anandamide and Astrocytoma

anandamide has been researched along with Astrocytoma* in 5 studies

Other Studies

5 other study(ies) available for anandamide and Astrocytoma

ArticleYear
Evidence against the presence of an anandamide transporter.
    Proceedings of the National Academy of Sciences of the United States of America, 2003, Apr-01, Volume: 100, Issue:7

    On the basis of temperature dependency, saturability, selective inhibition, and substrate specificity, it has been proposed that an anandamide transporter exists. However, all of these studies have examined anandamide accumulation at long time points when downstream effects such as metabolism and intracellular sequestration are operative. In the current study, we have investigated the initial rates (<1 min) of anandamide accumulation in neuroblastoma and astrocytoma cells in culture and have determined that uptake is not saturable with increasing concentrations of anandamide. However, anandamide hydrolysis, after uptake in neuroblastoma cells, was saturable at steady-state time points (5 min), suggesting that fatty acid amide hydrolase (FAAH) may be responsible for observed saturation of uptake at long time points. In general, arvanil, olvanil, and N-(4-hydroxyphenyl)arachidonylamide (AM404) have been characterized as transport inhibitors in studies using long incubations. However, we found these "transport inhibitors" did not inhibit anandamide uptake in neuroblastoma and astrocytoma cells at short time points (40 sec or less). Furthermore, we confirmed that these inhibitors in vitro were actually inhibitors of FAAH. Therefore, the likely mechanism by which the transport inhibitors raise anandamide levels to exert pharmacological effects is by inhibiting FAAH, and they should be reevaluated in this context. Immunofluorescence has indicated that FAAH staining resides mainly on intracellular membranes of neuroblastoma cells, and this finding is consistent with our observed kinetics of anandamide hydrolysis. In summary, these data suggest that anandamide uptake is a process of simple diffusion. This process is driven by metabolism and other downstream events, rather than by a specific membrane-associated anandamide carrier.

    Topics: Arachidonic Acids; Astrocytoma; Biological Transport; Cannabinoids; Capsaicin; Carrier Proteins; Endocannabinoids; Humans; Immunohistochemistry; Kinetics; Neuroblastoma; Polyunsaturated Alkamides; Tumor Cells, Cultured

2003
The CB1 cannabinoid receptor is coupled to the activation of protein kinase B/Akt.
    The Biochemical journal, 2000, Apr-15, Volume: 347, Issue:Pt 2

    Cannabinoids exert most of their effects in the central nervous system through the CB(1) cannabinoid receptor. This G-protein-coupled receptor has been shown to be functionally coupled to inhibition of adenylate cyclase, modulation of ion channels and activation of extracellular-signal-regulated kinase. Using Chinese hamster ovary cells stably transfected with the CB(1) receptor cDNA we show here that Delta(9)-tetrahydrocannabinol (THC), the major active component of marijuana, induces the activation of protein kinase B/Akt (PKB). This effect of THC was also exerted by the endogenous cannabinoid anandamide and the synthetic cannabinoids CP-55940 and HU-210, and was prevented by the selective CB(1) antagonist SR141716. Pertussis toxin and wortmannin blocked the CB(1) receptor-evoked activation of PKB, pointing to the sequential involvement of a G(i)/G(o) protein and phosphoinositide 3'-kinase. The functionality of the cannabinoid-induced stimulation of PKB was proved by the increased phosphorylation of glycogen synthase kinase-3 serine 21 observed in cannabinoid-treated cells and its prevention by SR141716 and wortmannin. Cannabinoids activated PKB in the human astrocytoma cell line U373 MG, which expresses the CB(1) receptor, but not in the human promyelocytic cell line HL-60, which expresses the CB(2) receptor. Data indicate that activation of PKB may be responsible for some of the effects of cannabinoids in cells expressing the CB(1) receptor.

    Topics: Adenylate Cyclase Toxin; Androstadienes; Arachidonic Acids; Astrocytoma; Calcium-Calmodulin-Dependent Protein Kinases; Cannabinoids; Dronabinol; Endocannabinoids; Enzyme Activation; Glycogen Synthase Kinase 3; Glycogen Synthase Kinases; GTP-Binding Protein alpha Subunits, Gi-Go; Heterotrimeric GTP-Binding Proteins; HL-60 Cells; Humans; Pertussis Toxin; Phosphatidylinositol 3-Kinases; Phosphoinositide-3 Kinase Inhibitors; Phosphorylation; Polyunsaturated Alkamides; Protein Serine-Threonine Kinases; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-akt; Receptors, Cannabinoid; Receptors, Drug; Tumor Cells, Cultured; Virulence Factors, Bordetella; Wortmannin

2000
Carrier-mediated transport and enzymatic hydrolysis of the endogenous cannabinoid 2-arachidonylglycerol.
    Neuroreport, 2000, Apr-27, Volume: 11, Issue:6

