Affiliation:
1. From the Research Center, Centre Hospitalier Universitaire de Montréal-(CHUM), Montreal, Quebec, Canada (S.N.O., J.T., P.H.), and the Laboratory of Biomembranes, Faculty of Biology, Moscow State University, Moscow, Russia (S.N.O., S.L.A., N.I.P.)
Abstract
This study examines the involvement of GTP-binding proteins (Gps) in the regulation of Na
+
/H
+
exchange and Ca
2+
influx, which are increased in vascular smooth muscle cells from spontaneously hypertensive rats. Gp activity was modulated by fluoride, GTPγS, GDPβS, and antisense oligodeoxynucleotides complementary to conserved regions of the α- and β-subunits of Gps (α-comm and β-comm, respectively). β-Adrenergic-induced G
s
-mediated cAMP production was used as a positive control to estimate the efficiency of these compounds. Na
+
/H
+
exchange, measured as ethylisopropyl amiloride-sensitive
22
Na influx, was activated by 5- to 6-fold by a 30-minute preincubation of cells with 10 mmol/L NaF with a
K
0.5
for NaF of _13 mmol/L. In contrast, no activation of
45
Ca influx was observed under preincubation of vascular smooth muscle cells with NaF in Ca
2+
-free medium, whereas at [Ca
2+
]
o
>0.5 mmol/L, simultaneous addition of
45
Ca and 10 mmol/L NaF led to sharply increased isotope uptake. NaF-induced
45
Ca influx did not reach saturation up to 3 mmol/L [Ca
2+
]
0
and 20 mmol/L NaF and was correlated with the formation of calcium-fluoride complexes measured by light scattering. GTPγS increased basal cAMP production and Na
+
/H
+
exchange, whereas GDPβS decreased isoproterenol-induced cAMP production and Na
+
/H
+
exchange, α-comm reduced whereas β-comm augmented isoproterenol-induced cAMP production by 70%. Both oligodeoxynucleotides decreased basal Na
+
/H
+
exchange by 40% to 50%. NaF-induced Na
+
/H
+
exchange was not sensitive to α-comm but was inhibited by 60% in β-comm-loaded cells. Neither basal nor NaF-induced
45
Ca uptake was affected by GTPγS, GDPβS, and the oligodeoxynucleotides. Our results show that
45
Ca uptake is activated by NaF in vascular smooth muscle cells by nonspecific accumulation of calcium-fluoride complexes and is not related to modification of Gps. On the contrary, the Na
+
/H
+
exchanger is controlled by Gps, and Gp β-subunits are involved in [Ca
2+
]
0
-independent activation of this carrier by NaF.
Publisher
Ovid Technologies (Wolters Kluwer Health)
Cited by
2 articles.
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