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

Cork SC, Richards JE, Holt MK, Gribble FM, Reimann F, Trapp S - Mol Metab (2015)

Bottom Line: Large numbers of eYFP or tdRFP immunoreactive cells were found in the circumventricular organs, amygdala, hypothalamic nuclei and the ventrolateral medulla.However, tdRFP positive neurons were also found in areas without preproglucagon-neuronal projections like hippocampus and cortex.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.

View Article: PubMed Central - PubMed

Affiliation: Centre for Cardiovascular and Metabolic Neuroscience, Department of Neuroscience, Physiology & Pharmacology, University College London, London, WC1E 6BT, UK.

ABSTRACT

Objective: Although 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.

Methods: Mice 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.

Results: Large 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.

Conclusions: This 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.

No MeSH data available.


Related in: MedlinePlus

Diagrams of coronal sections showing the distribution of GLP-1R expressing cell bodies in the brains of GLP-1R-Cre mice. Filled circles represent the presence of YFP- or RFP-immunoreactive somata. The density of the filled circles indicates the relative density of the RFP-positive somata in each brain region. Drawings are based on the Paxinos Mouse Brain Atlas and numerical values next to each section indicate the rostro-caudal position in relation to Bregma. Abbreviations: PrL, prelimbic cortex; FrA, frontal association cortex; MO, medial orbital cortex; EPI, external plexiform layer; LS, lateral septum; AcbC, nucleus accumbens core; AcbS, nucleus accumbens shell; MS, medial septum nucleus; Cl, claustrum; Pir, piriform cortex; Cpu, caudate putamen; BNST-D/V, bed nucleus of the stria terminalis dorsal/ventral; PO, preoptic area; MPO, medial preoptic area; MnPO, median preoptic area; SFO, subfornical organ; Re, reuniens thalamic nucleus; PVN, paraventricular nucleus; LH, lateral hypothalamus; ARC, arcuate nucleus; Rt, reticular nucleus; PVT, thalamic paraventricular nucleus; VPM, ventral posteromedial thalamic nucleus; CeA, central amygdala; BLA, basolateral amygdala; DMH, dorsomedial hypothalamus; VMH, ventromedial hypothalamus; PAG, periaqueductal grey area; PH, posterior hypothalamus; PSTh, parasubthalamic nucleus; MM, mammillary nucleus; DG, dentate gyrus; DLG, dorsolateral geniculate nucleus; PGMC, pregeniculate nucleus magnocellular part; SCol, superior colliculus; VTA, ventral tegmental area; MG, medial geniculate nucleus; AP, area postrema; NTS, nucleus tractus solitarius; XII, hypoglossal nucleus; Ro, nucleus of roller; IRt, intermediate reticular nucleus; MdD, dorsal medullary reticular nucleus; RVLM, rostral ventrolateral medulla.
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fig2: Diagrams of coronal sections showing the distribution of GLP-1R expressing cell bodies in the brains of GLP-1R-Cre mice. Filled circles represent the presence of YFP- or RFP-immunoreactive somata. The density of the filled circles indicates the relative density of the RFP-positive somata in each brain region. Drawings are based on the Paxinos Mouse Brain Atlas and numerical values next to each section indicate the rostro-caudal position in relation to Bregma. Abbreviations: PrL, prelimbic cortex; FrA, frontal association cortex; MO, medial orbital cortex; EPI, external plexiform layer; LS, lateral septum; AcbC, nucleus accumbens core; AcbS, nucleus accumbens shell; MS, medial septum nucleus; Cl, claustrum; Pir, piriform cortex; Cpu, caudate putamen; BNST-D/V, bed nucleus of the stria terminalis dorsal/ventral; PO, preoptic area; MPO, medial preoptic area; MnPO, median preoptic area; SFO, subfornical organ; Re, reuniens thalamic nucleus; PVN, paraventricular nucleus; LH, lateral hypothalamus; ARC, arcuate nucleus; Rt, reticular nucleus; PVT, thalamic paraventricular nucleus; VPM, ventral posteromedial thalamic nucleus; CeA, central amygdala; BLA, basolateral amygdala; DMH, dorsomedial hypothalamus; VMH, ventromedial hypothalamus; PAG, periaqueductal grey area; PH, posterior hypothalamus; PSTh, parasubthalamic nucleus; MM, mammillary nucleus; DG, dentate gyrus; DLG, dorsolateral geniculate nucleus; PGMC, pregeniculate nucleus magnocellular part; SCol, superior colliculus; VTA, ventral tegmental area; MG, medial geniculate nucleus; AP, area postrema; NTS, nucleus tractus solitarius; XII, hypoglossal nucleus; Ro, nucleus of roller; IRt, intermediate reticular nucleus; MdD, dorsal medullary reticular nucleus; RVLM, rostral ventrolateral medulla.

Mentions: A schematic representation of the distribution of GLP-1R neurons throughout the mouse brain is given in Figure 2 and the density of GLP-1R neurons in individual brain nuclei is quantified in Table 1. Within the lower brainstem, GLP-1R neurons were scattered towards the lateral aspects of the NTS and throughout the intermediate reticular nucleus. Scattered neurons were also observed in the rostral ventrolateral medulla, locus coeruleus and nucleus of Roller. Further rostral few neurons were observed in the geniculate nucleus, mammillary nucleus, pretectal nucleus, and superior colliculus.


