Characterization of CGRP, and CGRP, Receptor Subtypes" REMI QUIRION," DENISE VAN ROSSUM,b YVAN D U M O N T , ~SERGE ST-PIERRE,' AND ALAIN FOURNIER,' bDepurtriients of Psychiatry, und PhLirmacology und Therapeutics, und Doitglus Hospitd Research Centre McGill University Vordrrn, Quebec, Canadu, H4H 1R3 INRS-SuntP Universiti dm QuPhec Pointe Claire, Qitihec, Canada, H3R 1G6
Calcitonin gene-related peptide (CGRP) is a 37-amino acid peptide generated from the alternate tissue-specific processing of the calcitonin gene mRNA.'.? Various forms (a and p ) of CGRP have been characterized from human tissue^.^-^ In the periphery, CGRP is co-localized and co-released with calcitonin in the parafollicular C cell of the thyroid gland,' with substance P in sensory nerves' and with acetylcholine in motoneurons.* In the brain, CGRP-like immunoreactivity is broadly distributed and especially concentrated in hypothalamic areas and some brainstem nuclei.9 A variety of biological effects have been claimed for CGRP, including the and of modulation of nicotinic receptor activities at the neuromuscularjunction'o*ll substance P in inflammation," a reduction of gastric acid s e c r e t i ~ n , ' ~peripheral .'~ blood vessel dilation,Is cardiac a~celeration,'"'~ a regulation of calcium metabolismh and insulin secretion", an increase in body temperature, and a decrease in food intake. Iy-?' In spite of the various biological actions induced by CGRP, relatively little is Consequently, a few known concerning the characteristics of its receptor~.'~,?'-?~ years ago, our group undertook the development of selective agonists and antagonists which prove to be most useful for the identification of the existence of CGRP receptor s ~ b t y p e s . ' ~ ~ ~ ~ - various " ' T h e criteriaused to classify these receptor classes are reviewed here.
PROFILE OF [C~S(ACM)~~']CW-~CGRP AS A RECEPTOR SUBTYPE SELECTIVE AGONIST Early studies had revealed that the mid-portion of the CGRP molecule, made up of amino acid residues 8 to 18, most likely adopted an amphiphilic alpha helical
I000
> 1000
0.81
11.8
5 .9
> 1000
>I000
0.71 0.82 0.86 4.7
9.1 8.5 8.4
5.8 6.2 6.4 12.1
>loo0 >I000 >loo0
>I000 >loo0 >lo00 > 1000
6.9
8.2 >I000 > lo00 >lo00
9.9
> 1000 > 1000
> 1000 > 1000
> 1000
> 1000
> 1000 > 1000 > 1000
> 1000
>I000
> I000
5.1
I .8
21.1 9.9 5.5
1.9
6.1
26.1 17.2 10.0 9.2
2.2 I .9
5.6 5.6
13.5
2.0 3.2 2.8 2.7 4.2
5.5 5.3 5.2 5.2 8.5
10.7 10.2 11.1 9.7 -
4.I
>1000
> 1000
> 1000 > 1000 > 1000
> 1000
> 1000
NOTE: Data derived from quantitative receptor autoradiography. lCso represents the concentration of competitors required to compete for 50% of specifically bound [''51]a-hCGKP.
to determine whether these putative receptor subtypes are coupled to different transduction mechanisms. We have recently obtained indirect evidence that suggests that CGRP receptors are possibly coupled to different G-proteins in various tissue^.^' This could provide additional evidence for the existence of CGRP receptor subtypes, although recent studies have revealed that a single receptor class can be coupled to multiple transduction pathways. Naturally, the cloning of the respective CGRP receptor subtypes proposed here will provide definite evidence for their existence.
QUIRION e l al.: CGRP RECEPTOR SUBTYPES
103
It would also b e of interest to determine whether the CGRP, and CGRP, receptor subtypes correspond to the previously reported a (I) andp (11)-preferential receptor c l a ~ s e s . * ~Consequently, *~’ w e are currently evaluating the relative affinities and potencies of the a and p forms of both human a n d rat CGRP in a variety it *m a y be of functional assays. Finally, amylin being a CGRP-related ~ e p t i d e , ~ important t o determine whether this newly characterized peptide possesses preferential affinity for a given C G R P receptor class. This information should help t o better characterize t h e functional relevance of CGRP and CGRP receptors in t h e maintenance of body homeostasis.
