INVESTIGADORES
MILESI Maria Veronica
congresos y reuniones científicas
Título:
The human umbilical artery presents a capacitative response induced by intracellular Ca2+ stores depletion and expresses transient receptor protein channels type 3 and 6
Autor/es:
ENRIQUE N; SALEMME S; SPERONI F; REBOLLEDO A; RIMORINI L; ROLDÁN PALOMO R; PETRUCELLI S; DRUT R; MILESIV
Lugar:
Buenos Aires
Reunión:
Congreso; XXII Congreso Latinoamericano y I Ibero-americano de Ciencias Fisiológicas; 2006
Resumen:
In certain cell types,
the depletion of intracellular Ca2+ stores triggers extracellular Ca2+ influx
through non-selective cationic channels (NSCC) allowing the refilling of the
stores, a phenomenon known as capacitative Ca2+ entry. The aim of this work was
to explore the presence of this response in the human umbilical artery (HUA).
Intracellular Ca2+ measurement in HUA segments using fura-4AM indicate that treatment
with 1 µM thapsigargin, which produces sarcoplasmic reticulum depletion,
induced a Ca2+ influx (∆ fluorescence ratio = 0.09 ± 0.01, p<0.05, n=5)
which was blunted by 200 µM Gd3+, a NSCC blocker (n=4). Using the
single-channel patch-clamp technique in the cell-attached configuration, with a
pipette and bath solution containing 140 mM K+, we found that in HUA smooth muscle
cells 5 µM thapsigargin increased the open probability of an ionic channel
which carried an inward current measuring -0.41 ± 0.16 pA at a membrane
potential of -40 mV (n=13). In cells not treated with thapsigargin the activity
of this channel did not increase during a 20 min period (n=8). Similar results were
obtained with cyclopiazonic acid, which also depletes the sarcoplasmic
reticulum (n=9). Transient receptor protein channels (TRPC) are proposed in the
literature to be involved in the capacitative response. Hence we studied if
they were present in the HUA smooth muscle cells. Using specific antibodies, we
demonstrated with immunohistochemical and western blot techniques the presence
of TRPC types 3 and 6 and the absence of type 4. We conclude that this artery
presents a capacitative response and expresses at least two types of channels
which have been implicated in this mechanism