Cloning, characterization and central nervous system distribution of receptor activity modifying proteins in the rat

Oliver, Kevin R.; Kane, Stefanie A.; Salvatore, Christopher; Mallee, John; Kinsey, Anna; Koblan, Kenneth S.; Keyvan-Fouladi, N.; Heavens, R.P.; Wainwright, Anna; Jacobson, Marlene A.; Dickerson, Ian M.; Hill, R.G.

doi:10.1046/j.0953-816x.2001.01688.x

Cited by 102 publications

(88 citation statements)

References 30 publications

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“…Neither 10 nM h␣CGRP nor hAM elicited significant increases in cAMP in nontransfected HEK-293 cells or cells transfected with HA-hRAMP1 or hCRLR alone. On the other hand, the coexpression of hCRLR with HA-hRAMP1 enabled CGRP and AM to each elicit significant increases in cAMP, which is consistent with previous reports that CGRP and AM bind to the CRLR/RAMP1 heterodimer with similar affinities (8,9,12,24,25). Similar CGRPand AM-evoked increases in cAMP levels were seen when hCRLR was co-expressed with D74-76, D105-107, or D109-112.…”

Section: Construction and Characterization Of Hramp1 Deletion Mutants-supporting

confidence: 78%

Identification of the Human Receptor Activity-modifying Protein 1 Domains Responsible for Agonist Binding Specificity

Kuwasako

Kïtamura

Nagoshi

et al. 2003

Journal of Biological Chemistry

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When co-expressed with receptor activity-modifying protein (RAMP) 1, calcitonin receptor-like receptor (CRLR) can function as a receptor for both calcitonin gene-related peptide (CGRP) and adrenomedullin (AM). To investigate the structural determinants of ligand binding specificity, we examined the extracellular domain of human (h) RAMP1 using various deletion mutants. Co-expression of the hRAMP1 mutants with hCRLR in HEK-293 cells revealed that deletion of residues 91-94, 96 -100, or 101-103 blocked [ 125 I]CGRP binding and completely abolished intracellular cAMP accumulation normally elicited by CGRP or AM. On the other hand, the deletion of residues 78 -80 or 88 -90 significantly attenuated only AM-evoked responses. In all of these cases, the receptor heterodimers were fully expressed at the cell surface. Substituting alanine for residues 91-103 one at a time had little effect on CGRPinduced responses, indicating that although this segment is essential for high affinity agonist binding to the receptors, none of the residues directly interacts with either CGRP or AM. This finding suggests that RAMPs probably determine ligand specificity by contributing to the structure of the ligand-binding pocket or by allosteric modulation of the conformation of the receptor. Interestingly, the L94A mutant up-regulated surface expression of the receptor heterodimer to a greater degree than wild-type hRAMP1, thereby increasing CGRP binding and signaling. L94A also significantly increased cell surface expression of the hRAMP1 deletion mutant D101-103 when co-transfected with hCRLR, and expression of a L94A/D101-103 double mutant markedly attenuated the activity of endogenous RAMP1 in HEK-293T cells.CGRP 1 and AM belong to the calcitonin family of regulatory peptides and are both highly potent vasodilators (1, 2). Although they share very little sequence identity, both contain ring structures comprised of six amino acids linked by a disulfide bridge and an amidated C terminus that are required for biological activity (3). Both of these peptides and their specific or common receptors are widely distributed among peripheral tissues and in the central nervous system, enabling them to exert a wide variety of biological effects (4, 5).RAMPs are a recently identified group of single transmembrane domain accessory proteins that serve to transport CRLR to the cell surface where they form functional CGRP and AM receptors (6). The three RAMP isoforms (RAMP1, RAMP2, and RAMP3), which share only 30% sequence identity and differ in their tissue distributions, are all comprised of ϳ160 amino acids that make up a large extracellular N-terminal domain, a single membrane-spanning domain, and a very short cytoplasmic domain (6, 7). Co-expression of CRLR with RAMP1 leads to both proteins being presented at the plasma membrane as a heterodimeric CGRP receptor, whereas co-expression of CRLR with RAMP2 or RAMP3 enables the resultant heterodimer to function as a AM receptor (6,8,9). Upon binding their respective agonist, both receptors mediate a rise...

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Section: Construction and Characterization Of Hramp1 Deletion Mutants-supporting

confidence: 78%

Identification of the Human Receptor Activity-modifying Protein 1 Domains Responsible for Agonist Binding Specificity

Kuwasako

Kïtamura

Nagoshi

et al. 2003

Journal of Biological Chemistry

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“…CLR and RAMP1 were co-expressed, suggesting expression of functional CGRP receptor in fibers within laminae I and II. The presence of receptor components in the spinal cord is supported by the mRNA expression of RAMP1 and RCP, detected with specific oligonucleotides for in situ hybridization [35]. …”

Section: Discussionmentioning

confidence: 99%

Calcitonin gene-related peptide (CGRP) and its receptor components in human and rat spinal trigeminal nucleus and spinal cord at C1-level

