Prolonged Activation of the Htr2b Serotonin Receptor Impairs Glucose Stimulated Insulin Secretion and Mitochondrial Function in MIN6 Cells

Luis Rodrigo Cataldo; María L. Mizgier; Roberto Bravo Sagua; Fabián Jaña; César Cárdenas; Paola Llanos; Dolores Busso; Pablo Olmos; José E. Galgani; José L. Santos; Víctor A. Cortés

doi:10.1371/journal.pone.0170213

Prolonged Activation of the Htr2b Serotonin Receptor Impairs Glucose Stimulated Insulin Secretion and Mitochondrial Function in MIN6 Cells

Luis Rodrigo Cataldo, María L. Mizgier, Roberto Bravo Sagua, Fabián Jaña, César Cárdenas, Paola Llanos, Dolores Busso, Pablo Olmos, José E. Galgani, José L. Santos, Víctor A. Cortés

Research output: Contribution to journal › Article › peer-review

23 Scopus citations

Abstract

Aims Pancreatic β-cells synthesize and release serotonin (5 hydroxytryptamine, 5HT); however, the role of 5HT receptors on glucose stimulated insulin secretion (GSIS) and the mechanisms mediating this function is not fully understood. The aims of this study were to determine the expression profile of 5HT receptors in murine MIN6 β-cells and to examine the effects of pharmacological activation of 5HT receptor Htr2b on GSIS and mitochondrial function. Materials and Methods mRNA levels of 5HT receptors in MIN6 cells were quantified by RT qPCR. GSIS was assessed in MIN6 cells in response to global serotonergic activation with 5HT and pharmacological Htr2b activation or inhibition with BW723C86 or SB204741, respectively. In response to Htr2b activation also was evaluated the mRNA and protein levels of PGC1α and PPARy by RT-qPCR and western blotting and mitochondrial function by oxygen consumption rate (OCR) and ATP cellular content. Results We found that mRNA levels of most 5HT receptors were either very low or undetectable in MIN6 cells. By contrast, Htr2b mRNA was present at moderate levels in these cells. Preincubation (6 h) of MIN6 cells with 5HT or BW723C86 reduced GSIS and the effect of 5HT was prevented by SB204741. Preincubation with BW723C86 increased PGC1α and PPARy mRNA and protein levels and decreased mitochondrial respiration and ATP content in MIN6 cells. Conclusions Our results indicate that prolonged Htr2b activation in murine β-cells decreases glucosestimulated insulin secretion and mitochondrial activity by mechanisms likely dependent on enhanced PGC1α/PPARy expression.

Original language	English
Article number	e0170213
Journal	PLoS ONE
Volume	12
Issue number	1
DOIs	https://doi.org/10.1371/journal.pone.0170213
State	Published - Jan 2017
Externally published	Yes

Bibliographical note

Funding Information:
This research was funded by "Fondo Nacional de Desarrollo Cient?fico y Tecnol?gico" grants; 1141134 (VC), 1120586 (JLS), 1120443 (CC), 1160332 (CC), 1130217 (JEG). "Fondo de financiamiento de Centros de Investigaci?n en ?reas Prioritarias" grant 15150012 (CC). "Comisi?n Nacional de Investigaci?n en Ciencia y Tecnolog?a"; doctoral scholarship 21140087 (LRC), postdoctoral fellowship 3160226 (RBS), postdoctoral fellowship 3140458 (FJ). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Publisher Copyright:
© 2017 Cataldo et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Keywords

Adenosine Triphosphate
Animals
Cell Line
Gene Expression Regulation
Glucose
Humans
Indoles
Insulin
Insulin-Secreting Cells
Islets of Langerhans
Mice
Mitochondria
Oxygen Consumption
Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha
PPAR gamma
Receptors, Serotonin
Serotonin
Thiophenes
Urea

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10.1371/journal.pone.0170213

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Cataldo, L. R., Mizgier, M. L., Sagua, R. B., Jaña, F., Cárdenas, C., Llanos, P., Busso, D., Olmos, P., Galgani, J. E., Santos, J. L., & Cortés, V. A. (2017). Prolonged Activation of the Htr2b Serotonin Receptor Impairs Glucose Stimulated Insulin Secretion and Mitochondrial Function in MIN6 Cells. PLoS ONE, 12(1), Article e0170213. https://doi.org/10.1371/journal.pone.0170213