ACKNOWLEDGMENTS T h e expert secretarial assistance of Mrs. Joan Currie is acknowledged. We thank Drs. A. Cadieux a n d F. Jolicoeur for giving us access t o unpublished results. REFERENCES I.
2.
3.
4. 5. 6.
7.
8.
9. 10.
AMARA,S. G., V. JONAS, M. G. ROSENFELD,E. S . ONG & R. M. EVANS.1982. Alternative RNA processing in calcitonin gene expression generates mRNAs encoding different polypeptide products. Nature 296: 240-244. S . G. AMARA, L. W. SWANSON, P. E. SAWCHENKO, ROSENFELD, M. G., J . J. MERMOD, J . RIvim, W. W. VALE& R. M. EVANS.1983. Production of a novel neuropeptide encoded by the calcitonin gene via tissue-specific RNA processing. Nature 304: 129- 135. MORRIS, H. R., M. PANICO, T. ETIENNE, J. TIPPINS,s. I. GIRGlS & 1. MACINTYRE. 1984. Isolation and characterization of human calcitonin gene-related peptide. Nature 308: 746-748. AMARA,S . G., J . L. ARRIZA,L. W. SWANSON, R. M. EVANS& M. G . ROSENFIELD. 1985. Expression in brain of a messenger RNA encoding a novel neuropeptide homologous to calcitonin gene-related peptide. Science 119: 1094- 1097. STEENBERGH, P. H., J. W. M. HOPPENER, J . ZANDBERG, W. J . VAN DE VEN, H. S . JANSZ& C. J. M. LIPS. 1985. A second calcitoninlCGRP gene. FEBS Lett. 183: 403-407. GRUNDITZ, T., K. EKMAN,K. HAKANSON,C. RERUP,F. SUNDLER & R. UDDMAN. 1986. Calcitonin gene-related peptide in thyroid nerve fibers and C cells: Effects on thyroid hormone secretion and response to hypercalcemia. Endocrinology 119: 23 13-2324. WIESENFELD-HALLEN, Z., T. HOKFELT,J . M. LUNDBERG, W. C. FORSSMANN, M. REINECKE, F. A. TSCHOPP& J . A. FISCHER. 1984. Immunoreactive calcitonin gene-related peptide and substance P coexist in sensory neuron to the spinal cord and interact in spinal behavioral responses of the rat. Neurosci. Lett. 52: 199-204. TAKAMI,K., Y. KAMAL,S. SHIOSAKA, Y . LEE,S . GIRGIS,C. J . HILLYARD, I. MACINTYRE, P. C. EMSON& M. TOHYAMA. 1985. Immunohistochemical evidence for the coexistence of calcitonin gene-related peptide and choline acetyltransferase-like immunoreactivity in neurons of rat hypoglossal, facial and ambiguous nuclei. Brain Res. 328: 385-389. SKOFITSCH, G. & D. M. JACOBowlTZ. 1985. Calcitonin gene-related peptide: Detailed immunohistochemical distribution in the central nervous system. Peptides 6 72 1-745. MILES, K., P. GREENCARD & R. L. HUGANIR. 1989. Calcitonin gene-related peptide
104
II. 12. 13.
14.
15.
16. 17. 18. 19.
20. 21. 22. 23. 24. 25. 26. 27. 28. 29. 30.
