Schapira Isabella, O'Neill Margaret R, Russo-Savage Lillian, Narla Terdha, Laprade Kathryn A, Stafford James M, Ou Yangguang
Department of Chemistry, University of Vermont USA
Neuroscience Graduate Program, University of Vermont USA.
RSC Adv. 2023 Sep 4;13(37):26203-26212. doi: 10.1039/d3ra04551j. eCollection 2023 Aug 29.
Despite the fact that tryptophan (Trp) is an essential amino acid that humans typically obtain through diet, there are several interesting tryptophan dynamics at play in the body. Quantifying and understanding these dynamics are crucial in studies of depression, autism spectrum disorder, and other disorders that involve neurotransmitters directly synthesized from tryptophan. Here we detail the optimization of waveform parameters in fast scan cyclic voltammetry at carbon fiber microelectrodes to yield four-fold higher sensitivity and six-fold higher selectivity compared to previously reported methods. We demonstrate the utility of our method in measuring (1) exogenous Trp dynamics from administration of Trp to PC-12 cells with and without overexpression of tryptophan hydroxylase-2 and (2) endogenous Trp dynamics in pinealocyte cultures with and without stimulation norepinephrine. We observed interesting differences in Trp dynamics in both model systems, which demonstrate that our method is indeed sensitive to Trp dynamics in different applications.
尽管色氨酸（Trp）是一种人体通常通过饮食获取的必需氨基酸，但人体内存在着几种有趣的色氨酸动态变化。在抑郁症、自闭症谱系障碍以及其他直接涉及由色氨酸合成的神经递质的疾病研究中，量化和理解这些动态变化至关重要。在此，我们详细介绍了碳纤维微电极快速扫描循环伏安法中波形参数的优化，与先前报道的方法相比，该方法灵敏度提高了四倍，选择性提高了六倍。我们展示了该方法在测量以下两方面的效用：（1）在有和没有过表达色氨酸羟化酶-2的情况下，向PC-12细胞施用色氨酸后的外源性色氨酸动态变化；（2）在有和没有去甲肾上腺素刺激的情况下，松果体细胞培养物中的内源性色氨酸动态变化。我们在这两个模型系统中均观察到色氨酸动态变化存在有趣的差异，这表明我们的方法在不同应用中确实对色氨酸动态变化敏感。
