Electrochemical measurement of endogenous serotonin release from human blood platelets.
Electrochemical measurement of endogenous serotonin release from human blood platelets.
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DOI：
10.1021/ac102929y
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发表时间：
2011-04-01
影响因子：
7.4
通讯作者：
Haynes, Christy L.
中科院分区：
文献类型：
--
作者：
Ge, Shencheng;Woo, Emily;White, James G.;Haynes, Christy L.
血流中的血小板聚集与血小板的分泌功能密切相关，其基础是几种类型的细胞质分泌颗粒，每一种都隔离不同的化学信使种类。致密体颗粒是一种重要的分泌颗粒，负责存储小分子化学信使。当血小板被激活时，这些小分子的及时和快速释放对于促进血小板聚集至关重要。因此，需要能够实时测量颗粒内容物释放的技术来了解血小板分泌和聚集的基本特性。现有技术缺乏足够的时间分辨率或需要潜在有毒的外源性试剂来实时测量颗粒含量释放。在这里，我们展示了一种基于内源性电活性化学信使5-羟色胺(5-羟色胺或5-羟色胺)的无标记电化学方法，用于实时测量血小板悬浮液中致密体颗粒的分泌；使用碳纤维微电极的快速扫描循环伏安法(FSCV)因其优异的时间分辨率、高灵敏度以及能够为分子鉴定提供电化学签名循环伏安图而被选择。成功地测量了凝血酶刺激的人血小板悬液中5-羟色胺的实时释放，发现大量5-羟色胺的释放量和时间过程与单个血小板的测量数据很好地吻合，从而证实了颗粒分泌的准确特征。此外，该电化学方法被应用于研究血小板的刺激-分泌偶联，5-羟色胺的储存和释放动力学，以及Hermansky-Pudlak综合征(HPS)血小板的化学信使储存缺陷，清楚地展示了该方法在揭示正常和疾病血小板的分泌行为方面的潜力。
Platelet aggregation in the blood stream is tightly associated with the secretory function of platelets based on several types of cytoplasmic secretory granules, each sequestering distinct chemical messenger species. Dense-body granules are one prominent type of secretory granules responsible for storing small molecule chemical messengers. Upon platelet activation, the timely and rapid release of these small molecules is critical in facilitating platelet aggregation. Therefore, techniques capable of measuring real-time granule content release are needed to understand the fundamental properties of platelet secretion and aggregation. Existing techniques lack adequate time resolution or require potentially toxic exogenous reagents for real-time measurement of granule content release. Herein, we demonstrate a label-free electrochemical method based on the endogenous electroactive chemical messenger serotonin (5-hydroxytryptamine or 5-HT) for the real-time measurement of dense-body granule secretion from platelet suspensions; fast-scan cyclic voltammetry (FSCV) using carbon-fiber microelectrodes was chosen based on its excellent temporal resolution, high sensitivity, and the ability to provide the electrochemical signature cyclic voltammograms for molecular identification. Real-time serotonin release from thrombin-stimulated human platelet suspensions was successfully measured and the amount and time course of the bulk serotonin release were found to agree well with data obtained from single platelet measurements, thus confirming accurate characterization of granular secretion. Furthermore, this electrochemical method was applied to study the stimulation-secretion coupling in platelets, serotonin storage and release dynamics with applied pharmacological agents, and chemical messenger storage deficiency in Hermansky-Pudlak Syndrome (HPS) platelets, and has clearly demonstrated the potential of this method to reveal secretion behavior in both normal and diseased platelets.
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影响因子：
2.9
作者：
ANDERSON, GM;HALL, LM;COHEN, DJ
通讯作者：
COHEN, DJ
影响因子：
7.4
作者：
Ge, Shencheng;White, James G.;Haynes, Christy L.
通讯作者：
Haynes, Christy L.
DOI：
10.1016/0304-4165(85)90202-8
发表时间：
1985-01-01
期刊：
BIOCHIMICA ET BIOPHYSICA ACTA
影响因子：
--
作者：
MCCREA, JM;ROBINSON, P;GERRARD, JM
通讯作者：
GERRARD, JM
影响因子：
2.9
作者：
Ge, Shencheng;Wittenberg, Nathan J.;Haynes, Christy L.
通讯作者：
Haynes, Christy L.
影响因子：
7.4
作者：
CHEN, TK;LUO, GO;EWING, AG
通讯作者：
EWING, AG