ABSTRACT
Understanding developmental changes in contractility is critical to improving therapies for young cardiac patients. Isometric developed force was measured in human ventricular muscle strips from two age groups: newborns (<2 wk) and infants (3-14 mo) undergoing repair for congenital heart defects. Muscle strips were paced at several cycle lengths (CLs) to determine the force frequency response (FFR). Changes in Na/Ca exchanger (NCX), sarcoplasmic reticulum Ca-ATPase (SERCA), and phospholamban (PLB) were characterized. At CL 2000 ms, developed force was similar in the two groups. Decreasing CL increased developed force in the infant group to 131 +/- 8% (CL 1000 ms) and 157 +/- 18% (CL 500 ms) demonstrating a positive FFR. The FFR in the newborn group was flat. NCX mRNA and protein levels were significantly larger in the newborn than infant group whereas SERCA levels were unchanged. PLB mRNA levels and PLB/SERCA ratio increased with age. Immunostaining for NCX in isolated newborn cells showed peripheral staining. In infant cells, NCX was also found in T-tubules. SERCA staining was regular and striated in both groups. This study shows for the first time that the newborn human ventricle has a flat FFR, which increases with age and may be caused by developmental changes in calcium handling.
Subject(s)
Heart Defects, Congenital/physiopathology , Heart Ventricles/physiopathology , Muscle Strength , Myocardial Contraction , Ventricular Function , Age Factors , Calcium Signaling , Calcium-Binding Proteins/metabolism , Cardiac Pacing, Artificial , Cardiac Surgical Procedures , Female , Heart Defects, Congenital/metabolism , Heart Defects, Congenital/surgery , Heart Ventricles/growth & development , Heart Ventricles/metabolism , Humans , In Vitro Techniques , Infant , Infant, Newborn , Male , RNA, Messenger/metabolism , Sarcoplasmic Reticulum Calcium-Transporting ATPases/metabolism , Sodium-Calcium Exchanger/metabolismABSTRACT
Dopamine is used to treat heart failure, particularly after cardiac surgery in infants, but the mechanisms of action are unclear. We investigated differences in the effect of dopamine on L-type calcium current (I(Ca)) between newborn (NB, 1-4 days) and adult (AD, 3-4 mo) rabbit ventricular myocytes. Myocytes were enzymatically dissociated from NB and AD rabbit hearts. I(Ca) was recorded by using the whole cell patch-clamp technique. mRNA levels of cardiac dopamine receptor type 1 (D1), type 2 (D2), and beta-adrenergic receptors (beta-ARs) were measured by real-time RT-PCR. Dopamine (100 microM) increased I(Ca) more in NB (E(max) 87 +/- 10%) than in AD ventricular cells (E(max) 21 +/- 3%). Further investigation of this difference showed that mRNA levels of the D1 receptor were significantly higher in NB, and, with beta-AR blockade, dopamine increased I(Ca) more in NB than AD cells. Additionally, SKF-38393 (selective D1 receptor agonist) significantly increased I(Ca) by 55 +/- 4% in NB (P < 0.05, n = 4) and by 11 +/- 1% in AD (P < 0.05, n = 6). Dopamine in the presence of SCH-23390 (D1 receptor antagonist) increased I(Ca) in NB cells by 67 +/- 5% and by 22 +/- 2% in AD cells, suggesting a role for beta-AR stimulation. Selective blockade of beta(1)- or beta(2)-receptors (with block of D1 receptors) showed that the beta-AR action of dopamine in the NB was largely mediated via beta(2)-AR activation. Dopamine produces a larger increase in I(Ca) in NB cardiomyocytes compared with ADs. The mechanism of action is not only through beta(2)-ARs but also due to higher expression of cardiac D1 receptor in NB.
