engleski

Examining the activation status of OCRL inhibited platelets by flow cytometry

We researched the involvement of phosphoinositides in the development and function of the smallest blood cells, platelets. In particular, I investigate phosphatidylinositol-4-monophosphate (PI4P), phosphatidylinositol-4, 5-bisphosphate [PI(4, 5)P2], and the oculocerebrorenal syndrome of Lowe protein (OCRL) that dephosphorylates PI(4, 5)P2 to PI4P. Mutations in OCRL cause Lowe syndrome, which is characterized by a triade of symptoms affecting the brain, the eyes, and the kidneys, but it has also been shown that these patients have problems with bleedings suggesting platelet (PLT) disfunction. We started our research by inhibiting OCRL and examining the spreading ability of human and mouse PLTs by immunocytochemistry, then we moved on to researching signalling pathways in human PLTs that OCRL might be affecting and we preliminary characterized megakaryocytes and PLTs from OCRL KO mice by several different methods. The next step was to analyze how OCRL affects PLT function upon activation with different agonists by flow cytometry. We started by inhibiting PLTs for OCRL and activating them with a thrombin mimetic, staining for two PLT activation markers and examining them by flow cytometry.

OCRL, platelets, p-selectin, integrin, flow cytometry

nije evidentirano