1. Introduction
Amperometry is arguably the most widely used technique for studying the exocytosis of biological amines, allowing the study of the dynamics and kinetics of this cellular mechanism in real time. In recent decades, multielectrode array (MEA) devices have been developed and increase the efficiency and speed of amperometric measurements. Platelets are the most accessible cells to study the exocytosis of amines as they have 90% of blood serotonin included in their granules, which is avidly uptaken from the blood and released by exocytosis. Furthermore, platelets are not able to synthesize this amine.
2. Methods
We have optimized boron-doped diamond (BDD) MEA systems that allow the detection of amperometric recordings from the quantum release of serotonin from human platelets. With this technique, exocytotic release phenomena are recorded as a succession of discrete signals in the form of spikes that result from electric current intensities measured during the oxidation of the released serotonin. Our initial results were carried out with 16 microelectrode devices on silicon substrate (BDD-on-silicon MEA) [1]. In this conference, we present transparent MEA devices with 16 microelectrodes that allow microscopy observation too.
3. Results
BDD-on-quartz MEA. BDD-on-quartz MEA devices exhibit the same excellent electrochemical properties as BDD-on-silicon MEA: we present a comparative study of quantum and kinetics data obtained with both MEA chips from unloaded platelets and after loading the platelets with 10 µM serotonin for 2 h. And finally, we show examples of the different types of spikes detected.
4. Conclusions
We demonstrate the effectiveness of BDD-on-quartz MEA devices for amperometric studies of serotonin exocytosis from human platelets.
Author Contributions
Conceptualization, R.B. and A.P.; methodology, R.G.-B., P.M. and A.M.; formal analysis, R.G.-B., P.M. and A.M.; software and validation, R.E.S.; review and editing, R.G.-B., R.B. and AP. All authors have read and agreed to the published version of the manuscript.
Funding
This work was partially supported by the Spanish Ministry of Ciencia e Innovación, grant No. PID2020-116589GB-I00, to R.B.
Institutional Review Board Statement
This study was conducted in accordance with the Declaration of Helsinki and was approved by the Ethical Committee at the University of La Laguna (CEIBA2017-0244) and by the Ethical Committee at the Canary Islands Health Services (CHUC_2020_80) for the use of human samples.
Informed Consent Statement
Informed consent was obtained from all subjects involved in the study.
Data Availability Statement
The data presented in this study are available in the references.
Conflicts of Interest
The authors declare no conflict of interest.
Reference
- González Brito, R.; Montenegro, P.; Méndez, A.; Carabelli, V.; Tomagra, G.; Shabgahi, R.E.; Pasquarelli, A.; Borges, R. Multielectrode Arrays as a Means to Study Exocytosis in Human Platelets. Biosensors 2023, 13, 86. [Google Scholar] [CrossRef] [PubMed]
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