Tissue-Specific Expression of GLP1R in Mice: Is the Problem of Antibody Nonspecificity Solved?

Aroor, Annayya R.; Nistala, Ravi

doi:10.2337/db13-1937

Cited by 11 publications

(10 citation statements)

References 18 publications

(16 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Antagonist-linked fluorophores circumvent this issue, but the majority lack thorough pharmacological validation, or possess near infrared tags which require sophisticated confocal imaging modalities. On the other hand, reporter mouse strategies possess high fidelity, but cannot account for lineage-tracing artefacts, post-translational processing, protein stability and trafficking of native receptor 28 . Lastly, none of the aforementioned approaches are amenable to super-resolution imaging of endogenous GLP1R.…”

mentioning

confidence: 99%

Super-resolution microscopy compatible fluorescent probes reveal endogenous glucagon-like peptide-1 receptor distribution and dynamics

et al. 2020

View full text Add to dashboard Cite

The glucagon-like peptide-1 receptor (GLP1R) is a class B G protein-coupled receptor (GPCR) involved in metabolism. Presently, its visualization is limited to genetic manipulation, antibody detection or the use of probes that stimulate receptor activation. Herein, we present LUXendin645, a far-red fluorescent GLP1R antagonistic peptide label. LUXendin645 produces intense and specific membrane labeling throughout live and fixed tissue. GLP1R signaling can additionally be evoked when the receptor is allosterically modulated in the presence of LUXendin645. Using LUXendin645 and LUXendin651, we describe islet, brain and hESC-derived β-like cell GLP1R expression patterns, reveal higher-order GLP1R organization including membrane nanodomains, and track single receptor subpopulations. We furthermore show that the LUXendin backbone can be optimized for intravital two-photon imaging by installing a red fluorophore. Thus, our super-resolution compatible labeling probes allow visualization of endogenous GLP1R, and provide insight into class B GPCR distribution and dynamics both in vitro and in vivo.

show abstract

mentioning

confidence: 99%

Super-resolution microscopy compatible fluorescent probes reveal endogenous glucagon-like peptide-1 receptor distribution and dynamics

et al. 2020

View full text Add to dashboard Cite

show abstract

“…Clear proof of whether this is indeed the case would be successful labelling with a GLP-1R antibody. However, the paucity of reliable GLP-1R antibodies is well established [21,22] , and, consequently, a functional approach was taken in this study. We screened tdRFP-positive cells in a few brain regions with established (e.g.…”

Section: Discussionmentioning

confidence: 99%

Distribution and characterisation of Glucagon-like peptide-1 receptor expressing cells in the mouse brain

Cork

Richards

Holt

et al. 2015

Molecular Metabolism

347

326

View full text Add to dashboard Cite

ObjectiveAlthough Glucagon-like peptide 1 is a key regulator of energy metabolism and food intake, the precise location of GLP-1 receptors and the physiological relevance of certain populations is debatable. This study investigated the novel GLP-1R-Cre mouse as a functional tool to address this question.MethodsMice expressing Cre-recombinase under the Glp1r promoter were crossed with either a ROSA26 eYFP or tdRFP reporter strain to identify GLP-1R expressing cells. Patch-clamp recordings were performed on tdRFP-positive neurons in acute coronal brain slices from adult mice and selective targeting of GLP-1R cells in vivo was achieved using viral gene delivery.ResultsLarge numbers of eYFP or tdRFP immunoreactive cells were found in the circumventricular organs, amygdala, hypothalamic nuclei and the ventrolateral medulla. Smaller numbers were observed in the nucleus of the solitary tract and the thalamic paraventricular nucleus. However, tdRFP positive neurons were also found in areas without preproglucagon-neuronal projections like hippocampus and cortex. GLP-1R cells were not immunoreactive for GFAP or parvalbumin although some were catecholaminergic. GLP-1R expression was confirmed in whole-cell recordings from BNST, hippocampus and PVN, where 100 nM GLP-1 elicited a reversible inward current or depolarisation. Additionally, a unilateral stereotaxic injection of a cre-dependent AAV into the PVN demonstrated that tdRFP-positive cells express cre-recombinase facilitating virally-mediated eYFP expression.ConclusionsThis study is a comprehensive description and phenotypic analysis of GLP-1R expression in the mouse CNS. We demonstrate the power of combining the GLP-1R-CRE mouse with a virus to generate a selective molecular handle enabling future in vivo investigation as to their physiological importance.

