Abstract
This paper introduces a new technique combining microdialysis with patch clamp to detect the changes in ATP (adenosine triphosphate) concentration in acupoints during acupuncture. The microdialysis probe was implanted into the Zusanli acupoint (ST 36) of adult SD (Sprague Dawley) rats to sample acupoint fluid containing ATP released during acupuncture. Then, the fluid with ATP was delivered in real time to 293 T cells overexpressing P2X3 receptors, with which we could carry out patch clamp experiments. The results showed that changes in membrane currents could reflect changes in the concentration of ATP. Thus, we can successfully detect ATP released in acupoints during acupuncture in real time. This technique provides us with a new way to study the mechanism of acupuncture signal initiation.
Highlights
Microdialysis has been widely used in life sciences
Different concentrations of Adenosine triphosphate (ATP) can be detected by the system in real time in vitro First, we performed in vitro experiments with different concentrations of ATP standard solutions (10 and 100 μM). 293 T cells overexpressing P2X3 receptors (293TX for short) were used for the patch clamp recording
As ATP was dialyzed into the chamber, the membrane currents changed
Summary
Compared to other sampling techniques, microdialysis has distinctive advantages. The sampling process can be continuous for hours or even days without major changes in the microenvironment. Macromolecules, such as proteins, are excluded from the semipermeable membrane so that the samples can be directly used for subsequent analysis without a cumbersome purifying procedure [18]. Due to its ability for high temporal resolution recording of ion channel currents from cells or cell-free membrane patches, the patch clamp technique has become the primary method for studying cell signal transduction mechanisms. Different types of ion channels have been studied, such as voltage-gated K+ channels [2], ligand-
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