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首页> 外文期刊>Journal of Neuroscience Methods >Rotating disk electrode voltammetric measurements of serotonin transporter kinetics in synaptosomes.
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Rotating disk electrode voltammetric measurements of serotonin transporter kinetics in synaptosomes.

机译：旋转盘电极伏安法测定突触小体中5-羟色胺转运蛋白的动力学。

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Altered serotonin (5-HT) signaling is implicated in several neuropsychiatric disorders, including depression, anxiety, obsessive-compulsive disorder, and autism. The 5-HT transporter (SERT) modulates 5-HT neurotransmission strength and duration. This is the first study using rotating disk electrode voltammetry (RDEV) to measure 5-HT clearance. SERT kinetics were measured in whole brain synaptosomes. Uptake kinetics of exogenous 5-HT were measured using glassy carbon electrodes rotated in 500 muL glass chambers containing synaptosomes from SERT-knockout (-/-), heterozygous (+/-), or wild-type (+/+) mice. RDEV detected 5-HT concentrations of 5nM and higher. Initial velocities were kinetically resolved with K(m) and V(max) values of 99+/-35 standard error of regression (SER) nM and 181+/-11 SER fmol/(sxmg protein), respectively in wild-type synaptosomes. The method enables control over drug and chemical concentrations, facilitating interpretation of results. Results are compared in detail to other techniques used to measure SERT kinetics, including tritium labeled assays, chronoamperometry, and fast scan cyclic voltammetry. RDEV exhibits decreased 5-HT detection limits, decreased vulnerability to 5-HT oxidation products that reduce electrode sensitivity, and also overcomes diffusion limitations via forced convection by providing a continuous, kinetically resolved signal. Finally, RDEV distinguishes functional differences between genotypes, notably, between wild-type and heterozygous mice, an experimental problem with other experimental approaches.
机译：血清素（5-HT）信号改变与多种神经精神疾病有关，包括抑郁症，焦虑症，强迫症和孤独症。 5-HT转运蛋白（SERT）调节5-HT神经传递强度和持续时间。这是首次使用转盘电极伏安法（RDEV）测量5-HT清除率的研究。在全脑突触小体中测量了SERT动力学。使用在500μL玻璃室中旋转的玻璃碳电极测量外源5-HT的吸收动力学，该玻璃室包含来自SERT基因敲除（-/-），杂合（+/-）或野生型（+ / +）小鼠的突触小体。 RDEV检测到5-HT浓度为5nM及更高。分别用野生型突触体中的K（m）和V（max）值分别以99 +/- 35标准回归误差（SER）nM和181 +/- 11 SER fmol /（sxmg蛋白）动力学解析初始速度。该方法可以控制药物和化学药品的浓度，从而有助于结果的解释。将结果与其他用于测量SERT动力学的技术进行了详细比较，包括tri标记的测定，计时电流法和快速扫描循环伏安法。 RDEV显示出降低的5-HT检测极限，降低了对5-HT氧化产物的敏感性，从而降低了电极的灵敏度，并且通过提供连续的，动态分辨的信号，通过强制对流克服了扩散限制。最后，RDEV可以区分基因型之间的功能差异，尤其是野生型和杂合型小鼠之间的功能差异，这是其他实验方法的实验问题。

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