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title = "Prolonged Activation of the Htr2b Serotonin Receptor Impairs Glucose Stimulated Insulin Secretion and Mitochondrial Function in MIN6 Cells",

abstract = "Aims Pancreatic β-cells synthesize and release serotonin (5 hydroxytryptamine, 5HT); however, the role of 5HT receptors on glucose stimulated insulin secretion (GSIS) and the mechanisms mediating this function is not fully understood. The aims of this study were to determine the expression profile of 5HT receptors in murine MIN6 β-cells and to examine the effects of pharmacological activation of 5HT receptor Htr2b on GSIS and mitochondrial function. Materials and Methods mRNA levels of 5HT receptors in MIN6 cells were quantified by RT qPCR. GSIS was assessed in MIN6 cells in response to global serotonergic activation with 5HT and pharmacological Htr2b activation or inhibition with BW723C86 or SB204741, respectively. In response to Htr2b activation also was evaluated the mRNA and protein levels of PGC1α and PPARy by RT-qPCR and western blotting and mitochondrial function by oxygen consumption rate (OCR) and ATP cellular content. Results We found that mRNA levels of most 5HT receptors were either very low or undetectable in MIN6 cells. By contrast, Htr2b mRNA was present at moderate levels in these cells. Preincubation (6 h) of MIN6 cells with 5HT or BW723C86 reduced GSIS and the effect of 5HT was prevented by SB204741. Preincubation with BW723C86 increased PGC1α and PPARy mRNA and protein levels and decreased mitochondrial respiration and ATP content in MIN6 cells. Conclusions Our results indicate that prolonged Htr2b activation in murine β-cells decreases glucosestimulated insulin secretion and mitochondrial activity by mechanisms likely dependent on enhanced PGC1α/PPARy expression.",

keywords = "Adenosine Triphosphate, Animals, Cell Line, Gene Expression Regulation, Glucose, Humans, Indoles, Insulin, Insulin-Secreting Cells, Islets of Langerhans, Mice, Mitochondria, Oxygen Consumption, Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha, PPAR gamma, Receptors, Serotonin, Serotonin, Thiophenes, Urea",

author = "Cataldo, {Luis Rodrigo} and Mizgier, {Mar{\'i}a L.} and Sagua, {Roberto Bravo} and Fabi{\'a}n Ja{\~n}a and C{\'e}sar C{\'a}rdenas and Paola Llanos and Dolores Busso and Pablo Olmos and Galgani, {Jos{\'e} E.} and Santos, {Jos{\'e} L.} and Cort{\'e}s, {V{\'i}ctor A.}",

note = "Funding Information: This research was funded by {"}Fondo Nacional de Desarrollo Cient?fico y Tecnol?gico{"} grants; 1141134 (VC), 1120586 (JLS), 1120443 (CC), 1160332 (CC), 1130217 (JEG). {"}Fondo de financiamiento de Centros de Investigaci?n en ?reas Prioritarias{"} grant 15150012 (CC). {"}Comisi?n Nacional de Investigaci?n en Ciencia y Tecnolog?a{"}; doctoral scholarship 21140087 (LRC), postdoctoral fellowship 3160226 (RBS), postdoctoral fellowship 3140458 (FJ). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Publisher Copyright: {\textcopyright} 2017 Cataldo et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.",

year = "2017",

month = jan,

doi = "10.1371/journal.pone.0170213",

language = "English",

volume = "12",

journal = "PLoS ONE",

issn = "1932-6203",

publisher = "Public Library of Science",

number = "1",

}

TY - JOUR

T1 - Prolonged Activation of the Htr2b Serotonin Receptor Impairs Glucose Stimulated Insulin Secretion and Mitochondrial Function in MIN6 Cells

AU - Cataldo, Luis Rodrigo

AU - Mizgier, María L.

AU - Sagua, Roberto Bravo

AU - Jaña, Fabián

AU - Cárdenas, César

AU - Llanos, Paola

AU - Busso, Dolores

AU - Olmos, Pablo

AU - Galgani, José E.

AU - Santos, José L.

AU - Cortés, Víctor A.

