Serotonergic modulation of the activity of GLP-1 producing neurons in the nucleus of the solitary tract in mouse

Marie K. Holt; Ida J. Llewellyn-Smith; Frank Reimann; Fiona M. Gribble; Stefan Trapp

doi:10.1016/j.molmet.2017.06.002

Serotonergic modulation of the activity of GLP-1 producing neurons in the nucleus of the solitary tract in mouse

Marie K. Holt, Ida J. Llewellyn-Smith, Frank Reimann, Fiona M. Gribble, Stefan Trapp

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Abstract

Objective

Glucagon-like peptide-1 (GLP-1) and 5-HT are potent regulators of food intake within the brain. GLP-1 is expressed by preproglucagon (PPG) neurons in the nucleus tractus solitarius (NTS). We have previously shown that PPG neurons innervate 5-HT neurons in the ventral brainstem. Here, we investigate whether PPG neurons receive serotonergic input and respond to 5-HT.

Methods

We employed immunohistochemistry to reveal serotonergic innervation of PPG neurons. We investigated the responsiveness of PPG neurons to 5-HT using in vitro Ca²⁺ imaging in brainstem slices from transgenic mice expressing the Ca²⁺ indicator, GCaMP3, in PPG neurons, and cell-attached patch-clamp recordings.

Results

Close appositions from 5-HT-immunoreactive axons occurred on many PPG neurons. Application of 20 μM 5-HT produced robust Ca²⁺ responses in NTS PPG dendrites but little change in somata. Dendritic Ca²⁺ spikes were concentration-dependent (2, 20, and 200 μM) and unaffected by blockade of glutamatergic transmission, suggesting 5-HT receptors on PPG neurons. Neither activation nor blockade of 5-HT₃ receptors affected [Ca²⁺]_i. In contrast, inhibition of 5-HT₂ receptors attenuated increases in intracellular Ca²⁺ and 5-HT_2C receptor activation produced Ca²⁺ spikes. Patch-clamp recordings revealed that 44% of cells decreased their firing rate under 5-HT, an effect blocked by 5-HT_1A receptor antagonism.

Conclusions PPG neurons respond directly to 5-HT with a 5-HT_2C receptor-dependent increase in dendritic [Ca²⁺]_i. Electrical responses to 5-HT revealed additional inhibitory effects due to somatic 5-HT_1A receptors. Reciprocal innervation between 5-HT and PPG neurons suggests that the coordinated activity of these brainstem neurons may play a role in the regulation of food intake.

Original language	English
Pages (from-to)	909-921
Number of pages	13
Journal	Molecular Metabolism
Volume	6
Issue number	8
DOIs	https://doi.org/10.1016/j.molmet.2017.06.002
Publication status	Published - 1 Aug 2017

Bibliographical note

Keywords

Dendritic calcium
GCaMP
NTS
Preproglucagon
Serotonin

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title = "Serotonergic modulation of the activity of GLP-1 producing neurons in the nucleus of the solitary tract in mouse",

abstract = "Objective Glucagon-like peptide-1 (GLP-1) and 5-HT are potent regulators of food intake within the brain. GLP-1 is expressed by preproglucagon (PPG) neurons in the nucleus tractus solitarius (NTS). We have previously shown that PPG neurons innervate 5-HT neurons in the ventral brainstem. Here, we investigate whether PPG neurons receive serotonergic input and respond to 5-HT. Methods We employed immunohistochemistry to reveal serotonergic innervation of PPG neurons. We investigated the responsiveness of PPG neurons to 5-HT using in vitro Ca2+ imaging in brainstem slices from transgenic mice expressing the Ca2+ indicator, GCaMP3, in PPG neurons, and cell-attached patch-clamp recordings.Results Close appositions from 5-HT-immunoreactive axons occurred on many PPG neurons. Application of 20 μM 5-HT produced robust Ca2+ responses in NTS PPG dendrites but little change in somata. Dendritic Ca2+ spikes were concentration-dependent (2, 20, and 200 μM) and unaffected by blockade of glutamatergic transmission, suggesting 5-HT receptors on PPG neurons. Neither activation nor blockade of 5-HT3 receptors affected [Ca2+]i. In contrast, inhibition of 5-HT2 receptors attenuated increases in intracellular Ca2+ and 5-HT2C receptor activation produced Ca2+ spikes. Patch-clamp recordings revealed that 44% of cells decreased their firing rate under 5-HT, an effect blocked by 5-HT1A receptor antagonism. Conclusions PPG neurons respond directly to 5-HT with a 5-HT2C receptor-dependent increase in dendritic [Ca2+]i. Electrical responses to 5-HT revealed additional inhibitory effects due to somatic 5-HT1A receptors. Reciprocal innervation between 5-HT and PPG neurons suggests that the coordinated activity of these brainstem neurons may play a role in the regulation of food intake.",

keywords = "Dendritic calcium, GCaMP, NTS, Preproglucagon, Serotonin",

author = "Holt, {Marie K.} and Llewellyn-Smith, {Ida J.} and Frank Reimann and Gribble, {Fiona M.} and Stefan Trapp",

year = "2017",

month = aug,

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doi = "10.1016/j.molmet.2017.06.002",

language = "English",

volume = "6",

pages = "909--921",

journal = "Molecular Metabolism",

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T1 - Serotonergic modulation of the activity of GLP-1 producing neurons in the nucleus of the solitary tract in mouse

AU - Holt, Marie K.

