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Comparison of anandamide transport in FAAH wild-type and knockout neurons: Evidence for contributions by both FAAH and the CB1 receptor to anandamide uptake

Academic Article
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Overview

authors

  • Ortega-Gutierrez, S.
  • Hawkins, E. G.
  • Viso, A.
  • Lopez-Rodriguez, M. L.
  • Cravatt, Benjamin

publication date

  • June 2004

journal

  • Biochemistry  Journal

abstract

  • The cellular inactivation of the endogenous cannabinoid (endocannabinoid) anandamide (AEA) represents a controversial and intensely investigated subject. This process has been proposed to involve two proteins, a transporter that promotes the cellular uptake of AEA and fatty acid amide hydrolase (FAAH), which hydrolyzes AEA to arachidonic acid. However, whereas the role of FAAH in AEA metabolism is well-characterized, the identity of the putative AEA transporter remains enigmatic. Indeed, the indirect pharmacological evidence used to support the existence of an AEA transporter has been suggested also to be compatible with a model in which AEA uptake is driven by simple diffusion coupled to FAAH metabolism. Here, we have directly addressed the contribution of FAAH to AEA uptake by examining this process in neuronal preparations from FAAH(-/-) mice and in the presence of the uptake inhibitor UCM707. The results of these studies reveal that (i) care should be taken to avoid the presence of artifacts when studying the cellular uptake of lipophilic molecules like AEA, (ii) FAAH significantly contributes to AEA uptake, especially with longer incubation times, and (iii) a UCM707-sensitive protein(s) distinct from FAAH also participates in AEA uptake. Interestingly, the FAAH-independent component of AEA transport was significantly reduced by pretreatment of neurons with the cannabinoid receptor 1 (CB1) antagonist SR141716A. Collectively, these results indicate that the protein-dependent uptake of AEA is largely mediated by known constituents of the endocannabinoid system (FAAH and the CB1 receptor), although a partial contribution of an additional UCM707-sensitive protein is also suggested.

subject areas

  • Amidohydrolases
  • Animals
  • Arachidonic Acids
  • Biological Transport
  • Brain
  • Carrier Proteins
  • Cattle
  • Cells, Cultured
  • Embryo, Mammalian
  • Endocannabinoids
  • Furans
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Neurons
  • Piperidines
  • Polyunsaturated Alkamides
  • Pyrazoles
  • Receptor, Cannabinoid, CB1
  • Serum Albumin, Bovine
  • Tritium
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Identity

International Standard Serial Number (ISSN)

  • 0006-2960

Digital Object Identifier (DOI)

  • 10.1021/bi049395f

PubMed ID

  • 15209515
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Additional Document Info

start page

  • 8184

end page

  • 8190

volume

  • 43

issue

  • 25

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