A Turbidimetric Analysis to Monitor Collagen-Induced Aggregation of Pretreated Human Platelets
Transcription
Begin with glass cuvettes containing pre-treated, human platelet suspension containing non-activated platelets and fibrinogens — a plasma glycoprotein.
The cuvettes include a stirring device for uniform mixing.
Place the control cuvette in an aggregometer that measures the aggregation by assessing turbidity.
Introduce collagen — an agonist for platelet aggregation, interacting with platelet's surface receptors and activating downstream signaling cascades.
This results in ATP release via the Pannexin 1 channel — a platelet surface channel, initiating platelet activation and shape change, leading to a depression in the aggregation curve.
Fibrinogen further crosslinks activated platelets, promoting collagen-induced platelet aggregation, altering turbidity, and causing a gradual rise in the aggregation curve.
Treat the test cuvette with a high concentration of an inhibitor of the Pannexin 1 channel that blocks this channel.
Add collagen to the cuvette. Inhibitor interactions prevent ATP release and hinder collagen-induced platelet aggregation, evident from unchanged turbidity and a flat aggregation curve.