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

Cork SC, Richards JE, Holt MK, Gribble FM, Reimann F, Trapp S - Mol Metab (2015)

Diagrams of coronal sections showing the distribution of GLP-1R expressing cell bodies in the brains of GLP-1R-Cre mice. Filled circles represent the presence of YFP- or RFP-immunoreactive somata. The density of the filled circles indicates the relative density of the RFP-positive somata in each brain region. Drawings are based on the Paxinos Mouse Brain Atlas and numerical values next to each section indicate the rostro-caudal position in relation to Bregma. Abbreviations: PrL, prelimbic cortex; FrA, frontal association cortex; MO, medial orbital cortex; EPI, external plexiform layer; LS, lateral septum; AcbC, nucleus accumbens core; AcbS, nucleus accumbens shell; MS, medial septum nucleus; Cl, claustrum; Pir, piriform cortex; Cpu, caudate putamen; BNST-D/V, bed nucleus of the stria terminalis dorsal/ventral; PO, preoptic area; MPO, medial preoptic area; MnPO, median preoptic area; SFO, subfornical organ; Re, reuniens thalamic nucleus; PVN, paraventricular nucleus; LH, lateral hypothalamus; ARC, arcuate nucleus; Rt, reticular nucleus; PVT, thalamic paraventricular nucleus; VPM, ventral posteromedial thalamic nucleus; CeA, central amygdala; BLA, basolateral amygdala; DMH, dorsomedial hypothalamus; VMH, ventromedial hypothalamus; PAG, periaqueductal grey area; PH, posterior hypothalamus; PSTh, parasubthalamic nucleus; MM, mammillary nucleus; DG, dentate gyrus; DLG, dorsolateral geniculate nucleus; PGMC, pregeniculate nucleus magnocellular part; SCol, superior colliculus; VTA, ventral tegmental area; MG, medial geniculate nucleus; AP, area postrema; NTS, nucleus tractus solitarius; XII, hypoglossal nucleus; Ro, nucleus of roller; IRt, intermediate reticular nucleus; MdD, dorsal medullary reticular nucleus; RVLM, rostral ventrolateral medulla.
© Copyright Policy - CC BY
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4588458&req=5

fig2: Diagrams of coronal sections showing the distribution of GLP-1R expressing cell bodies in the brains of GLP-1R-Cre mice. Filled circles represent the presence of YFP- or RFP-immunoreactive somata. The density of the filled circles indicates the relative density of the RFP-positive somata in each brain region. Drawings are based on the Paxinos Mouse Brain Atlas and numerical values next to each section indicate the rostro-caudal position in relation to Bregma. Abbreviations: PrL, prelimbic cortex; FrA, frontal association cortex; MO, medial orbital cortex; EPI, external plexiform layer; LS, lateral septum; AcbC, nucleus accumbens core; AcbS, nucleus accumbens shell; MS, medial septum nucleus; Cl, claustrum; Pir, piriform cortex; Cpu, caudate putamen; BNST-D/V, bed nucleus of the stria terminalis dorsal/ventral; PO, preoptic area; MPO, medial preoptic area; MnPO, median preoptic area; SFO, subfornical organ; Re, reuniens thalamic nucleus; PVN, paraventricular nucleus; LH, lateral hypothalamus; ARC, arcuate nucleus; Rt, reticular nucleus; PVT, thalamic paraventricular nucleus; VPM, ventral posteromedial thalamic nucleus; CeA, central amygdala; BLA, basolateral amygdala; DMH, dorsomedial hypothalamus; VMH, ventromedial hypothalamus; PAG, periaqueductal grey area; PH, posterior hypothalamus; PSTh, parasubthalamic nucleus; MM, mammillary nucleus; DG, dentate gyrus; DLG, dorsolateral geniculate nucleus; PGMC, pregeniculate nucleus magnocellular part; SCol, superior colliculus; VTA, ventral tegmental area; MG, medial geniculate nucleus; AP, area postrema; NTS, nucleus tractus solitarius; XII, hypoglossal nucleus; Ro, nucleus of roller; IRt, intermediate reticular nucleus; MdD, dorsal medullary reticular nucleus; RVLM, rostral ventrolateral medulla.
Mentions: A schematic representation of the distribution of GLP-1R neurons throughout the mouse brain is given in Figure 2 and the density of GLP-1R neurons in individual brain nuclei is quantified in Table 1. Within the lower brainstem, GLP-1R neurons were scattered towards the lateral aspects of the NTS and throughout the intermediate reticular nucleus. Scattered neurons were also observed in the rostral ventrolateral medulla, locus coeruleus and nucleus of Roller. Further rostral few neurons were observed in the geniculate nucleus, mammillary nucleus, pretectal nucleus, and superior colliculus.

Bottom Line: Large numbers of eYFP or tdRFP immunoreactive cells were found in the circumventricular organs, amygdala, hypothalamic nuclei and the ventrolateral medulla.However, tdRFP positive neurons were also found in areas without preproglucagon-neuronal projections like hippocampus and cortex.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.

View Article: PubMed Central - PubMed

Affiliation: Centre for Cardiovascular and Metabolic Neuroscience, Department of Neuroscience, Physiology & Pharmacology, University College London, London, WC1E 6BT, UK.

ABSTRACT

Objective: Although 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.

Methods: Mice 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.

Results: Large 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.

Conclusions: This 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.

No MeSH data available.


Related in: MedlinePlus