ABSTRACT
The aim of the present paper was to study the role of sodium calcium exchanger (NCX) in the generation of action potentials (APs) in cardiomyocytes during early developmental stage (EDS). The precisely dated embryonic hearts of C57 mice were dissected and enzymatically dissociated to single cells. The changes of APs were recorded by whole-cell patch-clamp technique before and after administration of NCX specific blockers KB-R7943 (5 μmol/L) and SEA0400 (1 μmol/L). The results showed that, both KB-R7943 and SEA0400 had potent negative chronotropic effects on APs of pacemaker-like cells, while such effects were only observed in some ventricular-like cardiomyocytes. The negative chronotropic effect of KB-R7943 on ventricular-like cardiomyocytes was accompanied by shortening of AP duration (APD), whereas such an effect of SEA0400 was paralleled by decrease in velocity of diastolic depolarization (Vdd). From embryonic day 9.5 (E9.5) to E10.5, the negative chronotropic effects of KB-R7943 and SEA0400 on ventricular-like APs of embryonic cardiomyocytes gradually disappeared. These results suggest that, in the short-term development of early embryo, the function of NCX may experience developmental changes as evidenced by different roles of NCX in autorhythmicity and APs generation, indicating that NCX function varies with different conditions of cardiomyocytes.
Subject(s)
Animals , Mice , Action Potentials , Calcium/metabolism , Myocytes, Cardiac/metabolism , Sodium/metabolism , Sodium-Calcium Exchanger , Thiourea/pharmacologyABSTRACT
Myocytes in the pulmonary veins (PV) play a pivotal role in the development of paroxysmal atrial fibrillation (AF). It is therefore important to understand physiological characteristics of these cells. Studies on these cells are, however, markedly impeded by the fact that single PV myocytes are very difficult to obtain due to lack of effective isolation methods. In this study, we described a novel PV myocyte isolation method. The key aspect of this method is to establish a combination of retrograde heart perfusion (via the aorta) and anterograde PV perfusion (via the pulmonary artery). With this simultaneous perfusion method, a better perfusion of the PV myocytes can be obtained. As results, the output and viability of single myocytes isolated by simultaneous heart and PV perfusion method were increased compared with those in conventional retrograde heart perfusion method.
Subject(s)
Animals , Rabbits , Atrial Fibrillation , Cell Separation , Heart , Muscle Cells , Perfusion , Pulmonary VeinsABSTRACT
Thymosin β4 (Tβ4) is a key factor in cardiac development, growth, disease, epicardial integrity, blood vessel formation and has cardio-protective properties. However, its role in murine embryonic stem cells (mESCs) proliferation and cardiovascular differentiation remains unclear. Thus we aimed to elucidate the influence of Tβ4 on mESCs. Target genes during mESCs proliferation and differentiation were detected by real-time PCR or Western blotting, and patch clamp was applied to characterize the mESCs-derived cardiomyocytes. It was found that Tβ4 decreased mESCs proliferation in a partial dose-dependent manner and the expression of cell cycle regulatory genes c-myc, c-fos and c-jun. However, mESCs self-renewal markers Oct4 and Nanog were elevated, indicating the maintenance of self-renewal ability in these mESCs. Phosphorylation of STAT3 and Akt was inhibited by Tβ4 while the expression of RAS and phosphorylation of ERK were enhanced. No significant difference was found in BMP2/BMP4 or their downstream protein smad. Wnt3 and Wnt11 were remarkably decreased by Tβ4 with upregulation of Tcf3 and constant β-catenin. Under mESCs differentiation, Tβ4 treatment did not change the expression of cardiovascular cell markers α-MHC, PECAM, and α-SMA. Neither the electrophysiological properties of mESCs-derived cardiomyocytes nor the hormonal regulation by Iso/Cch was affected by Tβ4. In conclusion, Tβ4 suppressed mESCs proliferation by affecting the activity of STAT3, Akt, ERK and Wnt pathways. However, Tβ4 did not influence the in vitro cardiovascular differentiation.
Subject(s)
Animals , Mice , Cell Cycle , Genetics , Cell Differentiation , Cell Movement , Cell Proliferation , Dose-Response Relationship, Drug , Extracellular Signal-Regulated MAP Kinases , Genetics , Metabolism , Gene Expression Regulation , JNK Mitogen-Activated Protein Kinases , Genetics , Metabolism , Mouse Embryonic Stem Cells , Cell Biology , Metabolism , Myocytes, Cardiac , Cell Biology , Metabolism , Nanog Homeobox Protein , Genetics , Metabolism , Octamer Transcription Factor-3 , Genetics , Metabolism , Patch-Clamp Techniques , Primary Cell Culture , Proto-Oncogene Proteins c-akt , Genetics , Metabolism , Proto-Oncogene Proteins c-fos , Genetics , Metabolism , Proto-Oncogene Proteins c-myc , Genetics , Metabolism , STAT3 Transcription Factor , Genetics , Metabolism , Signal Transduction , Thymosin , PharmacologyABSTRACT
<p><b>AIM</b>To investigate the influence of platelet-activating factor (PAF) receptor on long-term potentiation (LTP) attenuated by aluminium.</p><p><b>METHODS</b>The method of extracellular recording was used to investigate the effect of PAF receptors on PP-CA3 LTP by microinjection of PAF receptor antagonist Ginkgolide B or agonist mc-PAF into CA3 area.</p><p><b>RESULTS</b>(1) Amplitude of population spikes (PS) of evoked potential was not affected but LTP induction was blocked by 0.2 micromol/L ginkgolide B in CA3 area. (2) LTP induction was not influenced by 0.25 mol/L aluminium chloride, however, it could be blocked when aluminium was applicated with ginkgolide B. (3) LTP induction was influenced slightly by 40 micromol/L mc-PAF but it has no difference in statistic. LTP induction could be blocked completely by 0.5 mol/L aluminium, but when aluminium was coapplicated with mc-PAF, this effect could be relieved.</p><p><b>CONCLUSION</b>These results indicate that PAF receptors are involved in induction of LTP in CA3 area by stimulating perforant path. The inhibitory effect of aluminium on LTP is partly related to PAF receptors.</p>
Subject(s)
Animals , Rats , Aluminum Compounds , Toxicity , CA3 Region, Hippocampal , Metabolism , Electric Stimulation , Evoked Potentials , Ginkgolides , Pharmacology , Lactones , Pharmacology , Long-Term Potentiation , Perforant Pathway , Platelet Membrane Glycoproteins , Metabolism , Rats, Sprague-Dawley , Receptors, G-Protein-Coupled , MetabolismABSTRACT
Objective To study the effects of granisetron hydrochloride on vasovagal syncope(VVS) in rabbits.Methods Twenty-four healthy New Zealand rabbits were divided stochastically into control group and intervention group,12 in each group. The control group was injected intravenously with normal saline. The intervention group was injected intravenously with granisetron hydrchloiride.Rabbit VVS models were established,each was taken at 4 points in time in the bloodletting process:T1,T2,T3,T4,to compare the bloodletting time,the concentration of 5-hydroxytryptamine(5-HT) in T2,T3,T4 and the total blood volume between the groups,and monitor the heart rate, blood pressure during the entire process.Results 1.The time of intervention group in T2,T3,T4 was longer than the time of control group obviously(P