University of Bern , Department of Physiology, Bühlplatz 5, CH-3012 Bern, Switzerland.
Neurophotonics. 2015 Apr;2(2):021011. doi: 10.1117/1.NPh.2.2.021011. Epub 2015 Feb 13.
We explore the feasibility of obtaining a spatially resolved picture of [Formula: see text] inward currents ([Formula: see text]) in multicellular cardiac tissue by differentiating optically recorded [Formula: see text] transients that accompany propagating action potentials. Patterned growth strands of neonatal rat ventricular cardiomyocytes were stained with the [Formula: see text] indicators Fluo-4 or Fluo-4FF. Preparations were stimulated at 1 Hz, and [Formula: see text] transients were recorded with high spatiotemporal resolution ([Formula: see text], 2 kHz analog bandwidth) with a photodiode array. Signals were differentiated after appropriate digital filtering. Differentiation of [Formula: see text] transients resulted in optically recorded calcium currents (ORCCs) that carried the temporal and pharmacological signatures of L-type [Formula: see text] inward currents: the time to peak amounted to [Formula: see text] (Fluo-4FF) and [Formula: see text] (Fluo-4), full-width at half-maximum was [Formula: see text], and ORCCs were completely suppressed by [Formula: see text][Formula: see text]. Also, and as reported before from patch-clamp studies, caffeine reversibly depressed the amplitude of ORCCs. The results demonstrate that the differentiation of [Formula: see text] transients can be used to obtain a spatially resolved picture of the initial phase of [Formula: see text] in cardiac tissue and to assess relative changes of activation/fast inactivation of [Formula: see text] following pharmacological interventions.
我们探讨了通过区分伴随传播动作电位的光记录的[Formula: see text]瞬变来获得细胞内电流([Formula: see text])在多细胞心脏组织中空间分辨图像的可行性。用[Formula: see text]指示剂 Fluo-4 或 Fluo-4FF 对新生大鼠心室心肌细胞的图案化生长链进行染色。以 1 Hz 的频率刺激制剂,并使用光电二极管阵列以高时空分辨率([Formula: see text],2 kHz 模拟带宽)记录[Formula: see text]瞬变。对适当的数字滤波后的信号进行区分。[Formula: see text]瞬变的区分导致光记录的钙电流(ORCC),其具有 L 型[Formula: see text]内向电流的时间和药理学特征:峰值时间达到[Formula: see text](Fluo-4FF)和[Formula: see text](Fluo-4),半最大值全宽为[Formula: see text],并且 ORCC 被[Formula: see text][Formula: see text]完全抑制。此外,正如之前从膜片钳研究中报道的那样,咖啡因可逆地抑制了 ORCC 的幅度。结果表明,[Formula: see text]瞬变的区分可用于获得心脏组织中[Formula: see text]初始阶段的空间分辨图像,并评估药物干预后[Formula: see text]的激活/快速失活的相对变化。