Inhibition of adenylyl cyclase activity is one of at least four mechanisms by which the neuropeptide galanin inhibits insulin secretion from pancreatic β-cells. In a membrane preparation of the insulin-secreting cell line RINm5F, a maximally effective concentration of galanin inhibited forskolin-stimulated adenylyl cyclase activity by 30%. Pretreatment of the cells with pertussis toxin abolished the inhibitory effect of galanin, indicating the involvement of Gi or Go guanine nucleotide binding proteins (G-proteins). Because galanin receptors interact with four G-proteins (Gi1, Gi2, Gi3, and Go1), any or all of these may inhibit adenylyl cyclase. Therefore, to identify the G-protein(s) involved, antibodies raised against various G-protein alpha-subunits were used to block the inhibition of forskolin-stimulated adenylyl cyclase activity by galanin in RINm5F membrane preparations. Antisera AS/7 and EC/2, specific for Gɑi1/ɑi2 and Gɑi3, respectively, were able to significantly attenuate the inhibitory effect of galanin, whereas antisera specific for Go proteins were not. The use of additional antisera specific for the various subtypes of Gi proteins indicated that Gi2 and Gi3, but not Gi1, are involved. Simultaneous application of antisera AS/7 and EC/2 resulted in a greater attenuation of the effect of galanin than application of either antiserum alone. Thus, galanin inhibition of adenylyl cyclase activity in these cells is selectively mediated by two inhibitory G-proteins, Gi2 and Gi3.
Skip Nav Destination
Article navigation
Original Articles|
April 01 1995
Gi2 and Gi3 Proteins Mediate the Inhibition of Adenylyl Cyclase by Galanin in the RINm5F Cell Free
Alison M McDermott;
Alison M McDermott
Department of Pharmacology, College of Veterinary Medicine, Cornell University
Ithaca, New York
Search for other works by this author on:
Geoffrey W G Sharp
Geoffrey W G Sharp
Department of Pharmacology, College of Veterinary Medicine, Cornell University
Ithaca, New York
Search for other works by this author on:
Diabetes 1995;44(4):453–459
Article history
Received:
April 14 1994
Revision Received:
December 22 1994
Accepted:
December 22 1994
PubMed:
7535266
Citation
Alison M McDermott, Geoffrey W G Sharp; Gi2 and Gi3 Proteins Mediate the Inhibition of Adenylyl Cyclase by Galanin in the RINm5F Cell. Diabetes 1 April 1995; 44 (4): 453–459. https://doi.org/10.2337/diab.44.4.453
Download citation file:
61
Views