INVESTIGADORES
SZPILBARG Natalia
congresos y reuniones científicas
Título:
AQP3 and AQP9 expression in extracellular microvesicle-enriched samples of plasma and supernatant of placental explants from healthy pregnant women at term
Autor/es:
SZPILBARG, NATALIA; COROMINAS, ANA; FERREIRÓS, ALBERTO; RUIZ, JULIETA; CASALE, ROBERTO; DAMIANO, ALICIA ERMELINDA
Lugar:
Bogotá
Reunión:
Congreso; IX SLIMP VIRTUAL MEETING; 2022
Institución organizadora:
Sociedad Latinoamericana de Interacción Materno-Fetal y Placenta ? Latin American Society for Materno Fetal Interaction and Placenta
Resumen:
Aim: To detect AQP3 and AQP9 protein expression in extracellular microvesicle-enrichedsamples of plasma and supernatant of placental explants from healthy pregnant womenat term.Methodology: EDTA anticoagulated blood samples and placentas were obtained from 6women with healthy term pregnancies, under Ethics Committee protocol and signedinformed consent at Hospital Posadas in Buenos Aires. Explants were prepared andcultured for 24 hours under standard culture conditions. Plasmas and the correspondingsupernatants of placental explants from each patient were subjected to a protocol ofdifferential centrifugation, filtering, and ultracentrifugation to obtain samples enrichedin extracellular microvesicles. Each sample was analyzed by Dynamic light scattering(DLS) in a Zetasizer Nano-ZSP equipment (Malvern Instruments, UK) equipped with a HeNe laser (633nm) and a digital correlator, model ZEN5600, to determine the size of themicrovesicles. Subsequently, the samples were analyzed by Western blot to determinethe expression of AQP3, AQP9, and PLAP using specific antibodies: anti-PLAP HD 11F7(1:200, Santa Cruz), anti AQP3 (1:1000, Alpha diagnostics), anti- AQP9 (1:1000, Alphadiagnostics) and secondary antibodies (Jackson), using homogenates from each placentaas positive controls.Results: One or two (depending on the case) predominant microvesicle populations,whose size ranged between 40 and 220nm, were obtained in plasmas and theircorresponding supernatants of placental explant cultures. Using Western blot, thepresence of PLAP (syncytiotrophoblast marker), AQP3 and AQP9 was evidenced in allthe samples analyzed.Conclusions: These preliminary results allow us to lay the foundations to be able, in thefuture, to determine the presence of AQPs in microvesicles of placental origin in theplasma of pregnant women at any stage of pregnancy. In this sense, monitoring theexpression levels of these proteins in healthy and pathological pregnancies may also letus learn more about their functions during pregnancy.