Nitrovasodilators relax arterial smooth muscle by decreasing [Ca2+]i and uncoupling stress from myosin phosphorylation

1992 ◽  
Vol 263 (2) ◽  
pp. C461-C467 ◽  
Author(s):  
N. L. McDaniel ◽  
X. L. Chen ◽  
H. A. Singer ◽  
R. A. Murphy ◽  
C. M. Rembold
Keyword(s):  
Smooth Muscle ◽  
Myosin Light Chain ◽  
Carotid Arteries ◽  
Muscle Relaxation ◽  
Smooth Muscle Relaxation ◽  
Myosin Light Chain Phosphorylation ◽  
Arterial Smooth Muscle ◽  
Histamine Stimulation ◽  
Myosin Phosphorylation ◽  
Proportional Decrease

Elevations in guanosine 3',5'-cyclic monophosphate concentration ([cGMP]) are proposed to induce arterial smooth muscle relaxation by either 1) decreasing myoplasmic [Ca2+] ([Ca2+]i), 2) decreasing the [Ca2+]i sensitivity of phosphorylation, or 3) uncoupling force from myosin phosphorylation. We evaluated the importance of each of these mechanisms by measuring changes in [cGMP], aequorin- and fura-2-estimated [Ca2+]i, myosin light chain phosphorylation, and stress in histamine-stimulated swine carotid arteries. In tissues submaximally stimulated with 3 microM histamine, nitroprusside (NP) induced a proportional decrease in myoplasmic [Ca2+] and myosin phosphorylation, suggesting that the relaxation was at least partially induced by decreases in [Ca2+]i without a change in the [Ca2+]i sensitivity of phosphorylation. In tissues maximally stimulated with 10 microM histamine, NP and nitroglycerin produced significant relaxations that were not associated with significant sustained reductions in [Ca2+]i or myosin phosphorylation. With both submaximal and maximal histamine stimulation, nitrovasodilators produced more substantial relaxation than that expected from the nitrovasodilator-induced reduction in myosin phosphorylation. These results suggest that nitrovasodilators relax histamine-stimulated swine arterial smooth muscle by at least two mechanisms: 1) reducing [Ca2+]i, an effect observed in submaximally stimulated tissues, and 2) uncoupling of stress from myosin phosphorylation.

scholarly journals Arteriolar vasodilation involves actin depolymerization

2018 ◽  
Vol 315 (2) ◽  
pp. H423-H428
Author(s):  
Philip S. Clifford ◽  
Brian S. Ferguson ◽  
Jeffrey L. Jasperse ◽  
Michael A. Hill
Keyword(s):  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Sodium Nitroprusside ◽  
Light Chain ◽  
Myosin Light Chain ◽  
Muscle Relaxation ◽  
Nitric Oxide Donor ◽  
Smooth Muscle Relaxation ◽  
Myosin Light Chain Phosphorylation ◽  
Actin Depolymerization

It is generally assumed that relaxation of arteriolar vascular smooth muscle occurs through hyperpolarization of the cell membrane, reduction in intracellular Ca2+ concentration, and activation of myosin light chain phosphatase/inactivation of myosin light chain kinase. We hypothesized that vasodilation is related to depolymerization of F-actin. Cremaster muscles were dissected in rats under pentobarbital sodium anesthesia (50 mg/kg). First-order arterioles were dissected, cannulated on glass micropipettes, pressurized, and warmed to 34°C. Internal diameter was monitored with an electronic video caliper. The concentration of G-actin was determined in flash-frozen intact segments of arterioles by ultracentrifugation and Western blot analyses. Arterioles dilated by ~40% of initial diameter in response to pinacidil (1 × 10−6 mM) and sodium nitroprusside (5 × 10−5 mM). The G-actin-to-smooth muscle 22α ratio was 0.67 ± 0.09 in arterioles with myogenic tone and increased significantly to 1.32 ± 0.34 ( P < 0.01) when arterioles were dilated with pinacidil and 1.14 ± 0.18 ( P < 0.01) with sodium nitroprusside, indicating actin depolymerization. Compared with control vessels (49 ± 5%), the percentage of phosphorylated myosin light chain was significantly reduced by pinacidil (24 ± 2%, P < 0.01) but not sodium nitroprusside (42 ± 4%). These findings suggest that actin depolymerization is an important mechanism for vasodilation of resistance arterioles to external agonists. Furthermore, pinacidil produces smooth muscle relaxation via both decreases in myosin light chain phosphorylation and actin depolymerization, whereas sodium nitroprusside produces smooth muscle relaxation primarily via actin depolymerization. NEW & NOTEWORTHY This article adds to the accumulating evidence on the contribution of the actin cytoskeleton to the regulation of vascular smooth muscle tone in resistance arterioles. Actin depolymerization appears to be an important mechanism for vasodilation of resistance arterioles to pharmacological agonists. Dilation to the K+ channel opener pinacidil is produced by decreases in myosin light chain phosphorylation and actin depolymerization, whereas dilation to the nitric oxide donor sodium nitroprusside occurs primarily via actin depolymerization. Listen to this article’s corresponding podcast at https://ajpheart.podbean.com/e/vascular-smooth-muscle-actin-depolymerization/ .

