Binding, degradation and apoptotic activity of stearoylethanolamide in rat C6 glioma cells

Mauro Maccarrone, Riccardo Pauselli, Marianna Di Rienzo, Alessandro Finazzi-Agrò

Research output: Contribution to journalArticle

Abstract

Stearoylethanolamide (SEA) is present in human, rat and mouse brain in amounts comparable with those of the endocannabinoid anandamide (arachidonoylethanolamide; AEA). Yet, the biological activity of SEA has never been investigated. We synthesized unlabelled and radiolabelled SEA to investigate its binding, degradation and biological activity in rat C6 glioma cells. We report that SEA binds to a specific site distinct from known cannabinoid or vanilloid receptors, and that AEA and capsazepine partly (approx. 50%) antagonized this binding. Treatment of C6 cells with SEA inhibits cellular nitric oxide synthase and does not affect adenylate cyclase, whereas treatment with cannabinoid type 1 agonist 2-arachidonoylglycerol activates the former enzyme and inhibits the latter. C6 cells also have a specific SEA membrane transporter, which is inhibited by NO, and a fatty acid amide hydrolase capable of cleaving SEA. In these cells, SEA shows pro-apoptotic activity, due to elevation of intracellular calcium, activation of the arachidonate cascade and mitochondrial uncoupling. NO further enhances SEA-induced apoptosis. Moreover, the cannabinoid type 1 receptor-mediated decrease in cAMP induced by AEA in C6 cells is potentiated by SEA, suggesting that this compound also has an 'entourage' effect. Taken together, this study shows that SEA is an endocannabinoid-like compound which binds to and is transported by new components of the endocannabinoid system. It seems noteworthy that degradation and pro-apoptotic activity of SEA are regulated by NO in a way opposite to that reported for AEA.

Original languageEnglish
Pages (from-to)137-144
Number of pages8
JournalBiochemical Journal
Volume366
Issue number1
DOIs
Publication statusPublished - Aug 15 2002

Fingerprint

Glioma
Rats
Degradation
Endocannabinoids
Cannabinoid Receptors
Cannabinoids
Bioactivity
stearoylethanolamide
Cannabinoid Receptor Agonists
TRPV Cation Channels
Membrane Transport Proteins
Adenylyl Cyclases
Nitric Oxide Synthase
Brain
Chemical activation
Apoptosis
Calcium

Keywords

  • Arachidonate cascade
  • Calcium
  • Endocannabinoids
  • Mitochondria
  • Nitric oxide

ASJC Scopus subject areas

  • Biochemistry

Cite this

Binding, degradation and apoptotic activity of stearoylethanolamide in rat C6 glioma cells. / Maccarrone, Mauro; Pauselli, Riccardo; Di Rienzo, Marianna; Finazzi-Agrò, Alessandro.

In: Biochemical Journal, Vol. 366, No. 1, 15.08.2002, p. 137-144.

Research output: Contribution to journalArticle

Maccarrone, Mauro ; Pauselli, Riccardo ; Di Rienzo, Marianna ; Finazzi-Agrò, Alessandro. / Binding, degradation and apoptotic activity of stearoylethanolamide in rat C6 glioma cells. In: Biochemical Journal. 2002 ; Vol. 366, No. 1. pp. 137-144.
@article{e69b55020af64d1282c00e71b90926d3,
title = "Binding, degradation and apoptotic activity of stearoylethanolamide in rat C6 glioma cells",
abstract = "Stearoylethanolamide (SEA) is present in human, rat and mouse brain in amounts comparable with those of the endocannabinoid anandamide (arachidonoylethanolamide; AEA). Yet, the biological activity of SEA has never been investigated. We synthesized unlabelled and radiolabelled SEA to investigate its binding, degradation and biological activity in rat C6 glioma cells. We report that SEA binds to a specific site distinct from known cannabinoid or vanilloid receptors, and that AEA and capsazepine partly (approx. 50{\%}) antagonized this binding. Treatment of C6 cells with SEA inhibits cellular nitric oxide synthase and does not affect adenylate cyclase, whereas treatment with cannabinoid type 1 agonist 2-arachidonoylglycerol activates the former enzyme and inhibits the latter. C6 cells also have a specific SEA membrane transporter, which is inhibited by NO, and a fatty acid amide hydrolase capable of cleaving SEA. In these cells, SEA shows pro-apoptotic activity, due to elevation of intracellular calcium, activation of the arachidonate cascade and mitochondrial uncoupling. NO further enhances SEA-induced apoptosis. Moreover, the cannabinoid type 1 receptor-mediated decrease in cAMP induced by AEA in C6 cells is potentiated by SEA, suggesting that this compound also has an 'entourage' effect. Taken together, this study shows that SEA is an endocannabinoid-like compound which binds to and is transported by new components of the endocannabinoid system. It seems noteworthy that degradation and pro-apoptotic activity of SEA are regulated by NO in a way opposite to that reported for AEA.",
keywords = "Arachidonate cascade, Calcium, Endocannabinoids, Mitochondria, Nitric oxide",
author = "Mauro Maccarrone and Riccardo Pauselli and {Di Rienzo}, Marianna and Alessandro Finazzi-Agr{\`o}",
year = "2002",
month = "8",
day = "15",
doi = "10.1042/BJ20020438",
language = "English",
volume = "366",
pages = "137--144",
journal = "Biochemical Journal",
issn = "0264-6021",
publisher = "Portland Press Ltd.",
number = "1",

