A novel secretagogue increases cardiac contractility by enhancement of L-type Ca2+ current
N'1-(3,3,6,8-tetramethyl-1-oxo-1,2,3,4-tetrahydronaphthalen-2-yliden)-2-cyanoethanohydrazide (TTYC) increases secretion of glucagon-like peptide-1 and intracellular Ca(2+) concentration in GLUTag cells. The purpose of the present study was to examine if TTYC exerts positive inotropic effects on isolated rabbit ventricular myocytes and in vivo heart in anesthetized rats, and if so to further define the potential mechanism of action. Contractility was assessed in vitro using changes in fractional shortening (FS) of myocyte sarcomere length and in vivo using changes in the velocity of left ventricular pressure. Changes in L-type Ca(2+) current of ventricular myocytes were evaluated using whole-cell voltage-clamp techniques. TTYC increased FS of myocyte sarcomere length in a concentration-dependent manner. The positive inotropic effect was not abrogated by beta-adrenergic blockade (propranolol) or protein kinase A inhibition. TTYC enhanced peak L-type Ca(2+) current in a voltage-dependent manner (current amplitudes increased by 4.0-fold at -10 mV and 1.5-fold at +10 mV). Voltage-dependence of steady-state activation of L-type Ca(2+) current was shifted by 15 mV in the negative direction. Inactivation time course of the L-type Ca(2+) currents at voltages of -10 to 20 mV was significantly slowed by 0.3 microM TTYC. In vivo studies demonstrated that TTYC increased cardiac contractility in a dose-dependent manner. In conclusion, TTYC is a novel L-type Ca(2+) current activator with positive cardiac inotropic effects. Negative shifting of the voltage-dependence of L-type Ca(2+) current activation and reduced inactivation are two mechanisms responsible for the enhanced L-type Ca(2+) current that contribute to the positive inotropic effects.
[Show abstract] [Hide abstract] ABSTRACT: Chronic psychosocial stress triggers cardiovascular diseases although underlying mechanisms are still elusive. This study examined the effect of social stress on cardiomyocyte contractile function and pathological changes in myocardium using the visible burrow system (VBS) model. Chronic social stress was induced using a mixed-sex VBS housing in adult Sprague-Dawley (SD) rats. Contractile and intracellular Ca(2+) properties were evaluated in isolated cardiomyocytes including peak shortening (PS), time-to-PS (TPS), time-to-90% relengthening (TR(90)), maximal velocity of shortening/relengthening (± dL/dt), Fura-2 fluorescence intensity, and intracellular Ca(2+) decay. Myocardial histology was evaluated using Masson trichrome staining. Social stress led to depressed PS, ± dL/dt, shortened TPS and prolonged TR(90) compared with the unstressed controls. Baseline and electrically-stimulated rise in Ca(2+) were reduced whereas intracellular Ca(2+) decay was delayed in stressed rats. Histological analyses exhibited overt interstitial fibrosis and cardiomyocyte hypertrophy in stressed rats. The GSH/GSSG ratio (indicative of oxidative stress status) was reduced whereas oxidative protein carbonyl formation was elevated in stressed rats. Western blot analysis showed unchanged expression of superoxide dismutase 1 (SOD1), β(1)-adrenoceptor (β(1)-AR) levels, reduced sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA2a) levels, and elevated phosphorylation of the stress signaling protein kinase JNK but not ERK in myocardium from stressed rats. Short-term in vitro treatment of cardiomyocytes with the stress inducer phenylephrine mimicked cell damage and intracellular Ca(2+) mishandling, the effects of which were mitigated by antioxidant, JNK inhibition, carvedilol and SERCA2a adenovirus. These findings indicate that chronic social stress is detrimental to cardiac structure and function possibly via mechanisms associated with oxidative injury and intracellular Ca(2+) mishandling.0Comments 2Citations
- "To examine the effect of antioxidant and adrenergic inhibition in stress-induced cardiac mechanical derangement, isolated cardiomyocytes from control unstressed rats were exposed to the stress inducer phenylephrine (20 µM)  to mimic a stress environment in the absence or presence of the antioxidant NAC (500 µM)  the mixed α-/β-adrenergic antagonist carvedilol (100 nM) , or the β-adrenergic antagonist propanolol (1 µM) . Phenylephrine incubation significantly interrupted intracellular Ca 2+ homeostasis as evidenced by decreased ΔFFI and peak FFI as well as prolonged intracellular Ca 2+ decay (single exponential) associated with unchanged baseline FFI levels, reminiscent of the intracellular Ca 2+ defect observed in socially stressed rat cardiomyocytes. "
[Show abstract] [Hide abstract] ABSTRACT: Cardiac toxicity is a major concern in drug development and it is imperative that clinical candidates are thoroughly tested for adverse effects earlier in the drug discovery process. In this report, we investigate the utility of an impedance-based microelectronic detection system in conjunction with mouse embryonic stem cell-derived cardiomyocytes for assessment of compound risk in the drug discovery process. Beating of cardiomyocytes was measured by a recently developed microelectronic-based system using impedance readouts. We used mouse stem cell-derived cardiomyocytes to obtain dose-response profiles for over 60 compounds, including ion channel modulators, chronotropic/ionotropic agents, hERG trafficking inhibitors and drugs known to induce Torsades de Pointes arrhythmias. This system sensitively and quantitatively detected effects of modulators of cardiac function, including some compounds missed by electrophysiology. Pro-arrhythmic compounds produced characteristic profiles reflecting arrhythmia, which can be used for identification of other pro-arrhythmic compounds. The time series data can be used to identify compounds that induce arrhythmia by complex mechanisms such as inhibition of hERG channels trafficking. Furthermore, the time resolution allows for assessment of compounds that simultaneously affect both beating and viability of cardiomyocytes. Microelectronic monitoring of stem cell-derived cardiomyocyte beating provides a high throughput, quantitative and predictive assay system that can be used for assessment of cardiac liability earlier in the drug discovery process. The convergence of stem cell technology with microelectronic monitoring should facilitate cardiac safety assessment.0Comments 48Citations
- "The respective values obtained here are within the range reported for these compounds using electrophysiological methods and primary cardiomyocytes or human ES cellderived cardiomyocytes (Supporting InformationTable S2 and Denning and Anderson, 2008). However, the IC 50 values obtained by patch clamp in cells transfected with the hERG channel appear to be lower (Su et al., 2010; Gintant, 2011). In addition to cell type differences and levels of ERG channel expression (or hERG channel in the case of heterologous expression in CHO cells), the differences could be explained by the workflow of the two approaches. "
- [Show abstract] [Hide abstract] ABSTRACT: In this paper, we present an investigation of GaAs MMIC semiconductor limiters which can be fabricated on a standard MMIC process. This investigation, which included high power vector measurements of S-parameters over lime, led to the design of a highly compact limiter integrated directly onto an FM-CW radar MMIC. As we demonstrate, the limiter significantly improved the lifetime of the FM-CW radar MMIC when exposed to both single and multiple short, high power pulses of microwave energy0Comments 6Citations