Cyclic nucleotide-dependent regulation of Mn2+ influx, [Ca2+]i, and arterial smooth muscle relaxation

1992 ◽  
Vol 263 (2) ◽  
pp. C468-C473 ◽  
Author(s):  
X. L. Chen ◽  
C. M. Rembold
Keyword(s):  
Smooth Muscle ◽  
Cyclic Nucleotides ◽  
Cyclic Nucleotide ◽  
Channel Blocker ◽  
Muscle Relaxation ◽  
Contractile Force ◽  
Smooth Muscle Relaxation ◽  
Quenching Rate ◽  
Arterial Smooth Muscle ◽  
Histamine Stimulation

Elevations in cyclic nucleotide levels can decrease myoplasmic [Ca2+] ([Ca2+]i) and thereby induce arterial smooth muscle relaxation. We evaluated whether cyclic nucleotide-induced reductions in [Ca2+]i are caused by 1) decreased Ca2+ influx or 2) increased Ca2+ sequestration or efflux. Swine carotid medial tissues were loaded with fura-2, and Ca2+ influx was estimated from the quenching rate of 360-nm fluorescence after addition of extracellular Mn2+. Histamine stimulation or high KCl depolarization increased Mn2+ influx, [Ca2+]i, and contractile force. The Ca2+ channel blocker diltiazem attenuated histamine- or KCl-induced increases in Mn2+ influx, [Ca2+]i, and force. Addition of forskolin (which increases cAMP) or nitroglycerin (which increases cGMP) attenuated histamine-induced increases in Mn2+ influx, [Ca2+]i, and force. Addition of forskolin or nitroglycerin also relaxed KCl depolarized tissues; however, Mn2+ influx and [Ca2+]i remained high. These results suggest that Mn(2+)-induced quenching of 360-nm fluorescence is an estimate of Ca2+ influx in the intact swine carotid artery. These results also suggest that cyclic nucleotides can relax swine arterial smooth muscle by at least two mechanisms: 1) reduction of [Ca2+]i primarily induced by decreases in Ca2+ influx and 2) uncoupling force from [Ca2+]i without changing Ca2+ influx or [Ca2+]i.

Telokin expression and the effect of hypoxia on its phosphorylation status in smooth muscle cells from small and large pulmonary arteries

2008 ◽  
Vol 294 (6) ◽  
pp. L1166-L1173 ◽  
Author(s):  
Jane A. Madden ◽  
Mark W. Dantuma ◽  
Elena A. Sorokina ◽  
Dorothee Weihrauch ◽  
Jack G. Kleinman
Keyword(s):  
Smooth Muscle ◽  
Muscle Cell ◽  
Light Chain ◽  
Myosin Light Chain ◽  
Muscle Relaxation ◽  
Cerebral Arteries ◽  
Pulmonary Arteries ◽  
Smooth Muscle Relaxation ◽  
Myosin Phosphatase ◽  
Mlc Phosphorylation

Small pulmonary arteries (SPA), <500 μm diameter of the cat, constrict when exposed to hypoxia, whereas larger arteries (large pulmonary arteries; LPA), >800 μm diameter, show little or no response. It is unknown why different contractile responses occur within the same vascular bed, but activator or repressor proteins within the smooth muscle cell (SMC) can modify myosin phosphatase and myosin light chain kinase (MLCK), thereby influencing the phosphorylation state of myosin light chain (MLC) and ultimately, contraction. Telokin, a protein with a sequence identical to the COOH-terminal domain of MLCK, is expressed in smooth muscle where in its phosphorylated state it inhibits myosin phosphatase, binds to unphosphorylated myosin, and helps maintain smooth muscle relaxation. We measured telokin mRNA and telokin protein in smooth muscle from different diameter feline pulmonary arteries and sought to determine whether changes in the phosphorylation status of telokin and MLC occurred during hypoxia. In pulmonary arteries, telokin expression varied inversely with artery diameter, but cerebral arteries showed neither telokin protein nor telokin mRNA. Although telokin and MLC were distributed uniformly throughout the SPA muscle cell cytoplasm, they were not colocalized. During hypoxia, telokin dephosphorylated, and MLC became increasingly phosphorylated in SPA SMC, whereas in LPA SMC there was no change in either telokin or MLC phosphorylation. When LPA SMC were exposed to phenylephrine, MLC phosphorylation increased with no change in telokin phosphorylation. These results suggest that in SPA, phosphorylated telokin may help maintain relaxation under unstimulated conditions, whereas in LPA, telokin's function remains undetermined.

