93 research outputs found
Reverse mode Na+/Ca2+ exchange mediated by STIM1 contributes to Ca2+ influx in airway smooth muscle following agonist stimulation
<p>Abstract</p> <p>Background</p> <p>Agonist stimulation of airway smooth muscle (ASM) results in IP<sub>3 </sub>mediated Ca<sup>2+ </sup>release from the sarcoplasmic reticulum followed by the activation of store operated and receptor operated non-selective cation channels. Activation of these non-selective channels also results in a Na<sup>+ </sup>influx. This localised increase in Na<sup>+ </sup>levels can potentially switch the Na<sup>+</sup>/Ca<sup>2+ </sup>exchanger into reverse mode and so result in a further influx of Ca<sup>2+</sup>. The aim of this study was to characterise the expression and physiological function of the Na<sup>+</sup>/Ca<sup>2+ </sup>exchanger in cultured human bronchial smooth muscle cells and determine its contribution to agonist induced Ca<sup>2+ </sup>influx into these cells.</p> <p>Methods</p> <p>The expression profile of NCX (which encodes the Na<sup>+</sup>/Ca<sup>2+ </sup>exchanger) homologues in cultured human bronchial smooth muscle cells was determined by reverse transcriptase PCR. The functional activity of reverse mode NCX was investigated using a combination of whole cell patch clamp, intracellular Ca<sup>2+ </sup>measurements and porcine airway contractile analyses. KB-R7943 (an antagonist for reverse mode NCX) and target specific siRNA were utilised as tools to inhibit NCX function.</p> <p>Results</p> <p>NCX1 protein was detected in cultured human bronchial smooth muscle cells (HBSMC) cells and NCX1.3 was the only mRNA transcript variant detected. A combination of intracellular Na<sup>+ </sup>loading and addition of extracellular Ca<sup>2+ </sup>induced an outwardly rectifying current which was augmented following stimulation with histamine. This outwardly rectifying current was inhibited by 10 μM KB-R7943 (an antagonist of reverse mode NCX1) and was reduced in cells incubated with siRNA against NCX1. Interestingly, this outwardly rectifying current was also inhibited following knockdown of STIM1, suggesting for the first time a link between store operated cation entry and NCX1 activation. In addition, 10 μM KB-R7943 inhibited agonist induced changes in cytosolic Ca<sup>2+ </sup>and induced relaxation of porcine peripheral airways.</p> <p>Conclusions</p> <p>Taken together, these data demonstrate a potentially important role for NCX1 in control of Ca<sup>2+ </sup>homeostasis and link store depletion via STIM1 directly with NCX activation.</p
The Tetrodotoxin Binding Site Is within the Outer Vestibule of the Sodium Channel
Tetrodotoxin and saxitoxin are small, compact asymmetrical marine toxins that block voltage-gated Na channels with high affinity and specificity. They enter the channel pore’s outer vestibule and bind to multiple residues that control permeation. Radiolabeled toxins were key contributors to channel protein purification and subsequent cloning. They also helped identify critical structural elements called P loops. Spacial organization of their mutation-identified interaction sites in molecular models has generated a molecular image of the TTX binding site in the outer vestibule and the critical permeation and selectivity features of this region. One site in the channel’s domain I P loop determines affinity differences in mammalian isoforms
Tris(hydroxypropyl)phosphine Oxide: A Chiral Three-Dimensional Material with Nonlinear Optical Properties
Structural and electrical characterization of AuPtAlTi ohmic contacts to AlGaN/GaN with varying annealing temperature and Al content
The effect of varying annealing temperature and Al layer thickness on the structural and electrical characteristics of AuPtAlTi/AlGaN/GaN ohmic contact structures has been systematically investigated. The relationship between annealing temperature, Al content, interfacial microstructure, surface planarity and contact resistance is
examined. In particular, the presence of a detrimental low temperature Pt-Al reaction is identified. This is implicated in both the requirement for a higher Al:Ti ratio than is required for related AuPdAlTi contact schemes and through the degraded temperature dependent resistance behaviour of the annealed AuPtAlTi contacts
Heterocellular Crosstalk Between Myofibroblasts and Cardiomyocytes: Arrhythmogenic Effects of Transforming Growth Factor- 1
Effects of TGF- 1 on Electrophysiological Properties of Hybrid Cardiomyocyte-Myofibroblast Preparations
Effects of Cytokines on Electrophysiological Properties of Hybrid Cardiomyocyte-Myofibroblast Preparations
Arrhythmogenic Effects of Transforming Growth Factor- 1 on Hybrid Cardiomyocyte-Myofibroblast Preparations
The natural cardioprotective particle HDL modulates connexin43 gap junction channels
High-density lipoprotein (HDL) is known for its cardioprotective properties independent from its cholesterol transport activity. These properties are mediated by activation of kinases such as protein kinase C (PKC). Connexin43 (Cx43) is a gap junction protein present in ventricular cardiomyocytes. PKC-dependent phosphorylation modifies Cx43 gap junction channel properties and is involved in cardioprotection. We hypothesized that cardioprotective properties of HDL may be mediated in part by affecting Cx43 gap junction channels
- …