Dependence of fluid-phase pinocytosis in arterial smooth-muscle cells on temperature, cellular ATP concentration and the cytoskeletal system.
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ABSTRACT:
SUBMITTER: Muir EM
PROVIDER: S-EPMC1152525 | biostudies-other | 1983 Nov
REPOSITORIES: biostudies-other
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Project description:Arterial smooth muscle (SM) cells respond autonomously to changes in intravascular pressure, adjusting tension to maintain vessel diameter. The values of membrane potential (Vm) and sarcoplasmic Ca(2+) concentration (Ca(in)) within minutes of a change in pressure are the results of two opposing pathways, both of which use Ca(2+) as a signal. This works because the two Ca(2+)-signaling pathways are confined to distinct microdomains in which the Ca(2+) concentrations needed to activate key channels are transiently higher than Ca(in). A mathematical model of an isolated arterial SM cell is presented that incorporates the two types of microdomains. The first type consists of junctions between cisternae of the peripheral sarcoplasmic reticulum (SR), containing ryanodine receptors (RyRs), and the sarcolemma, containing voltage- and Ca(2+)-activated K(+) (BK) channels. These junctional microdomains promote hyperpolarization, reduced Ca(in), and relaxation. The second type is postulated to form around stretch-activated nonspecific cation channels and neighboring Ca(2+)-activated Cl(-) channels, and promotes the opposite (depolarization, increased Ca(in), and contraction). The model includes three additional compartments: the sarcoplasm, the central SR lumen, and the peripheral SR lumen. It incorporates 37 protein components. In addition to pressure, the model accommodates inputs of ?- and ?-adrenergic agonists, ATP, 11,12-epoxyeicosatrienoic acid, and nitric oxide (NO). The parameters of the equations were adjusted to obtain a close fit to reported Vm and Ca(in) as functions of pressure, which have been determined in cerebral arteries. The simulations were insensitive to ± 10% changes in most of the parameters. The model also simulated the effects of inhibiting RyR, BK, or voltage-activated Ca(2+) channels on Vm and Ca(in). Deletion of BK ?1 subunits is known to increase arterial-SM tension. In the model, deletion of ?1 raised Ca(in) at all pressures, and these increases were reversed by NO.
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Project description:During isometric activation of airway smooth muscle (ASM), cross-bridge cycling and ATP hydrolysis rates decline across time even though isometric force is sustained. Thus, tension cost (i.e., ATP hydrolysis rate per unit of force during activation) decreases with time. The "latch-state" hypothesis attributes the dynamic change in cross-bridge cycling and ATP hydrolysis rates to changes in phosphorylation of the regulatory myosin light chain (rMLC20 ). However, we previously showed that in ASM, the extent of rMLC20 phosphorylation remains unchanged during sustained isometric force. As an alternative, we hypothesized that cytoskeletal remodeling within ASM cells results in increased internal loading of contractile proteins that slows cross-bridge cycling and ATP hydrolysis rates. To test this hypothesis, we simultaneously measured isometric force and ATP hydrolysis rate in permeabilized porcine ASM strips activated by Ca2+ (pCa 4.0). The extent of rMLC20 phosphorylation remained unchanged during isometric activation, even though ATP hydrolysis rate (tension cost) declined with time. The effect of cytoskeletal remodeling was assessed by inhibiting actin polymerization using Cytochalasin D (Cyto-D). In Cyto-D treated ASM, isometric force was reduced while ATP hydrolysis rate increased compared to untreated ASM strips. These results indicate that external transmission of force, cross-bridge cycling and ATP hydrolysis rates are affected by internal loading of contractile proteins.
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