ANNALS NEW YORK ACADEMY OF SCIENCES regulates phosphorylation of the nicotinic acetylcholine receptor in rat myotubes. Neuron 2: 1517-1524. NEW, H. V. & A. W. MUDGE.1986. Calcitonin gene-related peptide regulates muscle acetylcholine receptor synthesis. Nature 323: 809-81 1. OHLEN,A , , N. P. WIKLUND, M. G. PERSON & P. HEDQVIST. 1988. Calcitonin generelated peptide desensitizes skeletal muscle arterioles to substance P in uirro. Br. J . Pharmacol. 95: 673-674. HUGHES,J . J., A. S. LEVINE,J. E. MORLEY,B . A . GOSNELL & S. E. SiLvis. 1984. Intraventricular calcitonin gene-related peptide inhibits gastric acid secretion. Peptides 5: 665-668. Y., K . CHIHARA, H. ABE, H. K", T. KITA. Y. KASCHIO & T. F U J I T A . OKIMURA, 1986. Effect of intracerebroventricular administration of rat calcitonin gene-related peptide (CGRP) and [Asn'.']eel calcitonin on gastric acid secretion in the rats. Jpn. J . Pharmacol. 33: 273-277. BRAIN,S. D., T. J . WILLIAM, J. R. TIPPINS, H. R. MORRIS & I. MACINTYRE. 1985. Calcitonin gene-related peptide is a potent vasodilator. Nature 313: 54-56. TSHIKAWA, T., N. OKAMURA, A. SAiTo, T. MASAKI & K. GOTO.1988. Positive inotropic effect of calcitonin gene-related peptide (CGRP) mediated by cyclic AMP in the guinea pig heart. Circ. Res. 63: 726-734. SIGRIST,S . , A. FRANCO-CERECEDA, R. MUFF,H. HENKE,J . M. LUNDBERC & J. A. FISCHER. 1986. Specific receptor and cardiovascular effects of calcitonin gene-related peptide. Endocrinology 119: 381-388. HERMANSEN, K. & B. AHREN.1990. Dual effects of calcitonin gene-related peptide on insulin secretion in the perfused dog pancreas. Reg. Peptides 27: 149-147. DENNIS,T., A. FOURNIER, A . CADIEUX, F. POMERLEAU, F. B. JOLICOEUR, S. ST& R. QUIRION. 1990. hCGRP,,, a calcitonin gene-related peptide antagonist PIERRE revealing CGRP receptor heterogeneity in brain and periphery. J . Pharmucol. Exp. Ther. 254: 123-128. KRAHN,D. D.. B. A. GOSNEI~I., A. S . LEVINE& J. E. MORLEY.1984. Effects of calcitonin gene-related peptide on food intake. Peptides 5 861-864. JOLICOEUR, F. B., D. MENARD, A. FOURNIER, S . ST-PIERRE, A. DRUMHELLER. 1992. Structure activities analysis of CGRPs neurobehavioral effects. This volume. L Y N C HB. , & E. T. KAISER.1988. Biological properties of two models of calcitonin gene-related peptide with idealized amphiphilie a-helices of different lengths. . . - Biochemistry 271 7600-7607. TiPPINS. J . R.. V. M . DIMARZO, M. PANICO.H. R. MORRIS& I . MACINTYRE. 1986. Investigation of the structure-activity relationship of human calcitonin gene-related peptide (CGRP). Biochem. Biophys. Res. Commun. 134: 1306-131 I . SEXTON,P. M., J . S . MCKENZIE & F. A. 0. MENDELSOHN. 1988. Evidence for a new subclass of calcitonin gene-related peptide binding sites in rat brain. Neurochem. Int. 1 2 323-335. UMEDA, Y. & M. ARISAWA. 1989. Characterization of calcitonin gene-related peptide (CGRP) receptors in guinea pig lung. Jpn. J. Pharmacol. 51: 377-384. DOTTI-SIGRIST, S . , W. BORN& J . A. FISCHER. 1988. Identification of a receptor for calcitonin gene-related peptides 1 and I1 in human cerebellum. Biochem. Biophys. Res. Commun. 151: 1081-1087. H E N K ET., , S. SICRIST, W. LANG,J. SCHNEIDER & J. A . FISCHER.1987. Comparison of binding sites for the calcitonin gene-related peptides I and I1 in man. Brain Res. 410: 404-408. DENNIS, T., A. F O U R N I E R , S . ST-PIERRE & R. Q u i R I o N . 1989. Structure-activity profile of calcitonin gene-related peptide in peripheral and brain tissues. Evidence for receptor multiplicity. J . Pharmacol. Exp. Ther. 251: 718-725. DENNIS, T., A. F O U R N I E R , s. GUARD,s. ST-PIERRE & R. QUlRloN. 1991. Cakitonin gene-related peptide (hCGRPa) binding sites in nucleus accumbens. Atypical structural requirements and marked phylogenic differences. Brain Res. 539: 59-66. S. ST-PIERRE & R. Q U I R I O N1991. . ComMIMEAULT, M., A. F O U R N I E R , Y . DUMONT, parative affinities and antagonistic potencies of various human calcitonin gene-related
QUIRION et al.: CGRP RECEPTOR SUBTYPES
31. 32. 33. 34. 35. 36.
