Platelets isolation and ectonucleotidase assay: Revealing functional aspects of the communication between the vasculature and the immune system.

Publication date: Oct 01, 2024

Platelets are enucleated fragments of cells with a diversity of internal granules. They are responsible for functions related to hemostasis, coagulation, and inflammation. The activation of these processes depends on a cascade coordinated by cytokines, chemokines, and components of purinergic signaling, such as ATP, ADP, and adenosine. Platelets express distinct components of the purinergic system: P2X1, P2Y1, PY12, and P2Y14 receptors; and the ectonucleotidases NTPDase, NPP, and 5NTE (ecto-5′-nucleotidase). Except for P2Y14, which has not yet exhibited a known function, all other components relate to the biological processes mentioned before. Platelets are known to display specific responses to microorganisms, being capable of recognizing pathogen-associated molecular patterns (PAMPs), engulfing certain classes of viruses, and participating in NETosis. Platelet function dysregulation implicates various pathophysiological processes, including cardiovascular diseases (CVDs) and infections. In COVID-19 patients, platelets exhibit altered purinergic signaling and increased activation, contributing to inflammation. Excessive platelet activation can lead to complications from thrombosis, which can affect the circulation of vital organs. Therefore, controlling the activation is necessary to end the inflammatory process and restore homeostasis. Ectonucleotidases, capable of hydrolyzing ATP, ADP, and AMP, are of fundamental importance in activating platelets, promising pharmacological targets for clinical use as cardiovascular protective drugs. In this review, we revisit platelet biology, the purinergic receptors and ectonucleotidases on their surface, and their importance in platelet activity. Additionally, we describe methods for isolating platelets in humans and murine, as well as the main techniques for detecting the activity of ectonucleotidases in platelets. Considering the multitude of functions revealed by platelets and their potential use as potent bioreactors able to secrete and present molecules involved in the communication of the vasculature with the immune system, it is crucial to deeply understand platelet biology and purinergic signaling participation to contribute to the developing of therapeutic strategies in diseases of the cardiovascular, inflammatory, and immune systems.

Concepts Keywords
Biology 5′-Nucleotidase
Ectonucleotidase 5′-Nucleotidase
Homeostasis Adenosine Triphosphatases
Pathophysiological Adenosine Triphosphatases
Viruses Animals
Blood Platelets
Cell Separation
COVID-19
Ectonucleotidases
Humans
Inflammation
Platelet Activation
Platelets isolation techniques
Purinergic system
SARS-CoV-2
Signal Transduction
Thrombocytes

Semantics

Type Source Name
disease IDO assay
pathway REACTOME Immune System
pathway REACTOME Hemostasis
disease MESH inflammation
drug DRUGBANK ATP
drug DRUGBANK Adenosine
drug DRUGBANK Nandrolone phenpropionate
disease MESH cardiovascular diseases
disease MESH infections
disease MESH COVID-19
pathway KEGG Platelet activation
disease MESH complications
disease MESH thrombosis
disease IDO process
disease IDO blood
disease IDO cell
pathway REACTOME Signal Transduction

Original Article

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