Silencing MaxiK Activity in Corporal Smooth Muscle Cells Initiates Compensatory Mechanisms to Maintain Calcium Homeostasis
Silencing MaxiK Activity in Corporal Smooth Muscle Cells Initiates Compensatory Mechanisms to Maintain Calcium Homeostasis
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DOI：
10.1111/j.1743-6109.2010.02180.x
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发表时间：
2011-08-01
影响因子：
3.5
通讯作者：
Davies, Kelvin Paul
中科院分区：
文献类型：
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作者：
Calenda, Giulia;Suadicani, Sylvia Ottilie;Davies, Kelvin Paul
介绍。 MaxiK 钾通道受电压和细胞内钙离子的调节，在调节细胞内钙离子浓度 ([Ca(2+)](i)) 中起关键作用，而细胞内钙离子浓度是平滑肌张力的最终决定因素。严格控制海绵体平滑肌 (CCSM) 张力至关重要，调节不当可能导致勃起功能障碍。由于 MaxiK 和钙通道活性的紧密功能联系，本研究的目的是确定 MaxiK 的沉默和药理学抑制对 CCSM 细胞中钙稳态和细胞间钙信号传导的影响。方法。我们比较了 MaxiK 沉默 CCSM 细胞 48 小时内基础细胞内 [Ca(2+)](i) 和定义细胞间钙波 (ICW) 传播的参数与伊贝里奥毒素急性阻断通道的变化。为了分析基因表达中发生的变化，我们在 MaxiK 沉默 48 小时后进行了微阵列分析。主要结果测量。测量 Fura-2 荧光强度的变化以评估基础 [Ca(2+)](i) 水平和 ICW 参数。对整体基因表达进行微阵列分析。结果。 MaxiK 沉默基础 [Ca(2+)](i) 后 48 小时，与模拟转染对照相比，沉默后 CCSM 细胞中的 ICW 幅度和扩散没有显着差异，而短期阻断显着增加了基础 [Ca(2+)](i) 水平并放大了 CCSM 细胞中的 Ca(2+) 信号传导。微阵列分析表明Ca(2+)稳态和平滑肌张力调节途径中的几个基因的表达发生了显着改变。结论。我们的结果表明，虽然短期阻断 MaxiK 通道与基础 [Ca(2+)](i) 的增加有关，但 Ca(2+) 稳态在沉默后 48 小时内恢复。我们假设调节 [Ca(2+)](i) 和 CCSM 音调的不同途径通过分子串扰联系在一起，并且它们的协调调节是旨在维持 Ca(2+) 稳态和 CCSM 音调的补偿机制的一部分。 Calenda G、Suadicani SO、Iglesias R、Spray DC、Melman A 和 Davies KP。沉默下体平滑肌细胞中的 MaxiK 活性会启动维持钙稳态的补偿机制。 《性医学杂志》2011 年；8：2191-2204。
Introduction. The MaxiK potassium channel is regulated by voltage and intracellular calcium, and plays a critical role in regulating intracellular calcium concentration ([Ca(2+)](i)), which is the ultimate determinant of smooth muscle tone. Tight control of corpus cavernosum smooth muscle (CCSM) tone is critically important and misregulation can result in erectile dysfunction.Aim. Because of the tight functional linkage of MaxiK and calcium channel activity, the aim of this study was to determine the effects of silencing and pharmacological inhibition of MaxiK on calcium homeostasis and intercellular calcium signaling in CCSM cells.Methods. We compared changes in the basal intracellular [Ca(2+)](i) and parameters defining intercellular calcium wave (ICW) spread in 48 hours MaxiK silenced CCSM cells vs. acute blockade of the channel with iberiotoxin. To analyze changes occurring in gene expression we performed micro-array analysis following MaxiK silencing for 48 hours.Main Outcome Measures. Changes in Fura-2 fluorescence intensities were measured to evaluate basal [Ca(2+)](i) levels and ICW parameters. Microarray analysis of global gene expression was performed.Results. Forty-eight hours after MaxiK silencing the basal [Ca(2+)](i), the ICW amplitude and spread among CCSM cells were not markedly different in silenced compared to mock transfected controls, whereas short-term blockade significantly increased basal [Ca(2+)](i) level and amplified Ca(2+) signaling among CCSM cells. Micro-array analysis showed that several genes within Ca(2+) homeostasis and smooth muscle tone regulation pathways had significantly altered expression.Conclusions. Our results indicate that while short-term blockade of the MaxiK channel is associated with an increase in basal [Ca(2+)](i), Ca(2+) homeostasis is restored during the 48 hours period following silencing. We hypothesize that the different pathways regulating [Ca(2+)](i) and CCSM tone are linked through molecular crosstalk and that their coordinated regulation is part of a compensatory mechanism aimed to maintain Ca(2+) homeostasis and CCSM tone. Calenda G, Suadicani SO, Iglesias R, and Spray DC, Melman A, and Davies KP. Silencing MaxiK activity in corporal smooth muscle cells initiates compensatory mechanisms to maintain calcium homeostasis. J Sex Med 2011;8:2191-2204.