Subject(s)
Adrenergic beta-2 Receptor Agonists , Calcium Channels, L-Type/drug effects , Calcium/metabolism , Cardiotonic Agents/pharmacology , Dopamine/pharmacology , Myocytes, Cardiac/drug effects , Receptors, Dopamine D1/agonists , Signal Transduction/drug effects , Adrenergic beta-1 Receptor Agonists , Adrenergic beta-Antagonists/pharmacology , Age Factors , Animals , Animals, Newborn , Calcium Channels, L-Type/metabolism , Dopamine Agonists/pharmacology , Dopamine Antagonists/pharmacology , Membrane Potentials/drug effects , Myocytes, Cardiac/metabolism , Patch-Clamp Techniques , RNA, Messenger/metabolism , Rabbits , Receptors, Adrenergic, beta-2/genetics , Receptors, Adrenergic, beta-2/metabolism , Receptors, Dopamine D1/genetics , Receptors, Dopamine D1/metabolism , Receptors, Dopamine D2/agonists , Reverse Transcriptase Polymerase Chain Reaction , Up-RegulationABSTRACT
INTRODUCTION: Single-site ventricular pacing results in dyssynchronous ventricular activation and may contribute to ventricular dysfunction. We evaluated epicardial biventricular (BiV) pacing as a means of maintaining synchronous ventricular activation in an acute piglet model of AV block with normal ventricular anatomy and function. METHODS AND RESULTS: We used left ventricular (LV) impedance catheters and tissue Doppler imaging to assess the cardiodynamics of immature piglets (n = 6, 33-78 days, 9.35 +/- 0.85 kg). Following catheter ablation of the AV node, a pacemaker was programmed 20 beats per minute above the intrinsic atrial rate. The animals were paced at 5-minute intervals via the following AV sequential configurations: Right atrial appendage (RA)-RV apex (RVA), RA-LV apex (LVA), and RA-biventricular (RVA/LVA). RA-RVA was the experimental control. LV systolic mechanics, measured by the slope of the end-systolic pressure-volume relationship (E(es)), increased with BiV pacing (12.8 +/- 3.4 mmHg/mL, P < 0.02) or single-site LVA pacing (10.6 +/- 3.4 mmHg/mL, P < 0.05) compared with single-site RVA pacing (8.3 +/- 1.4 mmHg/mL). QRS duration lengthened compared with sinus rhythm (42 +/- 8 msec) with either RVA (56 +/- 9 msec, P < 0.02) or LVA (54 +/- 7 msec, P < 0.03), but not with BiV (48 +/- 7 msec, P = 0.08) pacing. Tissue Doppler imaging showed LV dyssynchrony with RVA (septal-to-lateral delay 46.0 +/- 51.7 msec), with return toward normal with LVA (-9.6 +/- 33.6 msec, P < 0.04) or BiV (-4.1 +/- 33.8 msec, P < 0.04) pacing. CONCLUSIONS: In this acute immature piglet model of AV block, LV performance improved with single-site pacing from the LVA and BiV pacing (RVA/LVA), as compared with single-site pacing from the RVA. These changes correlated with tissue Doppler indices of mechanical synchrony, though not necessarily with QRS duration.
Subject(s)
Cardiac Pacing, Artificial/methods , Disease Models, Animal , Heart Block/therapy , Ventricular Function, Left , Animals , Echocardiography , Elasticity , Electrocardiography , Heart Block/physiopathology , Swine , SystoleABSTRACT
Increased ventricular expression of genes encoding for various structural and contractile proteins has been reported in cardiac hypertrophy. Mechanisms leading to this altered gene expression are only partly understood. Recently, various transcription factors (TF), among them GATA-4, Nkx-2.5/Csx, MEF-2 and the HAND family (eHAND and dHAND), and their role in embryonic cardiac development have been described. These transcription factors are known to have binding sites to promotor regions of many genes known to be regulated in hypertrophy of adult ventricular myocardium. We investigated the temporal and spatial expression pattern of these transcription factors in a rat model of acute pressure-overload of the right ventricle, induced by banding (coarctation) of the pulmonary artery. Expression of GATA-4, Nkx-2.5/Csx, MEF-2 and dHAND protein was found to increase in the right ventricle after the banding procedure as determined by immunohistochemistry and western blotting. A marker of the onset of cardiac hypertrophy was expression of ANP protein. We conclude that TF known to regulate embryonic heart development are involved in the adaptational response of adult ventricular myocardium to pressure overload.
