Abstract
Determinations of platelet receptor functions are indispensable diagnostic indicators of cardiovascular and hemostatic diseases including hereditary and acquired receptor defects and receptor responses to drugs. However, presently available techniques for assessing platelet function have some disadvantages, such as low sensitivity and the requirement of large sample sizes and unphysiologically high agonist concentrations. Our goal was to develop and initially characterize a new technique designed to quantitatively analyze platelet receptor activation and platelet function on the basis of measuring changes in low angle light scattering. We developed a novel technique based on low angle light scattering registering changes in light scattering at a range of different angles in platelet suspensions during activation. The method proved to be highly sensitive for simultaneous real time detection of changes in size and shape of platelets during activation. Unlike commonly-used methods, the light scattering method could detect platelet shape change and aggregation in response to nanomolar concentrations of extracellular nucleotides. Furthermore, our results demonstrate that the advantages of the light scattering method make it a choice method for platelet receptor monitoring and for investigation of both murine and human platelets in disease models. Our data demonstrate the suitability and superiority of this new low angle light scattering method for comprehensive analyses of platelet receptors and functions. This highly sensitive, quantitative, and online detection of essential physiological, pathophysiological and pharmacological-response properties of human and mouse platelets is a significant improvement over conventional techniques.
Highlights
Platelet activation and aggregation play key roles in both normal hemostasis and pathological bleeding and thrombosis
Our results demonstrate that the advantages of the light scattering method make it a choice method for platelet receptor monitoring and for investigation of both murine and human platelets in disease models
The setup used here provides a simultaneous observation of the intensity of the scattered light at angles ranging from 1° to 12° (Supplementary Figure 2)
Summary
Platelet activation and aggregation play key roles in both normal hemostasis and pathological bleeding and thrombosis. LTA and even other recently developed aggregometry tests have obvious limitations, such as low sensitivity, considerable sample consumption and the required unphysiologically high concentrations of stimulant. Since the introduction of LTA numerous attempts have been made to improve the sensitivity of this technique These approaches, require a complex setup and/or sophisticated analysis and have not been fully established as functional platelet tests [9, 10]. A considerable improvement has been achieved by switching from detection by light transmittance to an electrical detection by measuring the impedance of the sample [11] Even these novel techniques are still hampered by the clearly unphysiologically high concentration of stimulants required to achieve platelet aggregation and by their inability to detect platelet shape changes. Unlike commonly-used methods, the Keywords: aggregation; platelet; P2X1 receptor; P2Y receptors
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