Altered vascular smooth muscle function in the ApoE knockout mouse during the progression of atherosclerosis

Objectives: Relaxation of vascular smooth muscle (VSM) requires re-uptake of cytosolic Ca2+ into the sarcoplasmic reticulum (SR) via the Sarco/Endoplasmic Reticulum Ca2+ ATPase (SERCA), or extrusion via the Plasma Membrane Ca2+ ATPase (PMCA) or sodium Ca2+ exchanger (NCX). Peroxynitrite, a reactive species formed in vascular inflammatory diseases, upregulates SERCA activity to induce relaxation but, chronically, can contribute to atherogenesis and altered vascular function by escalating endoplasmic reticulum stress. Our objectives were to determine if peroxynitrite-induced relaxation and Ca2+ handling processes within vascular smooth muscle cells were altered as atherosclerosis develops.<p></p> Methods: Aortae from control and ApoE−/− mice were studied histologically, functionally and for protein expression levels of SERCA and PMCA. Ca2+ responses were assessed in dissociated aortic smooth muscle cells in the presence and absence of extracellular Ca2+.<p></p> Results: Relaxation to peroxynitrite was concentration-dependent and endothelium-independent. The abilities of the SERCA blocker thapsigargin and the PMCA inhibitor carboxyeosin to block this relaxation were altered during fat feeding and plaque progression. SERCA levels were progressively reduced, while PMCA expression was upregulated. In ApoE−/− VSM cells, increases in cytosolic Ca2+ [Ca2+]c in response to SERCA blockade were reduced, while SERCA-independent Ca2+ clearance was faster compared to control.<p></p> Conclusion: As atherosclerosis develops in the ApoE−/− mouse, expression and function of Ca2+ handling proteins are altered. Up-regulation of Ca2+ removal via PMCA may offer a potential compensatory mechanism to help normalise the dysfunctional relaxation observed during disease progression

FK506 binding proteins : cellular regulators of intracellular Ca2+ channels

In many cell types the intracellular Ca2+ store performs a central role in the regulation of the cytosolic Ca2+ concentration ([Ca2+]c) , the elevation of which triggers diverse and fundamental activities from reproduction to apoptosis, as well as being the major triggr for contraction.Two distinct classses of Ca2+ release channels, which mobilize Ca2+ from the store, exist: the inositol 1,4,5-trisphosphate (IP3) receptor and the ryanodine receptor. Considerable attention has been directed towards the importance of modulatory proteins that interact with these channels including, FK506 binding proteins (FKBPs), FKBP12 and its isoform, FKBP12.6. Although FKBP12 was first identified as the principal intracellular target for the immunosuppressive drugs, FK506 and rapamycin, new insights into the role of FKBPs have since emerged. These regulatory proteins are reportedly important modulators of intracellular Ca2+ release. FKBPs may regulate ryanodine and IP3 receptors either directly, by binding to the cytoplasmic aspect of the channel, or indirectly via modulation of two targets, the phosphatase, calcineurin or the kinase, mammalian target of rapamycin (mTOR). Dissociation of FKBP12 or FKBP12.6 from either Ca2+ release channel may increase, decrease or have no effect on ryanodine receptor- or IP3 receptor-mediated Ca2+ release. These important controversies may be attributed to FKBPs' ability to regulate the receptor indirectly via the kinase and phosphatase pathways modulated by the accessory proteins. This brief review discusses the regulation of intracellular ryanodine and IP3 receptor Ca2+ release channels by accessory FKBPs, with important implications for the role of FKBPs in the pathophysiology of a number of diseases

Role of the sarcoplasmic reticulum in nitric oxide induced modulation of cytoplasmic calcium in rabbit aortic smooth muscle cells

SIGLEAvailable from British Library Document Supply Centre- DSC:DXN055882 / BLDSC - British Library Document Supply CentreGBUnited Kingdo

Developing learning landscapes: academic libraries driving organisational change

Purpose – The purpose of this paper is to discuss the reasons and drivers for academic libraries affecting university strategy with regards to shaping and developing learning spaces in response to changing pedagogic behaviours. Design/methodology/approach – A review of available literature within the context of academic libraries and their position to influence and lead institutional strategic change. This theory and practice is addressed and evidenced by four case studies of university libraries in the UK. Findings – Many UK academic libraries find themselves able to lead on and influence their institution's strategic direction with regards to teaching, learning and research. This is particularly the case in the design and development of learning spaces within the university. Academic libraries are in a unique position within a university with a view to observing student behaviours, being responsive to ever changing demands from academics and students, spotting trends and benchmarking against comparative institutions. These practices make it possible for academic libraries to advise, guide and lead on teaching and learning strategy and lead on learning spaces developments within their institutions. Practical implications – Academic libraries can use existing quality assurance, responsiveness and benchmarking frameworks to influence university strategy and decision making. Originality/value – This paper focuses on the concept of academic libraries influencing change, rather than responding to change, within their university. The case studies provide examples of where this has been the case, and suggest ways and frameworks which can be adopted by other academic libraries

