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Compartmentalization of endocannabinoids into lipid rafts in a microglial cell line devoid of caveolin-1.
Br J Pharmacol. 2012 Apr; 165(8):2436-49.BJ

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 CB(1) 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.

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Authors+Show Affiliations

Rimmerman N
Department of Neurobiology, Weizmann Institute of Science, Rehovot, Israel. Neta.rimmerman@weizmann.ac.il
Bradshaw HB
No affiliation info available
Kozela E
No affiliation info available
Levy R
No affiliation info available
Juknat A
No affiliation info available
Vogel Z
No affiliation info available

MeSH

AnimalsCannabidiolCannabinoid Receptor ModulatorsCaveolin 1Cell LineEndocannabinoidsMembrane MicrodomainsMiceMicrogliaRatsReceptor, Cannabinoid, CB1Receptor, Cannabinoid, CB2

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

21449981

Citation

Rimmerman, Neta, et al. "Compartmentalization of Endocannabinoids Into Lipid Rafts in a Microglial Cell Line Devoid of Caveolin-1." British Journal of Pharmacology, vol. 165, no. 8, 2012, pp. 2436-49.
Rimmerman N, Bradshaw HB, Kozela E, et al. Compartmentalization of endocannabinoids into lipid rafts in a microglial cell line devoid of caveolin-1. Br J Pharmacol. 2012;165(8):2436-49.
Rimmerman, N., Bradshaw, H. B., Kozela, E., Levy, R., Juknat, A., & Vogel, Z. (2012). Compartmentalization of endocannabinoids into lipid rafts in a microglial cell line devoid of caveolin-1. British Journal of Pharmacology, 165(8), 2436-49. https://doi.org/10.1111/j.1476-5381.2011.01380.x
Rimmerman N, et al. Compartmentalization of Endocannabinoids Into Lipid Rafts in a Microglial Cell Line Devoid of Caveolin-1. Br J Pharmacol. 2012;165(8):2436-49. PubMed PMID: 21449981.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Compartmentalization of endocannabinoids into lipid rafts in a microglial cell line devoid of caveolin-1. AU - Rimmerman,Neta, AU - Bradshaw,Heather B, AU - Kozela,Ewa, AU - Levy,Rivka, AU - Juknat,Ana, AU - Vogel,Zvi, PY - 2011/4/1/entrez PY - 2011/4/1/pubmed PY - 2012/7/28/medline SP - 2436 EP - 49 JF - British journal of pharmacology JO - Br J Pharmacol VL - 165 IS - 8 N2 - 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 CB(1) 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. SN - 1476-5381 UR - https://www.unboundmedicine.com/medline/citation/21449981/Compartmentalization_of_endocannabinoids_into_lipid_rafts_in_a_microglial_cell_line_devoid_of_caveolin_1_ L2 - https://doi.org/10.1111/j.1476-5381.2011.01380.x DB - PRIME DP - Unbound Medicine ER -