Measurement of ATP-Induced Membrane Potential Changes in IVD cells.
Measurement of ATP-Induced Membrane Potential Changes in IVD cells.
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DOI：
10.1007/s12195-014-0355-6
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发表时间：
2014-12-01
影响因子：
2.8
通讯作者：
Huang, Chun-Yuh Charles
中科院分区：
文献类型：
--
作者：
Gonzales, Silvia;Rodriguez, Brittany;Barrera, Carlos;Huang, Chun-Yuh Charles
细胞外腺苷-5'-三磷酸（ATP）在多种细胞和组织中引发生物反应，并激活影响细胞膜电位和兴奋性的信号级联反应。压缩载荷促进了椎间盘（IVD）细胞ATP的产生和释放，而高水平的细胞外ATP在IVD的髓核（NP）中积累。本研究利用电位敏感染料二-8-丁基-氨基-萘-乙烯-吡啶-丙基磺酸盐（di-8-ANEPPS），建立了一种无创系统来测量atp诱导的猪IVD细胞膜电位变化。在单层和三维培养中观察NP和纤维环细胞对ATP的反应。细胞外ATP诱导IVD细胞膜电位变化的模式和幅度与细胞类型、培养条件和ATP剂量有关。此外，P2X4嘌呤能受体基因在两种细胞类型中均有表达。piridoxalphosphate -6-azophenyl-2', 4'- diulfonate （PPADS）是一种P2受体的非竞争性抑制剂，可以抑制ATP诱导的反应，这表明ATP可能通过P2嘌呤能受体调节IVD细胞的生物活性。
Extracellular adenosine-5'-triphosphate (ATP) triggers biological responses in a wide variety of cells and tissues and activates signaling cascades that affect cell membrane potential and excitability. It has been demonstrated that compressive loading promotes ATP production and release by intervertebral disc (IVD) cells, while a high level of extracellular ATP accumulates in the nucleus pulposus (NP) of the IVD. In this study, a noninvasive system was developed to measure ATP-induced changes in the membrane potential of porcine IVD cells using the potential sensitive dye di-8-butyl-amino-naphthyl-ethylene-pyridinium-propyl-sulfonate (di-8-ANEPPS).The responses of NP and annulus fibrosus (AF) cells to ATP were examined in monolayer and 3-dimensional cultures. It was found that the pattern and magnitude of membrane potential change in IVD cells induced by extracellular ATP depended on cell type, culture condition, and ATP dose. In addition, gene expression of P2X4 purinergic receptor was found in both cell types. Inhibition of the ATP-induced response by pyridoxalphosphate-6-azophenyl-2', 4'-disulfonate (PPADS), a non-competitive inhibitor of P2 receptors, suggests that ATP may modulate the biological activities of IVD cells via P2 purinergic receptors.
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影响因子：
2.8
作者：
Fernando, Hanan N.;Czamanski, Jessica;Yuan, Tai-Yi;Gu, Weiyong;Salahadin, Abdi;Huang, Chun-Yuh Charles
通讯作者：
Huang, Chun-Yuh Charles
影响因子：
3
作者：
BUCKWALTER, JA
通讯作者：
BUCKWALTER, JA
DOI：
10.4161/cc.8.21.9888
发表时间：
2009-11-01
期刊：
Cell cycle (Georgetown, Tex.)
影响因子：
--
作者：
Blackiston DJ;McLaughlin KA;Levin M
通讯作者：
Levin M
影响因子：
64.5
作者：
Levin, M;Thorlin, T;Mercola, M
通讯作者：
Mercola, M
影响因子：
2.4
作者：
DiFranco, M;Capote, J;Vergara, JL
通讯作者：
Vergara, JL