Three-Week-Old Rabbit Ventricular Cardiomyocytes as a Novel System to Study Cardiac Excitation and EC Coupling.
Three-Week-Old Rabbit Ventricular Cardiomyocytes as a Novel System to Study Cardiac Excitation and EC Coupling.
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DOI：
10.3389/fphys.2021.672360
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发表时间：
2021
影响因子：
4
通讯作者：
Koren G
中科院分区：
文献类型：
--
作者：
Kabakov AY;Sengun E;Lu Y;Roder K;Bronk P;Baggett B;Turan NN;Moshal KS;Koren G
心律失常是心血管疾病发病率和死亡率的重要原因。兔心脏是研究心肌细胞兴奋和致心律失常的公认模型系统。因此，成年兔心室肌细胞的原代培养可作为研究人类心脏兴奋的分子机制的优选模型。然而，使用成年兔心肌细胞通常被认为是过于昂贵的。因此，我们开发并表征了一种新的低成本兔心肌细胞模型，即3周龄心室心肌细胞（3 wRbCMs）。通过标准酶技术从3周龄新西兰白色兔（两种性别）的整个心室中分离心室肌细胞。使用麦胚凝集素，我们发现了一个明确的T-小管结构急性分离的3 wRbCMs。用腺病毒感染细胞（感染复数为10）以表达绿色荧光蛋白（GFP）并培养48小时。细胞的动作电位时程（APD 90 = 253 ± 24 ms）和钙瞬变与成年兔心肌细胞相似。新鲜分离和48 h培养的细胞表达关键离子通道蛋白：钙电压门控通道亚基α 1 C（Cavα1c）、钠电压门控通道α亚基5（Nav1.5）、钾电压门控通道亚家族D成员3（Kv4.3）和亚家族A成员4（Kv1.4）以及亚家族H成员2（RERG. Kv11.1）、KvLQT 1（K7.1）蛋白和内向整流钾通道（Kir2.1）。细胞表现出适当的电生理表型，包括快钠电流（INa），瞬时外向钾电流（Ito），L型钙通道峰值电流（伊卡，L），延迟整流钾电流的快速和缓慢成分（IKr和IKs），和内向整流（IK 1）。虽然通道蛋白和一些电流的表达在48小时的培养过程中下降，我们得出结论，3 wRbCMs是一种新的，低成本的替代成年兔心肌细胞系统，它允许调查的分子机制的心脏兴奋的形态，生化，遗传，生理和生物物理水平。
Cardiac arrhythmias significantly contribute to cardiovascular morbidity and mortality. The rabbit heart serves as an accepted model system for studying cardiac cell excitation and arrhythmogenicity. Accordingly, primary cultures of adult rabbit ventricular cardiomyocytes serve as a preferable model to study molecular mechanisms of human cardiac excitation. However, the use of adult rabbit cardiomyocytes is often regarded as excessively costly. Therefore, we developed and characterized a novel low-cost rabbit cardiomyocyte model, namely, 3-week-old ventricular cardiomyocytes (3wRbCMs). Ventricular myocytes were isolated from whole ventricles of 3-week-old New Zealand White rabbits of both sexes by standard enzymatic techniques. Using wheat germ agglutinin, we found a clear T-tubule structure in acutely isolated 3wRbCMs. Cells were adenovirally infected (multiplicity of infection of 10) to express Green Fluorescent Protein (GFP) and cultured for 48 h. The cells showed action potential duration (APD90 = 253 ± 24 ms) and calcium transients similar to adult rabbit cardiomyocytes. Freshly isolated and 48-h-old-cultured cells expressed critical ion channel proteins: calcium voltage-gated channel subunit alpha1 C (Cavα1c), sodium voltage-gated channel alpha subunit 5 (Nav1.5), potassium voltage-gated channel subfamily D member 3 (Kv4.3), and subfamily A member 4 (Kv1.4), and also subfamily H member 2 (RERG. Kv11.1), KvLQT1 (K7.1) protein and inward-rectifier potassium channel (Kir2.1). The cells displayed an appropriate electrophysiological phenotype, including fast sodium current (INa), transient outward potassium current (Ito), L-type calcium channel peak current (ICa,L), rapid and slow components of the delayed rectifier potassium current (IKr and IKs), and inward rectifier (IK1). Although expression of the channel proteins and some currents decreased during the 48 h of culturing, we conclude that 3wRbCMs are a new, low-cost alternative to the adult-rabbit-cardiomyocytes system, which allows the investigation of molecular mechanisms of cardiac excitation on morphological, biochemical, genetic, physiological, and biophysical levels.
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影响因子：
5.8
作者：
Bartos DC;Grandi E;Ripplinger CM
通讯作者：
Ripplinger CM
DOI：
10.1016/j.bbamcr.2015.11.005
发表时间：
2016-07
期刊：
Biochimica et biophysica acta
影响因子：
--
作者：
Bedada FB;Wheelwright M;Metzger JM
通讯作者：
Metzger JM
影响因子：
20.1
作者：
Ackers-Johnson M;Li PY;Holmes AP;O'Brien SM;Pavlovic D;Foo RS
通讯作者：
Foo RS
影响因子：
5
作者：
Biet, Michael;Barajas-Martinez, Hector;Dumaine, Robert
通讯作者：
Dumaine, Robert
影响因子：
20.1
作者：
Dixon, JE;Shi, WM;McKinnon, D
通讯作者：
McKinnon, D