    The human astrocytoma cell line CCF-STTGI accumulates [3H]2-AG through an Na(+)- and energy-independent process, with a Km of 0.7 +/- 0.1 microM. Non-radioactive 2-AG, anandamide or the anandamide transport inhibitor 4-hydroxyphenyl arachidonamide inhibit [3H]2-AG uptake with half-maximal inhibitory concentrations (IC50) of 5.5 +/- 1.0 microM, 4.2 +/- 0.3 microM and 1.8 = 0.1 microM, respectively. A variety of lipid transport substrates and inhibitors interfere with neither [3H]2-AG nor [3H]anandamide uptake. These results suggest that 2-AG and anandamide are internalized in astrocytoma cells through a common carrier-mediated mechanism. After incubation with [3H]2-AG, radioactivity is recovered in phospholipids, monoacylglycerols (unmetabolized [3H]2-AG), free fatty acids ([3H]arachidonate) and, to a minor extent, diacylglycerols and triacylglycerols. Arachidonic acid (100 microM) and triacsin C (10 microM), an acyl-CoA synthetase inhibitor, prevent incorporation of [3H]arachidonic acid in phospholipids and significantly reduce [3H]2-AG transport. Thus, the driving force for 2-AG internalization may derive from the hydrolysis of 2-AG to arachidonate and the subsequent incorporation of this fatty acid into phospholipids.

    Topics: Arachidonic Acid; Arachidonic Acids; Astrocytoma; Binding, Competitive; Biological Transport; Calcium Channel Blockers; Carrier Proteins; Chromatography, Thin Layer; Dose-Response Relationship, Drug; Endocannabinoids; Enzyme Inhibitors; Glycerides; Glycerol; Humans; Hydrolysis; Intracellular Fluid; Lipid Metabolism; Lipids; Neurotransmitter Agents; Phospholipids; Polyunsaturated Alkamides; Triazenes; Tritium; Tumor Cells, Cultured

2000
Anandamide transport inhibition by the vanilloid agonist olvanil.
    European journal of pharmacology, 1999, Jan-01, Volume: 364, Issue:1

    The structural similarities between the anandamide transport inhibitor N-(4-hydroxyphenyl)-arachidonylamide (AM404) and the synthetic vanilloid agonist olvanil [(N-vanillyl)-9-oleamide], prompted us to investigate the possibility that olvanil may interfere with anandamide transport. The intracellular accumulation of [3H]anandamide by human astrocytoma cells was prevented by olvanil with a Ki value of 14.1+/-7.1 microM. By contrast, capsaicin [(8-methyl-N-vanillyl)-6-noneamide], a plant-derived vanilloid agonist, and capsazepine (N-[2-(4-chlorophenyl)ethyl]-1,3,4,5-tetrahydro-7,8-dihydroxy-2 H-2-benzazepine-2-carbothioamide), a vanilloid antagonist, had no such effect (Ki > 100 microM). These results indicate that, although less potent than AM404 (Ki 2.1+/-0.2 microM), olvanil may reduce anandamide clearance at concentrations similar to those needed for vanilloid receptor activation.

    Topics: Amidohydrolases; Animals; Anti-Inflammatory Agents, Non-Steroidal; Arachidonic Acids; Astrocytoma; Biological Transport; Brain; Capsaicin; Depression, Chemical; Endocannabinoids; Humans; Polyunsaturated Alkamides; Rats; Receptors, Drug; Tumor Cells, Cultured

1999
Structural determinants for recognition and translocation by the anandamide transporter.
    Proceedings of the National Academy of Sciences of the United States of America, 1999, May-11, Volume: 96, Issue:10

    The biological actions of anandamide (arachidonylethanolamide), an endogenous cannabinoid lipid, are terminated by a two-step inactivation process consisting of carrier-mediated uptake and intracellular hydrolysis. Anandamide uptake in neurons and astrocytes is mediated by a high-affinity, Na+-independent transporter that is selectively inhibited by N-(4-hydroxyphenyl)-arachidonamide (AM404). In the present study, we examined the structural determinants governing recognition and translocation of substrates by the anandamide transporter constitutively expressed in a human astrocytoma cell line. Competition experiments with a select group of analogs suggest that substrate recognition by the transporter is favored by a polar nonionizable head group of defined stereochemical configuration containing a hydroxyl moiety at its distal end. The secondary carboxamide group interacts favorably with the transporter, but may be replaced with either a tertiary amide or an ester, suggesting that it may serve as hydrogen acceptor. Thus, 2-arachidonylglycerol, a putative endogenous cannabinoid ester, also may serve as a substrate for the transporter. Substrate recognition requires the presence of at least one cis double bond situated at the middle of the fatty acid carbon chain, indicating a preference for ligands whose hydrophobic tail can adopt a bent U-shaped conformation. On the other hand, uptake experiments with radioactively labeled substrates show that no fewer than four cis nonconjugated double bonds are required for optimal translocation across the cell membrane, suggesting that substrates are transported in a folded hairpin conformation. These results outline the general structural requisites for anandamide transport and may assist in the development of selective inhibitors with potential clinical applications.

    Topics: Arachidonic Acids; Astrocytoma; Binding, Competitive; Biological Transport; Carrier Proteins; Cell Line; Endocannabinoids; Ethanolamines; Glycerides; Humans; Kinetics; Models, Molecular; Molecular Conformation; Molecular Structure; Polyunsaturated Alkamides; Substrate Specificity

1999