INVESTIGADORES
CAVIEDES VIDAL enrique juan raul
congresos y reuniones científicas
Título:
An improved technique for isolation of brush border vesicles (BBMV) from avian small intestine
Autor/es:
BERDASCO CV; ARIAS R; CAVIEDES VIDAL, E; CHEDIACK, J.G.
Lugar:
San Luis
Reunión:
Congreso; XXX Reunión Anual de la Sociedad de Biología de Cuyo; 2012
Institución organizadora:
Sociedad de Biologia de Cuyo
Resumen:
This study presents an improved method of brush border membrane (BBM) preparation from the avian small intestine using polyethylene glycol (PEG) precipitation. This technique is useful in
physiology because small intestinal brush border membranes contain hydrolytic enzymes and nutrient transporters that are important for the digestion and absorption. Samples were obtained from
adult house sparrows (Passer domesticus) with a body mass around
27 g. The small intestine was excised and divided in three portions
(proximal, medial and distal to the pyloric valve), and the mucosa
was scraped with a glass slide. A 2% homogenate of the mucosa
was prepared in 2 mM Tris?HCl buffer (pH 7.1). A solution of
PEG 3350 was added to obtain a final concentration of 7% and
stirred for 15 min at 4°C. This was centrifuged at 7,500g for 15
min, the supernatant was respun at 12,000g for 15 min and finally
supernatant was respun at 40,000g for 45 min. The pellet was washed
twice with buffer and finally suspended in the same buffer. In the
BBMVs were measured total protein content and tested the activity of marker enzymes, sucrase, maltase (for proximal-medial portion) and N-aminopeptidase (for distal portion). Isolated BBMVs
were relatively pure as judged by an 8 to 20 folds enrichment of
marker enzyme activities.