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Development and Characterization of In Vitro Microvessel Network and Quantitative Measurements of Endothelial [Ca2+]i and Nitric Oxide Production
Published on: May 19, 2016
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Simultaneous analysis of vascular norepinephrine and ATP release using an integrated microfluidic system
Alexandra D Townsend1, Gerald H Wilken2, Kyle K Mitchell3
1Department of Chemistry, Saint Louis University, St. Louis, MO 63103, United States.
Journal of Neuroscience Methods
|March 27, 2016
Summary
Researchers developed a new method using 3-D printing to simultaneously detect norepinephrine and adenosine triphosphate (ATP) from rat mesenteric arteries in near real-time, improving upon older techniques.
Area of Science:
- Physiology
- Neuroscience
- Analytical Chemistry
Background:
- Sympathetic nerves release norepinephrine, ATP, and neuropeptide Y, influencing vascular tone.
- This study investigates the co-release of norepinephrine and ATP from rat mesenteric arterial sympathetic nerves.
Purpose of the Study:
- To develop and validate a novel technique for simultaneous, near-real-time detection of norepinephrine and ATP.
- To compare this new method with existing techniques like HPLC for neurotransmitter analysis.
Main Methods:
- A 3-D printing-based system was developed to integrate perfusate delivery with simultaneous detection methods.
- Norepinephrine was detected using microchip-based amperometry, and ATP was detected via on-line chemiluminescence.
- The method was applied to an isolated perfused mesenteric arterial bed of the rat.
Main Results:
- The technique successfully achieved simultaneous detection of norepinephrine and ATP in near real-time.
- Stimulated norepinephrine levels reached 363nM, and ATP levels reached 125nM.
- The limit of detection for norepinephrine was 80nM, and for ATP was 35nM.
Conclusions:
- The described method enables successful simultaneous and near-real-time detection of norepinephrine and ATP.
- This advancement offers an improvement over traditional HPLC methods, which involve sample derivatization and delays, potentially leading to degradation.