37. 38. 39. 40. 41. 42.
105
peptide fragments on CGRP receptors in brain and periphery. J. Pharmacol. Expt. Ther. 258: 1084-1090. SEIFERT,H., J. CHESNUT, E. B. DE SOUZA, J. RIVIER& W. VALE.1985. Binding sites for calcitonin gene-related peptide in distinct areas of rat brain. Brain Res. 346: 195- 198. S. & S. RATTAN.1990. [Tyro]-Calcitoningene-related peptides 28-37 (rat) CHAKDER, as a putative antagonist of calcitonin gene-related peptide responses on opossum internal anal sphincter smooth muscle. J. Pharmacol. Exp. Ther. 253: 200-206. CHIBA,T., A. YAMACUCHI, T. YAMATANI, A. NAKAMURA, T. MORISHITA, T. INUI, 1989. Calcitonin gene-related peptide receptor M. FUKASE,T. NODA& T. FUJITA. antagonist human CGRP. (8-37). Am. J. Physiol. 256 E331-E335. & I. MACINTYRE. INACAKI,S., S. KITO, Y. KUBOTA,S. GIRGIS,C. J. HILLYARD 1986. Autoradiographic localization of calcitonin gene-related peptide binding sites in human and rat brains. Brain Res. 374: 287-298. SEXTON, P. M., J. S. MCKENZIE,R. T. MASON,J. M. MOSELEY,T. MARTIN& F. A. 0. MENDELSOHN. 1986. Localization of binding sites for calcitonin gene-related peptide in rat brain by in uitvo autoradiography. Neuroscience 1 9 1235-1245. TSCHOPP, F. A., H. HENKE,J. B. PETERMANN, P. H. TOBLER, R. JANZER, T. HOKFELT, J. M. LUNDBERG, C. CUELLO & J. A. FISCHER.1985. Calcitonin gene-related peptide and its binding sites in the human central nervous system and pituitary. Proc. Natl. Acad. Sci. USA, 82: 248-252. G. & D. M. JACOBOWITZ. 1985. Autoradiographic distribution of Iz5IcalciSKOFITSCH, tonin gene-related peptide binding sites in the rat central nervous system. Peptides 4 975-986. DUMONT,Y., A. FOURNIER, S. ST-PIERRE, T. W. SCHWARTZ & R. QUIRION.1990. Differential distribution of neuropeptide Y,and Yzreceptors in the rat brain. Eur. J. Pharmacol. 191: 501-503. & B. P. ROQUES.1983. AutoradioQUIRION, R., J. M. ZAJAC,J . L. ZAJAC,J. L. MORGAT graphic distribution of mu and delta opiate receptors in rat brain using highly selective ligands. Life Sci 33(1):227-230. C. NETTI,V. SIBILIA,G. BIELLA& P. C. BRAGA.1987. PECILE,A., F. GUIDOBONO, Calcitonin gene-related peptide: antinociceptive activity in rats, comparison with calcitonin. Regul. Pept. 18: 189-199. VAN ROSSUM,D., Y. DUMONT,A. FOURNIER, S. ST-PIERRE& R. QUIRION.1991. Effect of dithiothreitol and Gpp(NH)p on ['z51]hCGRPabinding sites in brain and peripheral tissues. This volume. M. PARRY-BILLINGS, J. M. COOPER,G. J. S., B. LEIGHTON,G. D. DIMITRIADIS, KOWALCHUK, K. HOWLAND, J. B. ROTHBAND, A. C. WILLIS& K. B. M. REID. 1988. Amylin found in amyloid deposits in human type 2 diabetes mellitus may be a hormone that regulates glycogen metabolism in skeletal muscle. Proc. Natl. Acad. Sci. USA 85: 7763-7766.
