Rune Sundset

Ischemia Induces Closure of Gap Junctional Channels and Opening of Hemichannels in Heart- derived Cells and Tissue

Aim: Gap junction intercellular communication (GJIC) and hemichannel permeability may have important roles during an ischemic insult. Our aim was to evaluate the effect of ischemia on gap junction channels and hemichannels. Methods: We used neonatal rat heart myofibroblasts and simulated ischemia with a HEPES buffer with high potassium, low pH, absence of glucose, and oxygen tension was reduced by dithionite. Microinjection, western blot, immunofluorescence, cell viability and dye uptake were used to evaluate the effects induced by dithionite. Isolated perfused rat hearts were used to analyse infarct size. Results: Short period with simulated ischemia reduced the ability to transfer a dye between neighbouring cells, which indicated reduced GJIC. Prolonged exposure to simulated ischemia caused opening of hemichannels, and cell death was apparent while gap junction channels remained closed. Connexin 43 became partially dephosphorylated and the total amount decreased during simulated ischemia. We were not able to detect the alternative hemichannel-forming protein, Pannexin 1, in these cells. The potential importance of Connexin 43 or Pannexin 1 hemichannels in ischemia-induced infarct in the intact heart was studied by perfusion of the heart in the presence of peptides that block one or the other type of hemichannels. The connexin-derived peptide, Gap26, significantly reduced the infract/risk zone ratio (control 48.7±4.2% and Gap26 19.4±4.1%, p<0.001), while the pannexin-derived peptide, 10Panx1, did not change infarct/risk ratio. Conclusion: Connexin 43 is most likely responsible for both closure of gap junction channels and opening of hemichannels during simulated ischemia in neonatal rat heart myofibroblasts. Opening of connexin 43 hemichannels during ischemia-reperfusion seems to be an important mechanism for ischemia-reperfusion injury in the heart. By preventing the opening of these channels during early ischemia-reperfusion the infarct size becomes significantly reduced.

Heptanol triggers cardioprotection via mitochondrial mechanisms and mitochondrial potassium channel opening in rat hearts

Aim:  To investigate mechanisms behind heptanol (Hp)‐induced infarct size reduction and in particular if protection by pre‐treatment with Hp is triggered through mitochondrial mechanisms.

Repeated simulated ischemia and protection against gap junctional uncoupling.

Ischemic preconditioning increases the hearts tolerance to a subsequent longer ischemic period. The aim of this study was to investigate the effect of early and delayed preconditioning on gap junction communication, connexin abundance, and phosphorylation in cultured neonatal rat cardiac myocytes. Prolonged ischemia followed 5 minutes after preconditioning in the early protocol, whereas 20 hours separated preconditioning and prolonged ischemia in the delayed preconditioning protocol. Gap junctional intercellular communication (GJIC) was assessed by Lucifer yellow dye transfer. An initial reduction in communication in response to sublethal ischemia was observed. This may be one mechanism whereby neighboring cells are protected from damaging substances produced during the first phase of subsequent regional ischemia in early preconditioning protocols. With respect to delayed preconditioning, the transient decrease in GJIC disappeared prior to prolonged ischemia, indicating that other mechanisms are responsible for delayed protection. Both early and delayed preconditioning preserved intercellular coupling after prolonged ischemia and this correlated with presence of less connexin43 dephosphorylation assessed by immunoblot.

Mechanism of hypoxic preconditioning in guinea pig papillary muscles

UNLABELLED Brief ischemia or hypoxia has been found to protect the heart against subsequent long-lasting ischemia and to improve contractile dysfunction as well to reduce cell necrosis and the incidence of lethal arrhythmias. This phenomenon, termed preconditioning (PC) has been demonstrated in different species. However, little is known about PC in guinea pigs. Moreover, electrophysiological changes underlying protection have not been studied so far in conjuntion with force recovery in a setting of PC. The aim of the study was to study PC in a guinea pig papillary muscle, using recovery of contractility after long hypoxic challenge as the main end-point of protection, and to investigate concomitant electrophysiological alterations. In guinea pig papillary muscle preparations contracting isometrically (paced at 2 Hz), transmembrane action potentials (AP) and developed force (DF) were recorded by conventional microelectrode technique and a force transducer. In addition, effective refractory periods (ERP) were determined. Hypoxia was induced by superfusion with 100% N2 (pO2 < 5 kPa) and pacing at 3,3 Hz. In the control group, long hypoxia lasted for 45 min and was followed by 30 min reoxygenation. In the PC group, muscles were subjected to 5 min hypoxia followed by 10 min recovery prior to sustained hypoxia/reoxygenation. RESULTS Long hypoxia induced a similar depression of DF in both, PC and control groups. However, a loss of contractile activity occured earlier in the PC group. AP duration and ERP decreased faster and were significantly shorter after PC. Upon reoxygenation, preconditioned muscles showed significantly better recovery of function (DF 86% of prehypoxic value vs. 36% in controls; p < 0,05). AP and ERP were completely restored in both, PC and control groups. Guinea pig papillary muscle can be preconditioned with a brief hypoxic challenge against contractile dysfunction upon long-lasting hypoxia/reoxygenation. Shortening of AP and loss of contractility occured more quickly during hypoxia and may participate in the protective effect of preconditioning. Possible mechanisms might involve facilitated opening of K(ATP)-dependent channels.