AU - Llewellyn-Smith, Ida J.

AU - Reimann, Frank

AU - Gribble, Fiona M.

AU - Trapp, Stefan

PY - 2017/8/1

Y1 - 2017/8/1

N2 - Objective Glucagon-like peptide-1 (GLP-1) and 5-HT are potent regulators of food intake within the brain. GLP-1 is expressed by preproglucagon (PPG) neurons in the nucleus tractus solitarius (NTS). We have previously shown that PPG neurons innervate 5-HT neurons in the ventral brainstem. Here, we investigate whether PPG neurons receive serotonergic input and respond to 5-HT. Methods We employed immunohistochemistry to reveal serotonergic innervation of PPG neurons. We investigated the responsiveness of PPG neurons to 5-HT using in vitro Ca2+ imaging in brainstem slices from transgenic mice expressing the Ca2+ indicator, GCaMP3, in PPG neurons, and cell-attached patch-clamp recordings.Results Close appositions from 5-HT-immunoreactive axons occurred on many PPG neurons. Application of 20 μM 5-HT produced robust Ca2+ responses in NTS PPG dendrites but little change in somata. Dendritic Ca2+ spikes were concentration-dependent (2, 20, and 200 μM) and unaffected by blockade of glutamatergic transmission, suggesting 5-HT receptors on PPG neurons. Neither activation nor blockade of 5-HT3 receptors affected [Ca2+]i. In contrast, inhibition of 5-HT2 receptors attenuated increases in intracellular Ca2+ and 5-HT2C receptor activation produced Ca2+ spikes. Patch-clamp recordings revealed that 44% of cells decreased their firing rate under 5-HT, an effect blocked by 5-HT1A receptor antagonism. Conclusions PPG neurons respond directly to 5-HT with a 5-HT2C receptor-dependent increase in dendritic [Ca2+]i. Electrical responses to 5-HT revealed additional inhibitory effects due to somatic 5-HT1A receptors. Reciprocal innervation between 5-HT and PPG neurons suggests that the coordinated activity of these brainstem neurons may play a role in the regulation of food intake.

AB - Objective Glucagon-like peptide-1 (GLP-1) and 5-HT are potent regulators of food intake within the brain. GLP-1 is expressed by preproglucagon (PPG) neurons in the nucleus tractus solitarius (NTS). We have previously shown that PPG neurons innervate 5-HT neurons in the ventral brainstem. Here, we investigate whether PPG neurons receive serotonergic input and respond to 5-HT. Methods We employed immunohistochemistry to reveal serotonergic innervation of PPG neurons. We investigated the responsiveness of PPG neurons to 5-HT using in vitro Ca2+ imaging in brainstem slices from transgenic mice expressing the Ca2+ indicator, GCaMP3, in PPG neurons, and cell-attached patch-clamp recordings.Results Close appositions from 5-HT-immunoreactive axons occurred on many PPG neurons. Application of 20 μM 5-HT produced robust Ca2+ responses in NTS PPG dendrites but little change in somata. Dendritic Ca2+ spikes were concentration-dependent (2, 20, and 200 μM) and unaffected by blockade of glutamatergic transmission, suggesting 5-HT receptors on PPG neurons. Neither activation nor blockade of 5-HT3 receptors affected [Ca2+]i. In contrast, inhibition of 5-HT2 receptors attenuated increases in intracellular Ca2+ and 5-HT2C receptor activation produced Ca2+ spikes. Patch-clamp recordings revealed that 44% of cells decreased their firing rate under 5-HT, an effect blocked by 5-HT1A receptor antagonism. Conclusions PPG neurons respond directly to 5-HT with a 5-HT2C receptor-dependent increase in dendritic [Ca2+]i. Electrical responses to 5-HT revealed additional inhibitory effects due to somatic 5-HT1A receptors. Reciprocal innervation between 5-HT and PPG neurons suggests that the coordinated activity of these brainstem neurons may play a role in the regulation of food intake.

KW - Dendritic calcium

KW - GCaMP

KW - NTS

KW - Preproglucagon

KW - Serotonin

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DO - 10.1016/j.molmet.2017.06.002

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AN - SCOPUS:85020931634

SN - 2212-8778

VL - 6

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JO - Molecular Metabolism

JF - Molecular Metabolism

IS - 8

ER -

Serotonergic modulation of the activity of GLP-1 producing neurons in the nucleus of the solitary tract in mouse

Abstract

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Keywords

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