Confocal microscopy and biochemical analysis reveal spatial and functional separation between anandamide uptake and hydrolysis in human keratinocytes

Oddi, Sergio; Bari, Monica; Battista, Natalia; Barsacchi, D; Cozzani, I.; Maccarrone, Mauro

doi:10.1007/s00018-004-4446-8

Cited by 63 publications

(53 citation statements)

References 60 publications

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“…1B and 1C). Wild-type FAAH-eGFP (COS7-FAAH-eGFP) localized to the ER in agreement with a previous study (38). TOM-⌬TMFAAH-eGFP and GRASP-⌬TMFAAH-eGFP were confined to the mitochondria and Golgi apparatus, respectively (Fig.…”

Section: Aea Cellular Uptake Is Rapid and Independent Of Subcellular supporting

confidence: 92%

“…We propose that these inhibitors compete with AEA for binding to commonly expressed intracellular carriers such as FABPs. For example, the inhibitor AM1172 reduced AEA accumulation in cultured neurons, whereas OMDM1 inhibited AEA uptake in HaCaT keratinocytes (38,61). Accordingly, neurons express FABP3 and FABP5 while HaCaT cells express FABP5 (27,33).…”

Section: Discussionmentioning

confidence: 98%

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Identification of intracellular carriers for the endocannabinoid anandamide

Kaczocha¹,

Glaser

Deutsch³

2009

Proc. Natl. Acad. Sci. U.S.A.

305

332

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The endocannabinoid anandamide (arachidonoyl ethanolamide, AEA) is an uncharged neuromodulatory lipid that, similar to many neurotransmitters, is inactivated through its cellular uptake and subsequent catabolism. AEA is hydrolyzed by fatty acid amide hydrolase (FAAH), an enzyme localized on the endoplasmic reticulum. In contrast to most neuromodulators, the hydrophilic cytosol poses a diffusional barrier for the efficient delivery of AEA to its site of catabolism. Therefore, AEA likely traverses the cytosol with the assistance of an intracellular carrier that increases its solubility and rate of diffusion. To study this process, AEA uptake and hydrolysis were examined in COS-7 cells expressing FAAH restricted to the endoplasmic reticulum, mitochondria, or the Golgi apparatus. AEA hydrolysis was detectable at the earliest measurable time point (

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Section: Aea Cellular Uptake Is Rapid and Independent Of Subcellular supporting

confidence: 92%

Section: Discussionmentioning

confidence: 98%

Identification of intracellular carriers for the endocannabinoid anandamide

Kaczocha¹,

Glaser

Deutsch³

2009

Proc. Natl. Acad. Sci. U.S.A.

305

332

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“…In the current models, the enzymes for the synthesis and degradation of endocannabinoids are thought to be located within the cell, so that the stimulation of cannabinoid receptors from the extracellular component by endocannabinoids requires endocannabinoids to cross the plasma membrane twice. While pharmacological and biochemical evidence points towards the existence of a specific anandamide transport protein using transporter inhibitors [23][24][25][26], no direct evidence for such a transporter has been provided. Recently developed drugs have been shown to inhibit anandamide transport without affecting FAAH activity [27].…”

Section: Aea Synthesis and Degradationmentioning

confidence: 99%

Aspects of endocannabinoid signaling in periimplantation biology

Sun

Dey

2008

Molecular and Cellular Endocrinology

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Physiological roles of endocannabinoids, a group of endogenously produced cannabinoid-like lipid molecules that activate G-protein coupled cannabinoid receptors, are being increasingly appreciated in female reproduction. Adverse effects of cannabinoids on female fertility have been suspected for decades; however, underlying molecular and genetic bases by which they exert these effects were not clearly understood. The discovery of cannabinoid receptors (CB1 and CB2), endocannabinoid ligands (anandamide and 2-acylglycerol) as well as their key synthetic and hydrolytic pathways has helped to better understand the roles of cannabinoid/endocannabinoid signaling in preimplantation embryo development, oviductal embryo transport, embryo implantation and postimplantation embryonic growth. This review focuses on various aspects of the endocannabinoid system in female fertility based on studies that used knockout mouse models. The information generated from studies in mice is likely to shed deeper insight into fertility regulation in women.

