Pharmacological characterisation of mouse calcitonin and calcitonin receptor‐like receptors reveals differences compared with human receptors

Garelja, Michael L.; Bower, Rebekah L.; Brimble, Margaret A.; Chand, Shanan; Harris, Paul W. R.; Jamaluddin, Muhammad A.; Petersen, Jakeb; Siow, Andrew; Walker, Christopher S.; Hay, Debbie L.

doi:10.1111/bph.15628

Cited by 17 publications

(30 citation statements)

References 57 publications

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“…Antagonist activities of lipidated hαCGRP 8-37 analogues were also tested at mCGRP and AMY 1 receptors as a bridge to in vivo studies in mice. The control agonist in this case, mαCGRP, displayed a potency of 9.59 ± 0.22 (n = 5) at the mCGRP receptor, and 7.82 ± 0.11 (n = 5) at the mAMY 1 receptor, which was slightly lower than the mαCGRP pEC 50 in Garelja et al (2021). The comparatively lower potency of mαCGRP at the mAMY 1 receptor versus the CGRP receptor is different to hαCGRP which is equipotent at both human receptors.…”

Section: Lipidated αCgrp 8-37 Analogues Display Comparable Antagonist...mentioning

confidence: 71%

“…HA-tagged hCLR (Hay et al, 2003) or HA-tagged hCTR (CT (a) splice variant, Udawela et al, 2006) in combination with either myc-tagged hRAMP1 (Qi et al, 2008), FLAG-tagged hRAMP2 (Qi et al, 2013), or untagged hRAMP3 (Hay et al, 2003) were transfected to express the desired calcitonin-family receptor. Untagged mCT (a) , mCLR, mRAMP1, mRAMP2 and mRAMP3 were purchased from Origene and transfected in an identical method to the human receptors into Cos-7 cells (Garelja et al, 2021).…”

Section: Cell Culture and Transfectionmentioning

confidence: 99%

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Lipidated Calcitonin Gene-Related Peptide (CGRP) Peptide Antagonists Retain CGRP Receptor Activity and Attenuate CGRP Action In Vivo

et al. 2022

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Signaling through calcitonin gene-related peptide (CGRP) receptors is associated with pain, migraine, and energy expenditure. Small molecule and monoclonal antibody CGRP receptor antagonists that block endogenous CGRP action are in clinical use as anti-migraine therapies. By comparison, the potential utility of peptide antagonists has received less attention due to suboptimal pharmacokinetic properties. Lipidation is an established strategy to increase peptide half-life in vivo. This study aimed to explore the feasibility of developing lipidated CGRP peptide antagonists that retain receptor antagonist activity in vitro and attenuate endogenous CGRP action in vivo. CGRP peptide analogues based on the archetypal CGRP receptor antagonist, CGRP8-37, were palmitoylated at the N-terminus, position 24, and near the C-terminus at position 35. The antagonist activities of the lipidated peptide analogues were tested in vitro using transfected Cos-7 cells expressing either the human or mouse CGRP receptor, amylin subtype 1 (AMY1) receptor, adrenomedullin (AM) receptors, or calcitonin receptor. Antagonist activities were also evaluated in SK-N-MC cells that endogenously express the human CGRP receptor. Lipidated peptides were then tested for their ability to antagonize endogenous CGRP action in vivo using a capsaicin-induced dermal vasodilation (CIDV) model in C57/BL6J mice. All lipidated peptides except for the C-terminally modified analogue retained potent antagonist activity compared to CGRP8-37 towards the CGRP receptor. The lipidated peptides also retained, and sometimes gained, antagonist activities at AMY1, AM1 and AM2 receptors. Several lipidated peptides produced robust inhibition of CIDV in mice. This study demonstrates that selected lipidated peptide antagonists based on αCGRP8-37 retain potent antagonist activity at the CGRP receptor and are capable of inhibition of endogenous CGRP action in vivo. These findings suggest that lipidation can be applied to peptide antagonists, such as αCGRP8-37 and are a potential strategy for antagonizing CGRP action.

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Section: Lipidated αCgrp 8-37 Analogues Display Comparable Antagonist...mentioning

confidence: 71%

Section: Cell Culture and Transfectionmentioning

confidence: 99%

Lipidated Calcitonin Gene-Related Peptide (CGRP) Peptide Antagonists Retain CGRP Receptor Activity and Attenuate CGRP Action In Vivo

et al. 2022

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“…Thus, it is thought that sCT or its receptor is not involved in the anxiety-related neural circuits. The CTR family is extremely complex, including CTR and CTRlike receptors and three RAMPs (Garelja et al, 2021). AMYR consists of a core CTR and one out of three RAMPs and is expressed in the whole brain including reward-related regions (Kalafateli et al, 2021).…”

Section: Discussionmentioning

confidence: 99%

Salmon Calcitonin Exerts an Antidepressant Effect by Activating Amylin Receptors

Jiang

Ju²,

Luo³

et al. 2022

Front. Pharmacol.