Ischemic preconditioning protects against gap junctional uncoupling in cardiac myofibroblasts

Ischemic preconditioning increases the hearts tolerance to a subsequent longer ischemic period. The purpose of this study was to investigate the role of gap junction communication in simulated preconditioning in cultured neonatal rat cardiac myofibroblasts. Gap junctional intercellular communication was assessed by Lucifer yellow dye transfer. Preconditioning preserved intercellular coupling after prolonged ischemia. An initial reduction in coupling in response to the preconditioning stimulus was also observed. This may protect neighboring cells from damaging substances produced during subsequent regional ischemia in vivo, and may preserve gap junctional communication required for enhanced functional recovery during subsequent reperfusion.

Mechanism of hypoxic preconditionin in guinea pig papillary muscles

Brief ischemia or hypoxia has been found to protect the heart against susbsequent long-lasting ischemia and to improve contractile dysfunction as well to reduce cell necrosis and the incidence of lethal arrhythmias. This phenomenon, termed preconditioning (PC) has been demonstrated in different species. However, little is known about PC in guinea pigs. Moreover, electrophysiological changes underlying protection have not been studied so far in conjuntion with force recovery in a setting of PC. The aim of the study was to study PC in a guinea pig papillary muscle, using recovery of contractility after long hypoxic challenge as the main end-point of protection, and to investigate concominant electrophysiological alterations. In guinea pig papillary muscle preparations contracting isometrically (paced at 2 Hz), transmembrane action potentials (AP) and developed force (DF) were recorded by conventional microelectrode technique and a force tranducer. In addition, effective refractory periods (ERP) were determined. Hypoxia was induced by superfusion with 100% N2 (pO2 < 5 kPa) and pacing at 3,3 Hz. In the control group, long hypoxia lasted for 45 min and was followed by 30 min reoxygenation. In the PC group, muscles were subjected to 5 min hypoxia followed by 10 min recovery prior to sustained hypoxia/reoxygenation. Results: Long hypoxia induced a similar depression of DF in both, PC and control groups. However, a loss of contractile activity occured earlier in the PC group. AP duration and ERP decreased faster and were significantly shorter after PC. Upon reoxygenation, preconditioned muscles showed significantly better recovery of function (DF 86% of prehypoxic value vs. 36% in controls; p < 0,05). AP and ERP were completely restored in both, PC and control groups. Guinea pig papillary muscle can be preconditioned with a brief hypoxic challenge against contractile dysfunction upon long-lasting hypoxia/reoxygenation. Shortening of AP and loss of contractility occured more quickly during hypoxia and may participate in the protective effect of preconditioning. Possible mechanisms might involve facilitated opening of KATP-dependent channels.

Nuclear medicine examination in heart failure patients

BACKGROUND Recently, cardiac resynchronization therapy, by using biventricular pacemakers, has become implemented in the treatment of patients with severe heart failure. However, using the classical inclusion criteria, 30 % of patients treated with resynchronization do not improve symptoms or activity level. Phase analysis of radionuclide ventriculography gives information about pattern of ventricular contraction and may detect dyssynchrony. The method may therefore be used to select patients with dyssynctrony to resynchronization therapy. In this study we have investigated the amount of dyssynchrony, by using phase analysis of radionuclide ventriculography, in men and women as a function of left ventricular ejection fraction. MATERIAL AND METHODS The study is based on 1 266 radionuclide ventriculographies performed at Section of Nuclear Medicine, University Hospital North-Norway, during 1998 - 2006. The relationship between left ventricular ejection fraction and number of patients with ventricular dyssynchrony was investigated. 90 patients with no known heart problems were considered as reference values for synchrony data. RESULTS The phase analysis showed that 35 % of the women and 34 % of the men with left ventricular ejection fraction below 35 % had both inter- and intra-ventricular dyssynchrony. INTERPRETATION Todays criteria for including patients to resynchonizing therapy are not good enough. Phase analysis of radionuclide ventriculography of patients with left ventricular ejection fraction below 35 % shows that one third of the patients have both inter- and intra- ventricular dyssynchrony. These patients might be responders to resynchronization therapy.

Connexin, connection, conductance: Towards understanding induction of arrhythmias?

Myocardial ischemia can result in three different forms of injury to the heart: arrhythmia, contractile dysfunction, and myocardial infarction. Arrhythmia and contractile dysfunction are reversible conditions that appear early (within few minutes) when ischemia occurs in a beating heart. Ventricular tachyarrhythmias are common and potentially life-threatening complications of myocardial ischemia and infarction. Although ventricular tachycardia at times can start by a nonreentrant mechanism (e.g., from delayed or early afterdepolarization), most runs of ventricular fibrillation during early myocardial ischemia are initiated by intramural reentry due to slow conduction block within the ischemic zone. 1 At the cellular level, the proposed events responsible for ischemic arrhythmias are calcium overload, partial depolarization of the cell membrane, and cellular uncoupling. Although advances in antiarrhythmic agents and implantation of direct-current defibrillators have resulted in improved prevention of death due to arrhythmia in myocardial ischemia, morbidity and mortality due to arrhythmias are still high. In the heart, an electrical impulse is propagated in the