Calcitonin gene-related peptide (CGRP) is a 37-amino acid peptide generated from the alternate tissue-specific processing of the calcitonin gene mRNA.'.? Various forms (a and p ) of CGRP have been characterized from human tissue^.^-^ In the periphery, CGRP is co-localized and co-released with calcitonin in the parafollicular C cell of the thyroid gland,' with substance P in sensory nerves' and with acetylcholine in motoneurons.* In the brain, CGRP-like immunoreactivity is broadly distributed and especially concentrated in hypothalamic areas and some brainstem nuclei.9 A variety of biological effects have been claimed for CGRP, including the and of modulation of nicotinic receptor activities at the neuromuscularjunction'o*ll substance P in inflammation," a reduction of gastric acid s e c r e t i ~ n , ' ~peripheral .'~ blood vessel dilation,Is cardiac a~celeration,'"'~ a regulation of calcium metabolismh and insulin secretion", an increase in body temperature, and a decrease in food intake. Iy-?' In spite of the various biological actions induced by CGRP, relatively little is Consequently, a few known concerning the characteristics of its receptor~.'~,?'-?~ years ago, our group undertook the development of selective agonists and antagonists which prove to be most useful for the identification of the existence of CGRP receptor s ~ b t y p e s . ' ~ ~ ~ ~ - various " ' T h e criteriaused to classify these receptor classes are reviewed here.
PROFILE OF [C~S(ACM)~~']CW-~CGRP AS A RECEPTOR SUBTYPE SELECTIVE AGONIST Early studies had revealed that the mid-portion of the CGRP molecule, made up of amino acid residues 8 to 18, most likely adopted an amphiphilic alpha helical
I000
> 1000
0.81
11.8
5 .9
> 1000
>I000
0.71 0.82 0.86 4.7
9.1 8.5 8.4
5.8 6.2 6.4 12.1
>loo0 >I000 >loo0
>I000 >loo0 >lo00 > 1000
6.9
8.2 >I000 > lo00 >lo00
9.9
> 1000 > 1000
> 1000 > 1000
> 1000
> 1000
> 1000 > 1000 > 1000
> 1000
>I000
> I000
5.1
I .8
21.1 9.9 5.5
1.9
6.1
26.1 17.2 10.0 9.2
2.2 I .9
5.6 5.6
13.5
2.0 3.2 2.8 2.7 4.2
5.5 5.3 5.2 5.2 8.5
10.7 10.2 11.1 9.7 -
4.I
>1000
> 1000
> 1000 > 1000 > 1000
> 1000
> 1000
NOTE: Data derived from quantitative receptor autoradiography. lCso represents the concentration of competitors required to compete for 50% of specifically bound [''51]a-hCGKP.
to determine whether these putative receptor subtypes are coupled to different transduction mechanisms. We have recently obtained indirect evidence that suggests that CGRP receptors are possibly coupled to different G-proteins in various tissue^.^' This could provide additional evidence for the existence of CGRP receptor subtypes, although recent studies have revealed that a single receptor class can be coupled to multiple transduction pathways. Naturally, the cloning of the respective CGRP receptor subtypes proposed here will provide definite evidence for their existence.
QUIRION e l al.: CGRP RECEPTOR SUBTYPES
103
It would also b e of interest to determine whether the CGRP, and CGRP, receptor subtypes correspond to the previously reported a (I) andp (11)-preferential receptor c l a ~ s e s . * ~Consequently, *~’ w e are currently evaluating the relative affinities and potencies of the a and p forms of both human a n d rat CGRP in a variety it *m a y be of functional assays. Finally, amylin being a CGRP-related ~ e p t i d e , ~ important t o determine whether this newly characterized peptide possesses preferential affinity for a given C G R P receptor class. This information should help t o better characterize t h e functional relevance of CGRP and CGRP receptors in t h e maintenance of body homeostasis.
ACKNOWLEDGMENTS T h e expert secretarial assistance of Mrs. Joan Currie is acknowledged. We thank Drs. A. Cadieux a n d F. Jolicoeur for giving us access t o unpublished results. REFERENCES I.
2.
3.
4. 5. 6.
7.
8.