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Depressive disorder is defined as a psychiatric disease characterized by the core symptoms of anhedonia and learned helplessness. Currently, the treatment of depression still calls for medications with high effectiveness, rapid action, and few side effects, although many drugs, including fluoxetine and ketamine, have been approved for clinical usage by the Food and Drug Administration (FDA). In this study, we focused on calcitonin as an amylin receptor polypeptide, of which the antidepressant effect has not been reported, even if calcitonin gene-related peptides have been previously demonstrated to improve depressive-like behaviors in rodents. Here, the antidepressant potential of salmon calcitonin (sCT) was first evaluated in a chronic restraint stress (CRS) mouse model of depression. We observed that the immobility duration in CRS mice was significantly increased during the tail suspension test and forced swimming test. Furthermore, a single administration of sCT was found to successfully rescue depressive-like behaviors in CRS mice. Lastly, AC187 as a potent amylin receptor antagonist was applied to investigate the roles of amylin receptors in depression. We found that AC187 significantly eliminated the antidepressant effects of sCT. Taken together, our data revealed that sCT could ameliorate a depressive-like phenotype probably via the amylin signaling pathway. sCT should be considered as a potential therapeutic candidate for depressive disorder in the future.

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“…Untagged rat CLR, CTR and RAMP1 constructs in pCMV6 were from Origene (Rockville, United States). The untagged mouse CLR, CTR and RAMP1 in pCMV6 plasmids were as previously described ( Garelja et al, 2021a ). Rat and mouse CTR were the CT (a) variant.…”

Section: Methodsmentioning

confidence: 99%

“…Of note, there are species differences in the number of potential CGRP-responsive receptors. More rat and mouse CLR or CTR-based receptors are responsive to CGRP, as compared to their human counterparts ( Hay et al, 2003 ; Bailey et al, 2012 ; Bohn et al, 2017 ; Garelja et al, 2021a ). Therefore, it is important to understand where CTR might be found in relation to CGRP-expressing structures in pre-clinical model species, and in humans.…”

Section: Introductionmentioning

confidence: 99%

CGRP and the Calcitonin Receptor are Co-Expressed in Mouse, Rat and Human Trigeminal Ganglia Neurons

et al. 2022

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The neuropeptide calcitonin gene-related peptide (CGRP) is expressed in the trigeminal ganglia, a key site in craniofacial pain and migraine. CGRP potently activates two receptors: the CGRP receptor and the AMY1 receptor. These receptors are heterodimers consisting of receptor activity-modifying protein 1 (RAMP1) with either the calcitonin receptor-like receptor (CLR) to form the CGRP receptor or the calcitonin receptor (CTR) to form the AMY1 receptor. The expression of the CGRP receptor in trigeminal ganglia has been described in several studies; however, there is comparatively limited data available describing AMY1 receptor expression and in which cellular subtypes it is found. This research aimed to determine the relative distributions of the AMY1 receptor subunit, CTR, and CGRP in neurons or glia in rat, mouse and human trigeminal ganglia. Antibodies against CTR, CGRP and neuronal/glial cell markers were applied to trigeminal ganglia sections to investigate their distribution. CTR-like and CGRP-like immunoreactivity were observed in both discrete and overlapping populations of neurons. In rats and mice, 30–40% of trigeminal ganglia neurons displayed CTR-like immunoreactivity in their cell bodies, with approximately 78–80% of these also containing CGRP-like immunoreactivity. Although human cases were more variable, a similar overall pattern of CTR-like immunoreactivity to rodents was observed in the human trigeminal ganglia. CTR and CGRP appeared to be primarily colocalized in small to medium sized neurons, suggesting that colocalization of CTR and CGRP may occur in C-fiber neurons. CGRP-like or CTR-like immunoreactivity were not typically observed in glial cells. Western blotting confirmed that CTR was expressed in the trigeminal ganglia of all three species. These results confirm that CTR is expressed in trigeminal ganglia neurons. The identification of populations of neurons that express both CGRP and CTR suggests that CGRP could act in an autocrine manner through a CTR-based receptor, such as the AMY1 receptor. Overall, this suggests that a trigeminal ganglia CTR-based receptor may be activated during migraine and could therefore represent a potential target to develop treatments for craniofacial pain and migraine.

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Pharmacological characterisation of mouse calcitonin and calcitonin receptor‐like receptors reveals differences compared with human receptors

Cited by 17 publications

References 57 publications

Lipidated Calcitonin Gene-Related Peptide (CGRP) Peptide Antagonists Retain CGRP Receptor Activity and Attenuate CGRP Action In Vivo

Lipidated Calcitonin Gene-Related Peptide (CGRP) Peptide Antagonists Retain CGRP Receptor Activity and Attenuate CGRP Action In Vivo

Salmon Calcitonin Exerts an Antidepressant Effect by Activating Amylin Receptors

CGRP and the Calcitonin Receptor are Co-Expressed in Mouse, Rat and Human Trigeminal Ganglia Neurons

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