Effects of cGMP on [Ca2+]i, myosin light chain phosphorylation, and contraction in human myometrium

1991 ◽  
Vol 260 (4) ◽  
pp. C861-C867 ◽  
Author(s):  
R. A. Word ◽  
M. L. Casey ◽  
K. E. Kamm ◽  
J. T. Stull
Keyword(s):  
Smooth Muscle ◽  
Light Chain ◽  
Myosin Light Chain ◽  
Smooth Muscles ◽  
Smooth Muscle Relaxation ◽  
Human Myometrium ◽  
Myosin Light Chain Phosphorylation ◽  
Cyclic Monophosphate ◽  
Uterine Smooth Muscle ◽  
Spontaneous Contractions

Adenosine 3',5'-cyclic monophosphate (cAMP) is believed to be an important mediator of myometrial relaxation, and there is evidence to suggest that guanosine 3',5'-cyclic monophosphate (cGMP) is a mediator of smooth muscle relaxation in vascular and probably nonvascular tissues. To investigate the biochemical mechanisms involved in regulation of human myometrial contractility, we studied the effects of analogues of cAMP and cGMP, as well as activators of adenylate and guanylate cyclases, on uterine smooth muscle contractile activity. We found that myometrial smooth muscle cells in culture respond to analogues of cGMP and cAMP, as well as activators of guanylate cyclase, with a significant decrease in the resting and endothelin-induced increase in [Ca2+]i. Treatment of human uterine smooth muscle strips with sodium nitroprusside or isoproterenol results in diminished force and frequency of contraction as well as a decrease in the rate and extent of myosin light chain phosphorylation in spontaneous contractions of human myometrium. cGMP did not effect relaxation of endothelin-stimulated contractions of human myometrium, but the relaxation effects of cGMP were dramatic in precontracted bovine tracheal and human fetal aortic smooth muscles. Whereas cGMP and cAMP act to promote a decrease in the force and frequency of spontaneous contractions in human myometrium, this tissue is not as responsive to the actions of cyclic nucleotides as are other types of smooth muscle.

A myosin phosphatase targeting subunit isoform transition defines a smooth muscle developmental phenotypic switch

2000 ◽  
Vol 278 (3) ◽  
pp. C589-C600 ◽  
Author(s):  
Wessel P. Dirksen ◽  
Franjo Vladic ◽  
Steven A. Fisher
Keyword(s):  
Smooth Muscle ◽  
Alternative Splicing ◽  
Light Chain ◽  
Myosin Light Chain ◽  
Muscle Relaxation ◽  
Smooth Muscle Relaxation ◽  
Myosin Phosphatase ◽  
Mrna Isoforms ◽  
Regulatory Myosin Light Chain ◽  
Muscle Tissues

Smooth muscle myosin phosphatase dephosphorylates the regulatory myosin light chain and thus mediates smooth muscle relaxation. The activity of this myosin phosphatase is dependent upon its myosin-targeting subunit (MYPT1). Isoforms of MYPT1 have been identified, but how they are generated and their relationship to smooth muscle phenotypes is not clear. Cloning of the middle section of chicken and rat MYPT1 genes revealed that each gene gave rise to isoforms by cassette-type alternative splicing of exons. In chicken, a 123-nucleotide exon was included or excluded from the mature mRNA, whereas in rat two exons immediately downstream were alternative. MYPT1 isoforms lacking the alternative exon were only detected in mature chicken smooth muscle tissues that display phasic contractile properties, but the isoform ratios were variable. The patterns of expression of rat MYPT1 mRNA isoforms were more complex, with three major and two minor isoforms present in all smooth muscle tissues at varying stoichiometries. Isoform switching was identified in the developing chicken gizzard, in which the exon-skipped isoform replaced the exon-included isoform around the time of hatching. This isoform switch occurred after transitions in myosin heavy chain and myosin light chain (MLC17) isoforms and correlated with a severalfold increase in the rate of relaxation. The developmental switch of MYPT1 isoforms is a good model for determining the mechanisms and significance of alternative splicing in smooth muscle.

Sign in / Sign up

Export Citation Format

Share Document