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“…For anandamide to be metabolized by FAAH, it first must be transported through the plasma membrane to the cellular compartments where FAAH is localized (32). Although the exact molecular nature of this transport process is still under debate, it is agreed that anandamide movement across the plasma membrane is saturable, rapid (33,34), temperature-dependent (33), and enantioselective (35), and it does not appear to require an ion gradient or chemical energy supply (ATP) (31).…”

mentioning

confidence: 99%

Identification of a high-affinity binding site involved in the transport of endocannabinoids

Moore

Nomikos²,

Dickason-Chesterfield³

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

151

153

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Phytocannabinoids, such as the principal bioactive component of marijuana, ⌬ 9 -tetrahydrocannabinol, have been used for thousands of years for medical and recreational purposes. ⌬ 9 -Tetrahydrocannabinol and endogenous cannabinoids (e.g., anandamide) initiate their agonist properties by stimulating the cannabinoid family of G protein-coupled receptors (CB 1 and CB2). The biosynthesis and physiology of anandamide is well understood, but its mechanism of uptake (resulting in signal termination by fatty acid amide hydrolase) has been elusive. Mounting evidence points to the existence of a specific anandamide transport protein; however, no direct evidence for this protein has been provided. Here, we use a potent, competitive small molecule inhibitor of anandamide uptake (LY2318912, IC 50 7.27 ؎ 0.510 nM) to identify a high-affinity, saturable anandamide transporter binding site (LY2318912; K d ‫؍‬ 7.62 ؎ 1.18 nM, Bmax ‫؍‬ 31.6 ؎ 1.80 fmol͞mg protein) that is distinct from fatty acid amide hydrolase. Systemic administration of the inhibitor into rodents elevates anandamide levels 5-fold in the brain and demonstrates efficacy in the formalin paw-licking model of persistent pain with no obvious adverse effects on motor function. Identification of the anandamide transporter binding site resolves a missing mechanistic link in endocannabinoid signaling, and in vivo results suggest that endocannabinoid transporter antagonists may provide a strategy for positive modulation of cannabinoid receptors.anandamide ͉ fatty acid amide hydrolase ͉ cannabinoid ͉ marijuana ͉ transporter E ndocannabinoids are recognized as significant intracellular lipid signaling molecules in the central nervous system with extensive control of physiological and behavioral mood and affect. Increases in endocannabinoid neurotransmission have broad therapeutic potential, including reduction of nausea and emesis (1), appetite stimulation (2), analgesia (3), anxiolytic activity (4), antispasmodic activity (5), and lowering of intraocular pressure in glaucoma (6). Identification of a specific binding site for the phytocannabinoid, ⌬ 9 -tetrahydrocannabinol (⌬ 9 -THC) (7), cloning of the cannabinoid receptors (CB 1 and CB 2 ) (8, 9), and the identification of an endogenous ligand, anandamide (N-arachidonoylethanolamide) (10), provided evidence of an endogenous cannabinoid system. Anandamide represents a class of lipid neurotransmitters that stimulate not only presynaptic and postsynaptic CB 1 receptors but also TRPV1 ion channels (11, 12), 5-hydroxytryptamine receptors (13-16), and possibly other receptors, as well as CB 2 receptors in the periphery (10,(17)(18)(19). More recently, the enzymes that are responsible for anandamide synthesis (phospholipase D) and catabolism (fatty acid amide hydrolase, FAAH) have been identified and characterized (20,21). Unlike typical neurotransmitter molecules, anandamide is synthesized in the membrane bilayer, resulting in the phospholipid precursor of anandamide, Narachidonoylphosphatidylethanolamine (22-25). Calciumacti...

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Confocal microscopy and biochemical analysis reveal spatial and functional separation between anandamide uptake and hydrolysis in human keratinocytes

Cited by 63 publications

References 60 publications

Identification of intracellular carriers for the endocannabinoid anandamide

Identification of intracellular carriers for the endocannabinoid anandamide

Aspects of endocannabinoid signaling in periimplantation biology

Identification of a high-affinity binding site involved in the transport of endocannabinoids

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