}

TY - JOUR

T1 - Binding, degradation and apoptotic activity of stearoylethanolamide in rat C6 glioma cells

AU - Maccarrone, Mauro

AU - Pauselli, Riccardo

AU - Di Rienzo, Marianna

AU - Finazzi-Agrò, Alessandro

PY - 2002/8/15

Y1 - 2002/8/15

N2 - Stearoylethanolamide (SEA) is present in human, rat and mouse brain in amounts comparable with those of the endocannabinoid anandamide (arachidonoylethanolamide; AEA). Yet, the biological activity of SEA has never been investigated. We synthesized unlabelled and radiolabelled SEA to investigate its binding, degradation and biological activity in rat C6 glioma cells. We report that SEA binds to a specific site distinct from known cannabinoid or vanilloid receptors, and that AEA and capsazepine partly (approx. 50%) antagonized this binding. Treatment of C6 cells with SEA inhibits cellular nitric oxide synthase and does not affect adenylate cyclase, whereas treatment with cannabinoid type 1 agonist 2-arachidonoylglycerol activates the former enzyme and inhibits the latter. C6 cells also have a specific SEA membrane transporter, which is inhibited by NO, and a fatty acid amide hydrolase capable of cleaving SEA. In these cells, SEA shows pro-apoptotic activity, due to elevation of intracellular calcium, activation of the arachidonate cascade and mitochondrial uncoupling. NO further enhances SEA-induced apoptosis. Moreover, the cannabinoid type 1 receptor-mediated decrease in cAMP induced by AEA in C6 cells is potentiated by SEA, suggesting that this compound also has an 'entourage' effect. Taken together, this study shows that SEA is an endocannabinoid-like compound which binds to and is transported by new components of the endocannabinoid system. It seems noteworthy that degradation and pro-apoptotic activity of SEA are regulated by NO in a way opposite to that reported for AEA.

AB - Stearoylethanolamide (SEA) is present in human, rat and mouse brain in amounts comparable with those of the endocannabinoid anandamide (arachidonoylethanolamide; AEA). Yet, the biological activity of SEA has never been investigated. We synthesized unlabelled and radiolabelled SEA to investigate its binding, degradation and biological activity in rat C6 glioma cells. We report that SEA binds to a specific site distinct from known cannabinoid or vanilloid receptors, and that AEA and capsazepine partly (approx. 50%) antagonized this binding. Treatment of C6 cells with SEA inhibits cellular nitric oxide synthase and does not affect adenylate cyclase, whereas treatment with cannabinoid type 1 agonist 2-arachidonoylglycerol activates the former enzyme and inhibits the latter. C6 cells also have a specific SEA membrane transporter, which is inhibited by NO, and a fatty acid amide hydrolase capable of cleaving SEA. In these cells, SEA shows pro-apoptotic activity, due to elevation of intracellular calcium, activation of the arachidonate cascade and mitochondrial uncoupling. NO further enhances SEA-induced apoptosis. Moreover, the cannabinoid type 1 receptor-mediated decrease in cAMP induced by AEA in C6 cells is potentiated by SEA, suggesting that this compound also has an 'entourage' effect. Taken together, this study shows that SEA is an endocannabinoid-like compound which binds to and is transported by new components of the endocannabinoid system. It seems noteworthy that degradation and pro-apoptotic activity of SEA are regulated by NO in a way opposite to that reported for AEA.

KW - Arachidonate cascade

KW - Calcium

KW - Endocannabinoids

KW - Mitochondria

KW - Nitric oxide

UR - http://www.scopus.com/inward/record.url?scp=0037103724&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037103724&partnerID=8YFLogxK

U2 - 10.1042/BJ20020438

DO - 10.1042/BJ20020438

M3 - Article

C2 - 12010121

AN - SCOPUS:0037103724

VL - 366

SP - 137

EP - 144

JO - Biochemical Journal

JF - Biochemical Journal

SN - 0264-6021

IS - 1

ER -