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Biology
电生理测量在中性转运体活性研究中的应用
电生理测量在中性转运体活性研究中的应用
JoVE Journal
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JoVE Journal Biology
Application of Electrophysiology Measurement to Study the Activity of Electro-Neutral Transporters

电生理测量在中性转运体活性研究中的应用

Full Text
7,524 Views
11:51 min
February 3, 2018

DOI: 10.3791/56630-v

Victor Babich1,2, Matthew K. Henry1, Francesca Di Sole1

1Physiology and Pharmacology Department,Des Moines University, 2School of Liberal Arts and Sciences,Mercy College of Health Sciences

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Please note that some of the translations on this page are AI generated. Click here for the English version.

本文介绍了质子选择性电极和膜片夹紧方法在质子传输系统中的应用。这些方法克服了一般用于研究质子转运活动的一些技术限制, 如灵敏度适中、时间分辨力和细胞内环境控制不足。

巨型贴片与离子选择性微电极相结合的方法是一种用于检测和定量离子通量的通用方法。具体来说,它用于测量由电中性离子转运蛋白、钠氢交换剂或 NHE 的活动产生的质子通量,该质子通量在细胞膜上表达。钠-氢交换器在质膜附近产生质子梯度,此处表示为橙色环。

通过全细胞膜片钳记录结合使用质子选择性微电极的技术来测量跨细胞膜的质子梯度。质子选择性微电极靠近电池表面移动,在那里它记录从电池中移出的质子的通量,如图所示的位置 A.远离电池表面,在那里它记录位置 B 指示的溶液中存在的游离质子浓度,然后,回到位置 A,靠近细胞膜, 在重复的振荡运动中。记录从 A 到 B 的电压差,并取决于 NHE 活性的代表。

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