INVESTIGADORES
ROLDAN OLARTE Eugenia Mariela
congresos y reuniones científicas
Título:
“Proteolytic activity and gene expression of plasminogen activator in porcine oviduct.”
Autor/es:
• ROLDÁN, M.; JIMÉNEZ DÍAZ, M.; VALDECANTOS, P.; MICELI, D. C.
Lugar:
Villa Carlos Paz, Córdoba, Argentina
Reunión:
Congreso; XXXVII Reunión Anual de la Sociedad Argentina de Investigaciones en Bioquímica y Biología Molecular - SAIB.; 2001
Institución organizadora:
Sociedad Argentina de Investigaciones en Bioquímica y Biología Molecular - SAIB.
Resumen:
PO47. PROTEOLYTIC ACTIVITY AND GENE EXPRESSION OF PLASMINOGEN ACTIVATOR IN PORCINE OVIDUCT Roldán, Mariela; Jiménez Díaz, María; Valdecantos, Pablo and Miceli, Dora. /NS/B/O. UN]: Chacabuco 46/. CP 4000. Tucumán. E-mail: mroldan@mail.unt.edu.ar At present, the physiological role of most oviducal proteins remains unknown. In this work, we present evidence that oviducal secretion as well as oviducal tissue-extract show proteolytic activity. Some of their propertjes were analyzed in Triton X-100 treated tissue crude membrane fraction (Triton-CMF) and in oviducal flushing. We compared the activity of oviducts obtained from immature sows and female in proestrus and diestrus using a chromogenic substrate method. The proteolytic activity was higher in the Triton- CMF oviducts during proestrus than in those of diestrus, while in oviducal flushing, highest activity was observed in diestrus. 100% of the Triton-CMF proteolytic activity was plasminogen-dependent. The use of amiloride, a selective urokinase-type plasminogen activator (uPA) inhibitor, showed that almost 100% of this activity in Triton-CMF was due to a tissue-type plasminogen activator (tPA); in oviducal flushing approximately 60% of the activity was tPA and 40% uPA during diestrus. However SDS-PAGE zymographic analysis in presence of casein and plasminogen reveled a caseinolytic zone at approximately 30 kDa of plasminogen dependent activity that matched with low molecular weight of uPA. Coincidentally, RT-PCR:assays of different sexual cycle stage oviducts using specific pig urokinase primers showed a specific amplified band, confirming that uPA is being synthesized in the pig oviduct. The uPA is probably being seecreted taking into account the activity corresponding to uPA detected in the oviducal flushing.