is unable to account for the latter unless modified to include mechanisms by which the contractile machinery in the ASM cell becomes appropriately rearranged. Even so, the Huxley model itself is based on the assumption that the contractile proteins are all aligned precisely in the direction of bulk force generation, which is not true for ASM. The present study derives a coarse-grained version of the Huxley model that is free of inherent assumptions about cross-bridge orientation. This simplified model recapitulates the key features observed in the force-length behavior of activated strips of ASM and, in addition, provides a mechanistically based way of accounting for the sustained force reductions that occur following large stretch.

scholarly journals Matrix stiffness-modulated proliferation and secretory function of the airway smooth muscle cells

2015 ◽  
Vol 308 (11) ◽  
pp. L1125-L1135 ◽  
Author(s):  
Artem Shkumatov ◽  
Michael Thompson ◽  
Kyoung M. Choi ◽  
Delphine Sicard ◽  
Kwanghyun Baek ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Smooth Muscle ◽  
Elastic Modulus ◽  
Smooth Muscle Cells ◽  
Airway Smooth Muscle ◽  
Tissue Remodeling ◽  
Muscle Cells ◽  
Bioactive Molecules ◽  
Airway Smooth Muscle Cells ◽  
Mechanical Stiffness

Multiple pulmonary conditions are characterized by an abnormal misbalance between various tissue components, for example, an increase in the fibrous connective tissue and loss/increase in extracellular matrix proteins (ECM). Such tissue remodeling may adversely impact physiological function of airway smooth muscle cells (ASMCs) responsible for contraction of airways and release of a variety of bioactive molecules. However, few efforts have been made to understand the potentially significant impact of tissue remodeling on ASMCs. Therefore, this study reports how ASMCs respond to a change in mechanical stiffness of a matrix, to which ASMCs adhere because mechanical stiffness of the remodeled airways is often different from the physiological stiffness. Accordingly, using atomic force microscopy (AFM) measurements, we found that the elastic modulus of the mouse bronchus has an arithmetic mean of 23.1 ± 14 kPa (SD) (median 18.6 kPa). By culturing ASMCs on collagen-conjugated polyacrylamide hydrogels with controlled elastic moduli, we found that gels designed to be softer than average airway tissue significantly increased cellular secretion of vascular endothelial growth factor (VEGF). Conversely, gels stiffer than average airways stimulated cell proliferation, while reducing VEGF secretion and agonist-induced calcium responses of ASMCs. These dependencies of cellular activities on elastic modulus of the gel were correlated with changes in the expression of integrin-β1 and integrin-linked kinase (ILK). Overall, the results of this study demonstrate that changes in matrix mechanics alter cell proliferation, calcium signaling, and proangiogenic functions in ASMCs.

Adjustable passive length-tension curve in rabbit detrusor smooth muscle

2007 ◽  
Vol 102 (5) ◽  
pp. 1746-1755 ◽  
Author(s):  
John E. Speich ◽  
Christopher Dosier ◽  
Lindsey Borgsmiller ◽  
Kevin Quintero ◽  
Harry P. Koo ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Strain Softening ◽  
Muscle Length ◽  
Strain History ◽  
Total Force ◽  
Detrusor Smooth Muscle ◽  
Passive Force ◽  
Passive Stiffness ◽  
Active Force ◽  
Length Changes

Until the 1990s, the passive and active length-tension ( L-T) relationships of smooth muscle were believed to be static, with a single passive force value and a single maximum active force value for each muscle length. However, recent studies have demonstrated that the active L-T relationship in airway smooth muscle is dynamic and adapts to length changes over a period of time. Furthermore, our prior work showed that the passive L-T relationship in rabbit detrusor smooth muscle (DSM) is also dynamic and that in addition to viscoelastic behavior, DSM displays strain-softening behavior characterized by a loss of passive stiffness at shorter lengths following a stretch to a new longer length. This loss of passive stiffness appears to be irreversible when the muscle is not producing active force and during submaximal activation but is reversible on full muscle activation, which indicates that the stiffness component of passive force lost to strain softening is adjustable in DSM. The present study demonstrates that the passive L-T curve for DSM is not static and can shift along the length axis as a function of strain history and activation history. This study also demonstrates that adjustable passive stiffness (APS) can modulate total force (35% increase) for a given muscle length, while active force remains relatively unchanged (4% increase). This finding suggests that the structures responsible for APS act in parallel with the contractile apparatus, and the results are used to further justify the configuration of modeling elements within our previously proposed mechanical model for APS.