9. 10.
AMARA,S. G., V. JONAS, M. G. ROSENFELD,E. S . ONG & R. M. EVANS.1982. Alternative RNA processing in calcitonin gene expression generates mRNAs encoding different polypeptide products. Nature 296: 240-244. S . G. AMARA, L. W. SWANSON, P. E. SAWCHENKO, ROSENFELD, M. G., J . J. MERMOD, J . RIvim, W. W. VALE& R. M. EVANS.1983. Production of a novel neuropeptide encoded by the calcitonin gene via tissue-specific RNA processing. Nature 304: 129- 135. MORRIS, H. R., M. PANICO, T. ETIENNE, J. TIPPINS,s. I. GIRGlS & 1. MACINTYRE. 1984. Isolation and characterization of human calcitonin gene-related peptide. Nature 308: 746-748. AMARA,S . G., J . L. ARRIZA,L. W. SWANSON, R. M. EVANS& M. G . ROSENFIELD. 1985. Expression in brain of a messenger RNA encoding a novel neuropeptide homologous to calcitonin gene-related peptide. Science 119: 1094- 1097. STEENBERGH, P. H., J. W. M. HOPPENER, J . ZANDBERG, W. J . VAN DE VEN, H. S . JANSZ& C. J. M. LIPS. 1985. A second calcitoninlCGRP gene. FEBS Lett. 183: 403-407. GRUNDITZ, T., K. EKMAN,K. HAKANSON,C. RERUP,F. SUNDLER & R. UDDMAN. 1986. Calcitonin gene-related peptide in thyroid nerve fibers and C cells: Effects on thyroid hormone secretion and response to hypercalcemia. Endocrinology 119: 23 13-2324. WIESENFELD-HALLEN, Z., T. HOKFELT,J . M. LUNDBERG, W. C. FORSSMANN, M. REINECKE, F. A. TSCHOPP& J . A. FISCHER. 1984. Immunoreactive calcitonin gene-related peptide and substance P coexist in sensory neuron to the spinal cord and interact in spinal behavioral responses of the rat. Neurosci. Lett. 52: 199-204. TAKAMI,K., Y. KAMAL,S. SHIOSAKA, Y . LEE,S . GIRGIS,C. J . HILLYARD, I. MACINTYRE, P. C. EMSON& M. TOHYAMA. 1985. Immunohistochemical evidence for the coexistence of calcitonin gene-related peptide and choline acetyltransferase-like immunoreactivity in neurons of rat hypoglossal, facial and ambiguous nuclei. Brain Res. 328: 385-389. SKOFITSCH, G. & D. M. JACOBowlTZ. 1985. Calcitonin gene-related peptide: Detailed immunohistochemical distribution in the central nervous system. Peptides 6 72 1-745. MILES, K., P. GREENCARD & R. L. HUGANIR. 1989. Calcitonin gene-related peptide
104
II. 12. 13.
14.
15.
16. 17. 18. 19.
20. 21. 22. 23. 24. 25. 26. 27. 28. 29. 30.
