In membranes of rat olfactory bulb, a brain region in which muscarinic agonists increase cyclic AMP formation, the muscarinic stimulation of guanosine 5'-O-(3-[35S]thiotriphosphate) ([35S]GTP gamma S) binding was used as a tool to investigate the receptor interaction with the guanine nucleotide-binding regulatory proteins (G proteins). The stimulation of the radioligand binding by carbachol (CCh) was optimal (threefold increase) in the presence of micromolar concentrations of GDP and 100 mM NaCl. Exposure to N-ethylmaleimide and pertussis toxin markedly inhibited the CCh effect, whereas it increased the relative stimulation of [35S]GTP gamma S binding elicited by pituitary adenylate cyclase-activating polypeptide (PACAP). On the other hand, membrane treatment with cholera toxin curtailed the PACAP stimulation of [35S]GTP gamma S binding but did not affect the response to CCh. Like CCh, a number of cholinergic agonists stimulated [35S]GTP gamma S binding in a concentration-dependent and saturable manner. The antagonist profile of the muscarinic stimulation of [35S]GTP gamma S binding was highly correlated with that displayed by the muscarinic stimulation of adenylyl cyclase. These data indicate that the olfactory bulb muscarinic receptors couple to G1/G0, but not to Gs, and support the possibility that activation of G1/G0 mediates the stimulatory effect on adenylyl cyclase activity.

STIMULATION OF GUANOSINE-5'-O-(3-[35S]THIO)TRIPHOSPHATE BINDING BY CHOLINERGIC MUSCARINIC RECEPTORS IN MEMBRANES OF RAT OLFACTORY BULB

OLIANAS, MARIA CONCETTA;ONALI, PIER LUIGI
1996-01-01

Abstract

In membranes of rat olfactory bulb, a brain region in which muscarinic agonists increase cyclic AMP formation, the muscarinic stimulation of guanosine 5'-O-(3-[35S]thiotriphosphate) ([35S]GTP gamma S) binding was used as a tool to investigate the receptor interaction with the guanine nucleotide-binding regulatory proteins (G proteins). The stimulation of the radioligand binding by carbachol (CCh) was optimal (threefold increase) in the presence of micromolar concentrations of GDP and 100 mM NaCl. Exposure to N-ethylmaleimide and pertussis toxin markedly inhibited the CCh effect, whereas it increased the relative stimulation of [35S]GTP gamma S binding elicited by pituitary adenylate cyclase-activating polypeptide (PACAP). On the other hand, membrane treatment with cholera toxin curtailed the PACAP stimulation of [35S]GTP gamma S binding but did not affect the response to CCh. Like CCh, a number of cholinergic agonists stimulated [35S]GTP gamma S binding in a concentration-dependent and saturable manner. The antagonist profile of the muscarinic stimulation of [35S]GTP gamma S binding was highly correlated with that displayed by the muscarinic stimulation of adenylyl cyclase. These data indicate that the olfactory bulb muscarinic receptors couple to G1/G0, but not to Gs, and support the possibility that activation of G1/G0 mediates the stimulatory effect on adenylyl cyclase activity.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11584/108153
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