show abstract

“…However, earlier studies used RT‐PCR, immunohistochemistry and Western blotting to detect GLP‐1 expression in the heart (Ban et al . ), and the specificity and accuracy of commercially available GLP‐1R antibodies have been questioned (Aroor and Nistala, ). Nevertheless, GLP‐1 administration increased glucose uptake, cAMP and cGMP release, left ventricular developed pressure and coronary flow in isolated mouse hearts (Ban et al .…”

Section: Glp‐1 Receptor Expressionmentioning

confidence: 99%

“…These inconsistencies in GLP-1R expression in the heart cannot be attributed to possible variations in different mice strains as both studies used C57Bl/6 background mice (Ban et al 2008, Richards et al 2014. However, earlier studies used RT-PCR, immunohistochemistry and Western blotting to detect GLP-1 expression in the heart (Ban et al 2008), and the specificity and accuracy of commercially available GLP-1R antibodies have been questioned (Aroor and Nistala, 2014). Nevertheless, GLP-1 administration increased glucose uptake, cAMP and cGMP release, left ventricular developed pressure and coronary flow in isolated mouse hearts (Ban et al 2008), raising the possibilities of functional GLP-1R expression in the myocardium, a second receptor for GLP-1 or a key role of GLP-1R expressed in the endocardium, arterioles and arteries in the heart in modulating cardiac function.…”

Section: Glp-1 Receptor Expressionmentioning

confidence: 99%

Mechanisms for the cardiovascular effects of glucagon-like peptide-1

Poudyal

2015

Acta Physiol

View full text Add to dashboard Cite

Over the past three decades, at least 10 hormones secreted by the enteroendocrine cells have been discovered, which directly affect the cardiovascular system through their innate receptors expressed in the heart and blood vessels or through a neural mechanism. Glucagon-like peptide-1 (GLP-1), an important incretin, is perhaps best studied of these gut-derived hormones with important cardiovascular effects. In this review, I have discussed the mechanism of GLP-1 release from the enteroendocrine L-cells and its physiological effects on the cardiovascular system. Current evidence suggests that GLP-1 has positive inotropic and chronotropic effects on the heart and may be important in preserving left ventricular structure and function by direct and indirect mechanisms. The direct effects of GLP-1 in the heart may be mediated through GLP-1R expressed in atria as well as arteries and arterioles in the left ventricle and mainly involve in the activation of multiple pro-survival kinases and enhanced energy utilization. There is also good evidence to support the involvement of a second, yet to be identified, GLP-1 receptor. Further, GLP-1(9-36)amide, which was previously thought to be the inactive metabolite of the active GLP-1(7-36)amide, may also have direct cardioprotective effects. GLP-1's action on GLP-1R expressed in the central nervous system, kidney, vasculature and the pancreas may indirectly contribute to its cardioprotective effects.

show abstract

Tissue-Specific Expression of GLP1R in Mice: Is the Problem of Antibody Nonspecificity Solved?

Cited by 11 publications

References 18 publications

Super-resolution microscopy compatible fluorescent probes reveal endogenous glucagon-like peptide-1 receptor distribution and dynamics

Super-resolution microscopy compatible fluorescent probes reveal endogenous glucagon-like peptide-1 receptor distribution and dynamics

Distribution and characterisation of Glucagon-like peptide-1 receptor expressing cells in the mouse brain

Mechanisms for the cardiovascular effects of glucagon-like peptide-1

Contact Info

Product

Resources

About