Synergistic actions of insulin and troglitazone on contractility in endothelium-denuded rat aortic rings

1998 ◽  
Vol 275 (5) ◽  
pp. E882-E887 ◽  
Author(s):  
Chetan Goud ◽  
Bertram Pitt ◽  
R. Clinton Webb ◽  
Joyce M. Richey
Keyword(s):  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Vascular Smooth Muscle Cells ◽  
Vascular Reactivity ◽  
Force Measurement ◽  
Muscle Cells ◽  
Bay K 8644 ◽  
Vascular Contraction ◽  
Intracellular Ca

Insulin attenuates vascular contraction via inhibition of voltage-operated Ca2+ channels and by enhancement of endothelium-dependent vasodilation. Thus it has been suggested that hypertension-associated insulin resistance results from an insensitivity to the hormone’s effects on vascular reactivity. This hypothesis has been strengthened by reports that thiazolidinediones, a class of insulin-sensitizing agents, lower blood pressure and improve insulin responsiveness in hypertensive, insulin-resistant animal models. We tested the hypothesis that troglitazone enhances the vasodilating effect of insulin via inhibition of voltage-operated Ca2+ channels in vascular smooth muscle cells. Rat thoracic aortic rings (no endothelium) were suspended in tissue baths for isometric force measurement. Rings were incubated with 0.1 DMSO vehicle (control), troglitazone (10−5 M), insulin (10−7 U/l), or both troglitazone and insulin (1 h) and then contracted with phenylephrine (PE), KCl, or BAY K 8644. Troglitazone increased the EC50 values for PE and KCl. Contractions to BAY K 8644 in troglitazone-treated rings were virtually abolished. Insulin alone had no effect on contraction. However, when insulin was combined with troglitazone, the EC50 values for PE and KCl were further increased. Additionally, the maximum contractions to both PE (14 ± 4% of control) and KCl (12 ± 2% of control) were reduced. Measurement of Ca2+concentration ([Ca2+]) with fura 2-AM in dispersed vascular smooth muscle cells indicated that neither insulin nor troglitazone alone altered PE-induced increases in intracellular [Ca2+]. However, troglitazone and insulin together caused a significant reduction in PE-induced increases in intracellular [Ca2+] (expressed as percentage of preincubation stimulation to PE: 47 ± 10%, treated; 102 ± 13%, vehicle). These results demonstrate that troglitazone inhibits Ca2+ influx and that it acts synergistically with insulin to attenuate further vascular contraction via inhibition of voltage-operated Ca2+ channels.

Immunogold localization of HMB-45 antigen in pulmonary lymphangiomyomatosis

1995 ◽  
Vol 53 ◽  
pp. 998-999
Author(s):  
J.M. Minda ◽  
E. Dessy ◽  
G. G. Pietra
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Room Temperature ◽  
Osmium Tetroxide ◽  
Muscle Cells ◽  
Ultrastructural Localization ◽  
Reproductive Age ◽  
Thin Sections ◽  
Smooth Muscle Proliferation ◽  
Hmb 45

Pulmonary lymphangiomyomatosis (PLAM) is a rare disease occurring exclusively in women of reproductive age. It involves the lungs, lymph nodes and lymphatic ducts. In the lungs, it is characterized by the proliferation of smooth muscle cells around lymphatics in the bronchovascular bundles, lobular septa and pleura The nature of smooth muscle proliferation in PLAM is still unclear. Recently, reactivity of the smooth muscle cells for HMB-45, a melanoma-related antigen has been reported by immunohistochemistry. The purpose of this study was the ultrastructural localization of HMB-45 immunoreactivity in these cells using gold-labeled antibodies.Lung tissue from three cases of PLAM, referred to our Institution for lung transplantation, was embedded in either Poly/Bed 812 post-fixed in 1% osmium tetroxide, or in LR White, without osmication. For the immunogold technique, thin sections were placed on Nickel grids and incubated with affinity purified, monoclonal anti-melanoma antibody HMB-45 (1:1) (Enzo Diag. Co) overnight at 4°C. After extensive washing with PBS, grids were treated with Goat-anti-mouse-IgG-Gold (5nm) (1:10) (Amersham Life Sci) for 1 hour, at room temperature.

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