2011

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BackgroundCalcitonin gene-related peptide (CGRP) has a key role in migraine pathophysiology and is associated with activation of the trigeminovascular system. The trigeminal ganglion, storing CGRP and its receptor components, projects peripheral to the intracranial vasculature and central to regions in the brainstem with Aδ- and C-fibers; this constitutes an essential part of the pain pathways activated in migraine attacks. Therefore it is of importance to identify the regions within the brainstem that processes nociceptive information from the trigeminovascular system, such as the spinal trigeminal nucleus (STN) and the C1-level of the spinal cord. Immunohistochemistry was used to study the distribution and relation between CGRP and its receptor components - calcitonin receptor-like receptor (CLR) and receptor activity modifying protein 1 (RAMP1) - in human and rat STN and at the C1-level, using a set of newly well characterized antibodies. In addition, double-stainings with CGRP and myelin basic protein (MBP, myelin), synaptophysin (synaptic vesicles) or IB4 (C-fibers in general) were performed.ResultsIn the STN, the highest density of CGRP immunoreactive fibers were found in a network around fiber bundles in the superficial laminae. CLR and RAMP1 expression were predominately found in fibers in the spinal trigeminal tract region, with some fibers spanning into the superficial laminae. Co-localization between CGRP and its receptor components was not noted. In C1, CGRP was expressed in fibers of laminae I and II. The CGRP staining was similar in rat, except for CGRP positive neurons that were found close to the central canal. In C1, the receptor components were detected in laminae I and II, however these fibers were distinct from fibers expressing CGRP as verified by confocal microscopy.ConclusionsThis study demonstrates the detailed expression of CGRP and its receptor components within STN in the brainstem and in the spinal cord at C1-level, and shows the possibility of CGRP acting postjunctionally in these areas putatively involved in primary headaches.

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“…The current qPCR results provide quantifiable data that support this idea, as mRNA for all components of the amylin receptor complex are expressed within the VTA, including both subtypes of CTR and all three RAMP subtypes. Although further investigation is needed to identify the phenotype(s) of VTA neurons (eg, dopaminergic, GABAergic, glutamatergic) that express the amylin receptor complex, the equivalent expression of the three RAMP subtypes within the VTA contrasts with the prevalence of RAMP1 mRNA expression in the NAc (Oliver et al, 2001), as well as with the prevalence of RAMP2 and RAMP3 in the AP (Barth et al, 2004). Different combinations of RAMP and CTR subtypes create receptor complexes that all bind amylin but also have differing affinities for other calcitonin family ligands such as calcitonin generelated peptide and adrenomedullin.…”

Section: Discussionmentioning

confidence: 99%

“…As a preliminary step toward testing a role for VTA amylin receptor signaling in the control of food intake, we used qPCR to identify mRNA expression of the components of the amylin receptor complex in the VTA. In addition, we examined mRNA expression of the two CTR subtypes and the three RAMP subtypes in the NAc core and shell subregions as positive controls, as the NAc expresses amylin receptor complexes consisting primarily of CTR-A and RAMP1 (Oliver et al, 2001;Young, 2005). qPCR analysis revealed that both CTR subtypes and all three RAMP subtypes are expressed in each of the three sites examined, albeit at varying levels of expression.…”

Section: All Components Of the Amylin Receptor Complex Are Expressed mentioning

confidence: 99%

Amylin Receptor Signaling in the Ventral Tegmental Area is Physiologically Relevant for the Control of Food Intake

Mietlicki‐Baase

Rupprecht

Olivos

et al. 2013

Neuropsychopharmacol

116

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The ability of amylin, a pancreatic b-cell-derived neuropeptide, to promote negative energy balance has been ascribed to neural activation at the area postrema. However, despite amylin binding throughout the brain, the possible role of amylin signaling at other nuclei in the control of food intake has been largely neglected. We show that mRNA for all components of the amylin receptor complex is expressed in the ventral tegmental area (VTA), a mesolimbic structure mediating food intake and reward. Direct activation of VTA amylin receptors reduces the intake of chow and palatable sucrose solution in rats. This effect is mediated by reductions in meal size and is not due to nausea/malaise or prolonged suppression of locomotor activity. VTA amylin receptor activation also reduces sucrose selfadministration on a progressive ratio schedule. Finally, antagonist studies provide novel evidence that VTA amylin receptor blockade increases food intake and attenuates the intake-suppressive effects of a peripherally administered amylin analog, suggesting that amylin receptor signaling in the VTA is physiologically relevant for food intake control and potentially clinically relevant for the treatment of obesity.

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Cloning, characterization and central nervous system distribution of receptor activity modifying proteins in the rat

Cited by 102 publications

References 30 publications

Identification of the Human Receptor Activity-modifying Protein 1 Domains Responsible for Agonist Binding Specificity

Identification of the Human Receptor Activity-modifying Protein 1 Domains Responsible for Agonist Binding Specificity

Calcitonin gene-related peptide (CGRP) and its receptor components in human and rat spinal trigeminal nucleus and spinal cord at C1-level

Amylin Receptor Signaling in the Ventral Tegmental Area is Physiologically Relevant for the Control of Food Intake

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