N1 - Funding Information: This research was funded by "Fondo Nacional de Desarrollo Cient?fico y Tecnol?gico" grants; 1141134 (VC), 1120586 (JLS), 1120443 (CC), 1160332 (CC), 1130217 (JEG). "Fondo de financiamiento de Centros de Investigaci?n en ?reas Prioritarias" grant 15150012 (CC). "Comisi?n Nacional de Investigaci?n en Ciencia y Tecnolog?a"; doctoral scholarship 21140087 (LRC), postdoctoral fellowship 3160226 (RBS), postdoctoral fellowship 3140458 (FJ). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Publisher Copyright: © 2017 Cataldo et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

PY - 2017/1

Y1 - 2017/1

N2 - Aims Pancreatic β-cells synthesize and release serotonin (5 hydroxytryptamine, 5HT); however, the role of 5HT receptors on glucose stimulated insulin secretion (GSIS) and the mechanisms mediating this function is not fully understood. The aims of this study were to determine the expression profile of 5HT receptors in murine MIN6 β-cells and to examine the effects of pharmacological activation of 5HT receptor Htr2b on GSIS and mitochondrial function. Materials and Methods mRNA levels of 5HT receptors in MIN6 cells were quantified by RT qPCR. GSIS was assessed in MIN6 cells in response to global serotonergic activation with 5HT and pharmacological Htr2b activation or inhibition with BW723C86 or SB204741, respectively. In response to Htr2b activation also was evaluated the mRNA and protein levels of PGC1α and PPARy by RT-qPCR and western blotting and mitochondrial function by oxygen consumption rate (OCR) and ATP cellular content. Results We found that mRNA levels of most 5HT receptors were either very low or undetectable in MIN6 cells. By contrast, Htr2b mRNA was present at moderate levels in these cells. Preincubation (6 h) of MIN6 cells with 5HT or BW723C86 reduced GSIS and the effect of 5HT was prevented by SB204741. Preincubation with BW723C86 increased PGC1α and PPARy mRNA and protein levels and decreased mitochondrial respiration and ATP content in MIN6 cells. Conclusions Our results indicate that prolonged Htr2b activation in murine β-cells decreases glucosestimulated insulin secretion and mitochondrial activity by mechanisms likely dependent on enhanced PGC1α/PPARy expression.

AB - Aims Pancreatic β-cells synthesize and release serotonin (5 hydroxytryptamine, 5HT); however, the role of 5HT receptors on glucose stimulated insulin secretion (GSIS) and the mechanisms mediating this function is not fully understood. The aims of this study were to determine the expression profile of 5HT receptors in murine MIN6 β-cells and to examine the effects of pharmacological activation of 5HT receptor Htr2b on GSIS and mitochondrial function. Materials and Methods mRNA levels of 5HT receptors in MIN6 cells were quantified by RT qPCR. GSIS was assessed in MIN6 cells in response to global serotonergic activation with 5HT and pharmacological Htr2b activation or inhibition with BW723C86 or SB204741, respectively. In response to Htr2b activation also was evaluated the mRNA and protein levels of PGC1α and PPARy by RT-qPCR and western blotting and mitochondrial function by oxygen consumption rate (OCR) and ATP cellular content. Results We found that mRNA levels of most 5HT receptors were either very low or undetectable in MIN6 cells. By contrast, Htr2b mRNA was present at moderate levels in these cells. Preincubation (6 h) of MIN6 cells with 5HT or BW723C86 reduced GSIS and the effect of 5HT was prevented by SB204741. Preincubation with BW723C86 increased PGC1α and PPARy mRNA and protein levels and decreased mitochondrial respiration and ATP content in MIN6 cells. Conclusions Our results indicate that prolonged Htr2b activation in murine β-cells decreases glucosestimulated insulin secretion and mitochondrial activity by mechanisms likely dependent on enhanced PGC1α/PPARy expression.

KW - Adenosine Triphosphate

KW - Animals

KW - Cell Line

KW - Gene Expression Regulation

KW - Glucose

KW - Humans

KW - Indoles

KW - Insulin

KW - Insulin-Secreting Cells

KW - Islets of Langerhans

KW - Mice

KW - Mitochondria

KW - Oxygen Consumption

KW - Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha

KW - PPAR gamma

KW - Receptors, Serotonin

KW - Serotonin

KW - Thiophenes

KW - Urea

UR - http://www.scopus.com/inward/record.url?scp=85011024132&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0170213

DO - 10.1371/journal.pone.0170213

M3 - Article

C2 - 28129327

AN - SCOPUS:85011024132

SN - 1932-6203

VL - 12

JO - PLoS ONE

JF - PLoS ONE

IS - 1

M1 - e0170213

ER -

Prolonged Activation of the Htr2b Serotonin Receptor Impairs Glucose Stimulated Insulin Secretion and Mitochondrial Function in MIN6 Cells

Abstract

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Keywords

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