Isolation, identification and synthesis of an endogenous arachidonic amide that inhibits calcium channel antagonist 1,4-dihydropyridine binding

Johnson, D. E.; Heald, Sarah L.; Dally, Robert; Janis, R.A.

doi:10.1016/0952-3278(93)90048-2

Cited by 74 publications

(30 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…For example, the inhibition of gap junctions and intracellular calcium signaling in striatal astrocytes by the non-CB 1 G-protein-coupled AEA receptor (Venance et al, 1995) or the inhibition of L-type calcium channels (Johnson et al, 1993) may prevent glutamate exocytosis and the spreading of excitotoxicity. We cannot rule out the possibility that metabolites of AEA may account for some of the observed effects.…”

Section: Discussionmentioning

confidence: 99%

Exogenous Anandamide Protects Rat Brain against Acute Neuronal InjuryIn Vivo

et al. 2001

View full text Add to dashboard Cite

The endocannabinoid anandamide [N-arachidonoylethanolamine (AEA)] is thought to function as an endogenous protective factor of the brain against acute neuronal damage. However, this has never been tested in an in vivo model of acute brain injury. Here, we show in a longitudinal pharmacological magnetic resonance imaging study that exogenously administered AEA dose-dependently reduced neuronal damage in neonatal rats injected intracerebrally with the Na ϩ /K ϩ -ATPase inhibitor ouabain. At 15 min after injury, AEA (10 mg/kg) administered 30 min before ouabain injection reduced the volume of cytotoxic edema by 43 Ϯ 15% in a manner insensitive to the cannabinoid CB 1 receptor antagonist SR141716A. At 7 d after ouabain treatment, 64 Ϯ 24% less neuronal damage was observed in AEA-treated (10 mg/kg) rats compared with control animals. Coadministration of SR141716A prevented the neuroprotective actions of AEA at this end point. In addition, (1) no increase in AEA and 2-arachidonoylglycerol levels was detected at 2, 8, or 24 hr after ouabain injection; (2) application of SR141716A alone did not increase the lesion volume at days 0 and 7; and (3) the AEA-uptake inhibitor, VDM11, did not affect the lesion volume. These data indicate that there was no endogenous endocannabinoid tone controlling the acute neuronal damage induced by ouabain. Although our data seem to question a possible role of the endogenous cannabinoid system in establishing a brain defense system in our model, AEA may be used as a structural template to develop neuroprotective agents.

show abstract

Section: Discussionmentioning

confidence: 99%

Exogenous Anandamide Protects Rat Brain against Acute Neuronal InjuryIn Vivo

et al. 2001

View full text Add to dashboard Cite

show abstract

“…It inhibited the specific binding of radiolabeled ligands to cannabinoid receptors, reduced cAMP production, and caused the inhibition of N-type calcium currents and calcium channel antagonist binding (1)(2)(3)(4)(5). Anandamide also inhibited electrically evoked contraction of vas deferens isolated from mice (1) and mimicked in vivo effects of cannabinoids such as antinociception, hypothermia, hypoactivity, and catalepsy in mice (6 -8).…”

mentioning

confidence: 98%

Partial Purification and Characterization of the Porcine Brain Enzyme Hydrolyzing and Synthesizing Anandamide

