Prostaglandin F2alpha receptor (FP) expression in human umbilical vein (HUV).

CESIO C, PELOROSSO F, ROTHLIN R, ERRASTI, A.

Ciudad de Córdoba

Congreso; XXXVIII Reunion Anual de la Asociacion Argentina de Farmacologia Experimental.; 2006

Asociación Argentina de Farmacología Experimental (SAFE)

Prostaglandin F2a receptor (FP) expression in human umbilical vein (HUV). Cesio, C, Pelorosso F, Rothlin, R Errasti, A. III Cátedra de Farmacología. Facultad de Medicina. UBA. Paraguay 2155, piso 9, 1121. farmaco3@fmed.uba.ar. Introduction: Prostaglandin F2a (PGF2a) and its analogs bimatoprost free acid and latanoprost free acid (selective FP receptor agonists), promote constriction of HUV. This effect is selectively blocked by FP receptor antagonist, AL-8810, suggesting that FP receptors are involved in the vasoconstrictor effect. Therefore, the aim of the present study was to analize the presence of FP receptor at mRNA and protein level in HUV by RT-PCR and Western blot, respectively. Methods and results: Total RNA (HUV) and proteins (HUV and mouse uterus) were extracted employing Trizol and RIPA lysis buffer, respectively. RNA was quantified at 260/280nm and proteins were measured with Bradford at 595 nm. PCR products were electrophoresed on 2% agarose gels with ethidium bromide and photographed under UV. Endonuclease digestion was used to confirm product identity. Proteins were electrophoresed on 10% SDS-PAGE and electrotransferred to PVDF membranes which were blocked in TTBS buffer with 5% milk; then incubated overnight with anti rabbit FP receptor (murine) polyclonal antibodies. Membranes were revealed with alkaline phosphatase-conjugated goat anti-rabbit IgG. Immunoreactive bands were detected by chemiluminescence and compared with those obtained in mouse uterus (control tissue). Conclusion: The results indicate that whole HUV express a FP receptor at mRNA level and a protein of similar molecular weight that one observed in mouse uterus, a rich source of FP receptors.