ANNALS NEW YORK ACADEMY OF SCIENCES regulates phosphorylation of the nicotinic acetylcholine receptor in rat myotubes. Neuron 2: 1517-1524. NEW, H. V. & A. W. MUDGE.1986. Calcitonin gene-related peptide regulates muscle acetylcholine receptor synthesis. Nature 323: 809-81 1. OHLEN,A , , N. P. WIKLUND, M. G. PERSON & P. HEDQVIST. 1988. Calcitonin generelated peptide desensitizes skeletal muscle arterioles to substance P in uirro. Br. J . Pharmacol. 95: 673-674. HUGHES,J . J., A. S. LEVINE,J. E. MORLEY,B . A . GOSNELL & S. E. SiLvis. 1984. Intraventricular calcitonin gene-related peptide inhibits gastric acid secretion. Peptides 5: 665-668. Y., K . CHIHARA, H. ABE, H. K", T. KITA. Y. KASCHIO & T. F U J I T A . OKIMURA, 1986. Effect of intracerebroventricular administration of rat calcitonin gene-related peptide (CGRP) and [Asn'.']eel calcitonin on gastric acid secretion in the rats. Jpn. J . Pharmacol. 33: 273-277. BRAIN,S. D., T. J . WILLIAM, J. R. TIPPINS, H. R. MORRIS & I. MACINTYRE. 1985. Calcitonin gene-related peptide is a potent vasodilator. Nature 313: 54-56. TSHIKAWA, T., N. OKAMURA, A. SAiTo, T. MASAKI & K. GOTO.1988. Positive inotropic effect of calcitonin gene-related peptide (CGRP) mediated by cyclic AMP in the guinea pig heart. Circ. Res. 63: 726-734. SIGRIST,S . , A. FRANCO-CERECEDA, R. MUFF,H. HENKE,J . M. LUNDBERC & J. A. FISCHER. 1986. Specific receptor and cardiovascular effects of calcitonin gene-related peptide. Endocrinology 119: 381-388. HERMANSEN, K. & B. AHREN.1990. Dual effects of calcitonin gene-related peptide on insulin secretion in the perfused dog pancreas. Reg. Peptides 27: 149-147. DENNIS,T., A. FOURNIER, A . CADIEUX, F. POMERLEAU, F. B. JOLICOEUR, S. ST& R. QUIRION. 1990. hCGRP,,, a calcitonin gene-related peptide antagonist PIERRE revealing CGRP receptor heterogeneity in brain and periphery. J . Pharmucol. Exp. Ther. 254: 123-128. KRAHN,D. D.. B. A. GOSNEI~I., A. S . LEVINE& J. E. MORLEY.1984. Effects of calcitonin gene-related peptide on food intake. Peptides 5 861-864. JOLICOEUR, F. B., D. MENARD, A. FOURNIER, S . ST-PIERRE, A. DRUMHELLER. 1992. Structure activities analysis of CGRPs neurobehavioral effects. This volume. L Y N C HB. , & E. T. KAISER.1988. Biological properties of two models of calcitonin gene-related peptide with idealized amphiphilie a-helices of different lengths. . . - Biochemistry 271 7600-7607. TiPPINS. J . R.. V. M . DIMARZO, M. PANICO.H. R. MORRIS& I . MACINTYRE. 1986. Investigation of the structure-activity relationship of human calcitonin gene-related peptide (CGRP). Biochem. Biophys. Res. Commun. 134: 1306-131 I . SEXTON,P. M., J . S . MCKENZIE & F. A. 0. MENDELSOHN. 1988. Evidence for a new subclass of calcitonin gene-related peptide binding sites in rat brain. Neurochem. Int. 1 2 323-335. UMEDA, Y. & M. ARISAWA. 1989. Characterization of calcitonin gene-related peptide (CGRP) receptors in guinea pig lung. Jpn. J. Pharmacol. 51: 377-384. DOTTI-SIGRIST, S . , W. BORN& J . A. FISCHER. 1988. Identification of a receptor for calcitonin gene-related peptides 1 and I1 in human cerebellum. Biochem. Biophys. Res. Commun. 151: 1081-1087. H E N K ET., , S. SICRIST, W. LANG,J. SCHNEIDER & J. A . FISCHER.1987. Comparison of binding sites for the calcitonin gene-related peptides I and I1 in man. Brain Res. 410: 404-408. DENNIS, T., A. F O U R N I E R , S . ST-PIERRE & R. Q u i R I o N . 1989. Structure-activity profile of calcitonin gene-related peptide in peripheral and brain tissues. Evidence for receptor multiplicity. J . Pharmacol. Exp. Ther. 251: 718-725. DENNIS, T., A. F O U R N I E R , s. GUARD,s. ST-PIERRE & R. QUlRloN. 1991. Cakitonin gene-related peptide (hCGRPa) binding sites in nucleus accumbens. Atypical structural requirements and marked phylogenic differences. Brain Res. 539: 59-66. S. ST-PIERRE & R. Q U I R I O N1991. . ComMIMEAULT, M., A. F O U R N I E R , Y . DUMONT, parative affinities and antagonistic potencies of various human calcitonin gene-related
QUIRION et al.: CGRP RECEPTOR SUBTYPES
31. 32. 33. 34. 35. 36.
37. 38. 39. 40. 41. 42.
105
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