Ueda

Kurahashi

Yamamoto

et al. 1995

Journal of Biological Chemistry

304

206

View full text Add to dashboard Cite

Anandamide (arachidonylethanolamide) is known as an endogenous agonist for cannabinoid receptors. An amidohydrolase, which hydrolyzed anandamide, was solubilized from the microsomal fraction of porcine brain with 1% Triton X-100. The enzyme was partially purified by Phenyl-5PW hydrophobic chromatography to a specific activity of approximately 0.37 mol/min/mg of protein at 37°C. As assayed with 14 C-labeled substrates, the apparent K m value for anandamide was 60 M, and anandamide was more active than ethanolamides of linoleic, oleic, and palmitic acids. Ceramidase and protease activities were not detected in our enzyme preparation. The purified enzyme also synthesized anandamide from free arachidonic acid in the presence of a high concentration of ethanolamine with a specific activity of about 0.16 mol/min/mg of protein at 37°C. On the basis of cochromatographies, pH dependence, heat inactivation, and effects of inhibitors such as arachidonyl trifluoromethyl ketone, p-chloromercuribenzoic acid, diisopropyl fluorophosphate, and phenylmethylsulfonyl fluoride, it was suggested that the anandamide amidohydrolase and synthase activities were attributable to a single enzyme protein.An endogenous agonist for cannabinoid receptor was isolated from porcine brain, and this compound referred to as anandamide was identified to be arachidonylethanolamide (1). It inhibited the specific binding of radiolabeled ligands to cannabinoid receptors, reduced cAMP production, and caused the inhibition of N-type calcium currents and calcium channel antagonist binding (1-5). Anandamide also inhibited electrically evoked contraction of vas deferens isolated from mice (1) and mimicked in vivo effects of cannabinoids such as antinociception, hypothermia, hypoactivity, and catalepsy in mice (6 -8).It was shown that anandamide was rapidly degraded by an amidase (amidohydrolase) activity which was found in the membrane fraction of cultured neuroblastoma and glioma cells and homogenates of rat tissues (9). In fact, the addition of PMSF, 1 a serine protease inhibitor (9), increased an apparent affinity of anandamide for cannabinoid receptors, probably due to protecting the compound from hydrolysis (10, 11). Recently, some properties of the enzyme were reported with a microsomal preparation of rat brain (12). On the other hand, the synthesis of anandamide from free arachidonic acid and ethanolamine was shown with rat (9), bovine (13), and rabbit (14) brain and was reported to be independent of ATP and coenzyme A (14). However, the enzyme(s) hydrolyzing and synthesizing anandamide has not yet been purified and well characterized, and it is still unknown whether the two enzyme activities are attributed to a single enzyme protein or two enzymes. Enzyme Preparation-Porcine brain was obtained at a local slaughterhouse. The brain (approximately 100 g) was homogenized in 9 times the volume (v/w) of ice-cold 20 mM Tris-HCl (pH 8) containing 0.32 M sucrose with a Potter-Elvehjem homogenizer. The following procedures were performed at 4°C. The homogenate ...

show abstract

“…Metabolism of anandamide by fatty acid amide hydrolase as well as binding to intracellular proteins may also contribute to saturation kinetics in studies of anandamide uptake into cells (10)(11)(12)20). With regard to ii), to our knowledge, a carrier protein has not been isolated and characterized, but it is known that anandamide can bind to ion channels or receptors in the membrane (40)(41)(42)(43)(44). As for points iii) and iv), several inhibitors of anandamide receptors as well as of anandamide-hydrolyzing enzyme fatty acid amide hydrolase have been studied.…”

Section: Application Of the Resultsmentioning

confidence: 99%

Effect of an unstirred layer on the membrane permeability of anandamide

Bojesen

Hansen

2006

Journal of Lipid Research

View full text Add to dashboard Cite

To study the effect of an unstirred layer (UL), we have investigated the exchange efflux kinetics of anandamide at 0°C, pH 7.3, from albumin-free as well as from albumin-filled human red blood cell ghosts to media of various BSA concentrations ([BSA] o ). The rate constant (k m ) of unidirectional flux from the outer membrane leaflet to BSA in the medium increased with the square root of [BSA] o in accordance with the existence of a UL, which is a water layer adjacent to the membrane that is not subject to the same gross mixing that takes place in the rest of the medium. From k m , it is possible to calculate the rate constant of anandamide dissociation from BSA (k 1 ) if we know the membrane binding of anandamide, the equilibrium dissociation constant of BSA-anandamide complexes, and the diffusion constant of anandamide. We estimated k 1 to be 3.33 6 0.27 s 21. The net flux of [ 3 H]anandamide is balanced by an equal and opposite movement of nonradioactive anandamide in exchange efflux experiments. This means that our results are also valid for uptake. We show that for anandamide with rapid membrane translocation, UL causes a significant resistance to cellular uptake. Depicting the rate of anandamide uptake as a function of equilibrium water phase concentrations results in a parabolic uptake dependence. Such apparent "saturation kinetics" is often interpreted as indicating the involvement of transport proteins. The validity of such an interpretation is discussed.-Bojesen, I. N., and H. S. Hansen. Effect of an unstirred layer on the membrane permeability of anandamide. J. Lipid Res. 2006. 47: 561-570. Supplementary key words red blood cell membranes . erythrocyte ghosts . membrane binding . exchange efflux . rate constant of anandamide dissociation from albumin . diffusion coefficient of anandamide

show abstract

Isolation, identification and synthesis of an endogenous arachidonic amide that inhibits calcium channel antagonist 1,4-dihydropyridine binding

Cited by 74 publications

References 15 publications

Exogenous Anandamide Protects Rat Brain against Acute Neuronal InjuryIn Vivo

Exogenous Anandamide Protects Rat Brain against Acute Neuronal InjuryIn Vivo

Partial Purification and Characterization of the Porcine Brain Enzyme Hydrolyzing and Synthesizing Anandamide

Effect of an unstirred layer on the membrane permeability of anandamide

Contact Info

Product

Resources

About