Compartmentalization of endocannabinoids into lipid rafts in a microglial cell line devoid of caveolin-1

Neta Rimmerman*, Heather B. Bradshaw, Ewa Kozela, Rivka Levy, Ana Juknat, Zvi Vogel

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

26 Scopus citations

Abstract

BACKGROUND AND PURPOSE: N-acyl ethanolamines (NAEs) and 2-arachidonoyl glycerol (2-AG) are endogenous cannabinoids and along with related lipids are synthesized on demand from membrane phospholipids. Here, we have studied the compartmentalization of NAEs and 2-AG into lipid raft fractions isolated from the caveolin-1-lacking microglial cell line BV-2, following vehicle or cannabidiol (CBD) treatment. Results were compared with those from the caveolin-1-positive F-11 cell line. EXPERIMENTAL APPROACH: BV-2 cells were incubated with CBD or vehicle. Cells were fractionated using a detergent-free continuous OptiPrep density gradient. Lipids in fractions were quantified using HPLC/MS/MS. Proteins were measured using Western blot. KEY RESULTS: BV-2 cells were devoid of caveolin-1. Lipid rafts were isolated from BV-2 cells as confirmed by co-localization with flotillin-1 and sphingomyelin. Small amounts of cannabinoid CB1 receptors were found in lipid raft fractions. After incubation with CBD, levels and distribution in lipid rafts of 2-AG, N-arachidonoyl ethanolamine (AEA), and N-oleoyl ethanolamine (OEA) were not changed. Conversely, the levels of the saturated N-stearoyl ethanolamine (SEA) and N-palmitoyl ethanolamine (PEA) were elevated in lipid raft fractions. In whole cells with growth medium, CBD treatment increased AEA and OEA time-dependently, while levels of 2-AG, PEA and SEA did not change. CONCLUSIONS AND IMPLICATIONS: Whereas levels of 2-AG were not affected by CBD treatment, the distribution and levels of NAEs showed significant changes. Among the NAEs, the degree of acyl chain saturation predicted the compartmentalization after CBD treatment suggesting a shift in cell signalling activity. LINKED ARTICLES: This article is part of a themed section on Cannabinoids in Biology and Medicine. To view the other articles in this section visit http://dx.doi.org/10.1111/bph. 2012.165.issue-8. To view Part I of Cannabinoids in Biology and Medicine visit http://dx.doi.org/10.1111/bph.2011.163.issue-7.

Original languageAmerican English
Pages (from-to)2436-2449
Number of pages14
JournalBritish Journal of Pharmacology
Volume165
Issue number8
DOIs
StatePublished - Apr 2012
Externally publishedYes

Keywords

  • CB receptor
  • Cannabidiol
  • Cannabinoid
  • Caveolin-1
  • Endocannabinoid
  • Flotillin-1
  • Lipid raft
  • Mass spectrometry
  • Membrane raft
  • Microglia

Fingerprint

Dive into the research topics of 'Compartmentalization of endocannabinoids into lipid rafts in a microglial cell line devoid of caveolin-1'. Together they form a unique fingerprint.

Cite this