Alvarez Osvaldo, Castillo Karen, Carmona Emerson, Gonzalez Carlos, Latorre Ramon
Departamento de Biología, Facultad de Ciencias, Universidad de Chile, Santiago, Chile.
Facultad de Ciencias, Centro Interdisciplinario de Neurociencia de Valparaíso, Universidad de Valparaíso, Valparaíso, Chile.
Methods Mol Biol. 2019;1987:167-185. doi: 10.1007/978-1-4939-9446-5_11.
A complete characterization of temperature -and voltage-activated TRP channel gating requires a precise determination of the absolute probability of opening in a wide range of voltages, temperatures, and agonist concentrations. We have achieved this in the case of the TRPM8 channel expressed in Xenopus laevis oocytes. Measurements covered an extensive range of probabilities and unprecedented applied voltages up to 500 mV. In this chapter, we describe animal care protocols of patch-clamp pipette preparation, temperature control methods, and analysis of ionic currents to obtain reliable absolute open channel probabilities.
对温度和电压激活的瞬时受体电位（TRP）通道门控进行完整表征，需要精确测定在广泛的电压、温度和激动剂浓度范围内通道开放的绝对概率。我们在非洲爪蟾卵母细胞中表达的TRPM8通道的研究中实现了这一点。测量涵盖了广泛的概率范围以及高达500 mV的前所未有的施加电压。在本章中，我们描述了膜片钳吸管制备的动物护理方案、温度控制方法以及离子电流分析，以获得可靠的绝对开放通道概率。