Potassium channels underlying the resting potential of pulmonary artery smooth muscle cells

1. The molecular identity of the K channels giving rise to the negative membrane potential of pulmonary artery smooth muscle cells has yet to be determined. 2. To date, most studies have focused on voltage-gated, delayed rectifier channels and their roles in mediating hypoxia-induced membrane depolarization. There is, however, strong evidence that an outwardly rectifying K+ conductance distinct from the classical delayed rectifier is involved. 3. Growing evidence that TASK-like channels can sense hypoxia and are present in pulmonary artery smooth muscle cells suggests that they may be responsible for the resting K+ conductance and resting potential. 4. The present review considers the evidence that particular K channels maintain the resting membrane potential of pulmonary artery smooth muscle cells and mediate the depolarizing response to hypoxia

Sarcolemma agonist-induced interactions between InsP3 and ryanodine receptors in Ca2+ oscillations and waves in smooth muscle

Smooth muscle cells respond to InsP(3)-generating (sarcolemma-acting) neurotransmitters and hormones by releasing Ca(2+) from the internal store. However, the release of Ca(2+) does not occur uniformly throughout the cytoplasm but often into a localized area before being transmitted to other regions of the cell in the form of Ca(2+) waves and oscillations to actively spread information within and between cells. Yet, despite their significance, our understanding of the generation of oscillations to waves is incomplete. A major aspect of controversy centres on whether or not Ca(2+) released from the InsP(3) receptor activates RyRs (ryanodine receptors) to generate further release by Ca(2+)-induced Ca(2+) release and propagate waves or whether the entire process arises from InsP(3) receptor activity alone. Under normal physiological conditions the [Ca(2+)] required to activate RyR (approx. 15 microM) exceeds the bulk average [Ca(2+)](c) (cytoplasmic Ca(2+) concentration) generated by InsP(3) receptor activity (300 nM), features that favour intermittent activity of the receptor as occurs in waves and oscillations. Experimental evidence for the involvement of RyR relies mainly on pharmacological approaches in the intact cell where poor drug specificity could have led to ambiguous results. In this brief review the possible interactions between InsP(3) receptors and RyR in the generation of oscillations and waves will be discussed. Evidence is presented that RyRs are not required for InsP(3)-mediated Ca(2+) transients. Notwithstanding, ryanodine can inhibit InsP(3)-mediated Ca(2+) responses after RyR activity has been induced by caffeine or by steady depolarization which evokes spontaneous transient outward currents (a sarcolemmal manifestation of RyR activity). Ryanodine inhibits InsP(3)-mediated Ca(2+) transients by depleting the store of Ca(2+) rather than by RyR involvement in the InsP(3)-mediated Ca(2+) increase

The sarcoplasmic reticulum, Ca2+ trapping, and wave mechanisms in smooth muscle

The sarcoplasmic reticulum (SR) and apposed regions of the sarcolemma passively trap Ca2+ entering the cell to limit the rise in cytoplasmic Ca2+ concentration without SR pump involvement. When "leaky," the SR facilitates Ca2+ entry to the cytoplasm. SR Ca2+ release via inositol 1,4,5-trisphosphate receptors (IP(3)Rs) propagates as calcium waves; IP(3)Rs alone account for wave propagation

Ca2+ microdomains in smooth muscle

In smooth muscle, Ca2+ controls diverse activities including cell division, contraction and cell death. Of particular significance in enabling Ca2+ to perform these multiple functions is the cell's ability to localize Ca2+ signals to certain regions by creating high local concentrations of Ca2+ (microdomains), which differ from the cytoplasmic average. Microdomains arise from Ca2+ influx across the plasma membrane or release from the sarcoplasmic reticulum (SR) Ca2+ store. A single Ca2+ channel can create a microdomain of several micromolar near (not, vert, similar200 nm) the channel. This concentration declines quickly with peak rates of several thousand micromolar per second when influx ends. The high [Ca2+] and the rapid rates of decline target Ca2+ signals to effectors in the microdomain with rapid kinetics and enable the selective activation of cellular processes. Several elements within the cell combine to enable microdomains to develop. These include the brief open time of ion channels, localization of Ca2+ by buffering, the clustering of ion channels to certain regions of the cell and the presence of membrane barriers, which restrict the free diffusion of Ca2+. In this review, the generation of microdomains arising from Ca2+ influx across the plasma membrane and the release of the ion from the SR Ca2+ store will be discussed and the contribution of mitochondria and the Golgi apparatus as well as endogenous modulators (e.g. cADPR and channel binding proteins) will be considered

Media 1: Light-induced Ca²⁺ transients observed in widefield epi-fluorescence microscopy of excitable cells

Originally published in Biomedical Optics Express on 01 June 2012 (boe-3-6-1266