Toth Adam V, Bartok Adam
Department of Biochemistry, Semmelweis University, Budapest, Hungary.
HCEMM-SE Molecular Channelopathies Research Group, Budapest, Hungary.
Methods Mol Biol. 2025;2908:163-170. doi: 10.1007/978-1-0716-4434-8_11.
The transient receptor potential (TRP) family of ion channels contains a large array of receptors of pain, taste, or temperature. Thus far, several TRP channels have been confirmed as warm or cold sensors; however, the exact molecular mechanism of heat detection remains obscure. The methods of investigation of temperature sensitivity involve several different techniques, e.g., in vivo experiments, fluorescence-based Ca measurements, and electrophysiology. Here we describe an electrophysiological configuration that allows the direct assessment of the temperature sensitivity of the TRPM2 channel by measuring the temperature dependence of microscopic and macroscopic currents, apparent ligand binding affinities, and the gating parameters of the channel under the precise control of temperature and activating ligand concentrations. The technique described here may be adapted to explore temperature sensitive behavior of various other ion channels.
瞬时受体电位（TRP）离子通道家族包含大量与疼痛、味觉或温度相关的受体。到目前为止，已有几种TRP通道被确认为温觉或冷觉感受器；然而，热觉检测的确切分子机制仍不清楚。研究温度敏感性的方法涉及几种不同的技术，例如体内实验、基于荧光的钙离子测量以及电生理学。在此，我们描述一种电生理配置，通过在温度和激活配体浓度的精确控制下测量微观和宏观电流的温度依赖性、表观配体结合亲和力以及通道的门控参数，从而直接评估TRPM2通道的温度敏感性。这里描述的技术可用于探索各种其他离子通道的温度敏感行为。
