A comparison of two alternatively spliced forms of a metabotropic glutamate receptor coupled to phosphoinositide turnover.
A comparison of the pharmacological and physiological properties of the metabotropic glutamate 1 alpha and 1 beta receptors (mGluR1 alpha and mGluR1 beta) expressed in baby hamster kidney (BHK 570) cells was performed. The mGluR1 beta receptor is an alternatively spliced form of mGluR1 alpha with a modified carboxy terminus. Immunoblots of membranes from the two cell lines probed with receptor-specific antipeptide antibodies showed that mGluR1 alpha migrated with an M(r) = 154,000, whereas mGluR1 beta migrated with an M(r) = 96,000. Immunofluorescence imaging of receptors expressed in BHK 570 cells revealed that the mGluR1 alpha receptor was localized to patches along the plasmalemma and on intracellular membranes surrounding the nucleus, whereas mGluR1 beta was distributed diffusely throughout the cell. Agonist activation of the mGluR1 alpha and the mGluR1 beta receptors stimulated phosphoinositide hydrolysis. At both receptors, glutamate, quisqualate, and ibotenate were full agonists, whereas trans-(+)-1-aminocyclopentane-1,3-dicarboxylate appeared to act as a partial agonist. The stimulation of phosphoinositide hydrolysis by mGluR1 alpha showed pertussis toxin-sensitive and insensitive components, whereas the mGluR1 beta response displayed only the toxin-insensitive component. The mGluR1 alpha and mGluR1 beta receptors also increased intracellular calcium levels by inducing release from intracellular stores. These results indicate that the different carboxy terminal sequences of the two receptors directly influences G protein coupling and subcellular deposition of the receptor polypeptides and suggest that the two receptors may subserve different roles in the nervous system.
Faculty of Pharmacy, University of Toronto, Ontario, Canada.
D S
DS
Pickering
C
C
Thomsen
P D
PD
Suzdak
E J
EJ
Fletcher
R
R
Robitaille
M W
MW
Salter
J F
JF
MacDonald
X P
XP
Huang
D R
DR
Hampson
D000818:Animals; D000918:Antibody Specificity; D002118:Calcium / Q000378:metabolism; D002461:Cell Line, Transformed; D004274:DNA, Recombinant; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D006868:Hydrolysis; D015151:Immunoblotting; D007425:Intracellular Membranes / Q000378:metabolism; D007668:Kidney / Q000166:cytology / Q000378:metabolism; D009994:Osmolar Concentration; D037342:Pertussis Toxin; D010716:Phosphatidylinositols / Q000378:metabolism*; D017470:Receptors, Glutamate / Q000276:immunology / Q000378:metabolism*; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
D004274:DNA, Recombinant; D010716:Phosphatidylinositols; D017470:Receptors, Glutamate; D010566:Virulence Factors, Bordetella; D037342:Pertussis Toxin; D002118:Calcium