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Assay of Arachidonic Acid Release Coupled to α1- and α2-Adrenergic Receptors

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Adrenergic Receptor Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 126))

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Abstract

AA is the precursor for prostaglandins, eicosanoids, and the endogenous cannabinoid ligands, such as anandamide (1). The major pathways that lead to the liberation of free AA from phospholipids are shown in Fig. 1. Phospholipase A2 (PLA2) provides a direct pathway for AA release and is the most common source of cellular AA liberation. PLA2 is the primary source of free AA in most cell systems, including neurons. Physiological regulation of PLA2 occurs primarily through the action of calcium, which binds to several PLA2 isoforms and is required for activity (2,3). The low-molecular-weight secretory PLA2 isoforms require millimolar calcium, whereas the cytosolic classes of PLA2 need only micromolar concentrations. Calcium is required for translocation to membranes, where the cytosolic form of the enzyme can have access to its phospholipid substrate. Other isoforms are completely independent of calcium, but these variants may be primarily involved in membrane remodeling rather than signal transduction (4). The calcium-independent forms of PLA2 play an important role in the rapid incorporation of free AA into phospholipids, which is taken advantage of in the AA release assay described here.

Possible pathways for generation of AA from phospholipids. Hypothetical model of the biochemical pathways contributing to the liberation of AA. AA = arachidonic acid, PLA2=phospholipase A2, PLC=phospholipase C, PLD=phospholipase D, PA=phosphatidic acid, DAG=diacylglyceride, MAG=monoacylglyceride, EtOH=ethanol, PEt=phosphatidylethanol.

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Authors and Affiliations

  1. Departments of Nutrition and Pharmacology, Case Western Reserve University School of Medicine, Cleveland, OH

    Lincoln Edwards & Paul Ernsberger

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  1. Lincoln Edwards
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  2. Paul Ernsberger
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Editors and Affiliations

  1. Oregon Regional Primate Research Center, Oregon Health Sciences University, Beaverton, OR

    Curtis A. Machida

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© 2000 Humana Press Inc., Totowa, NJ

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Edwards, L., Ernsberger, P. (2000). Assay of Arachidonic Acid Release Coupled to α1- and α2-Adrenergic Receptors. In: Machida, C.A. (eds) Adrenergic Receptor Protocols. Methods in Molecular Biology™, vol 126. Humana Press. https://doi.org/10.1385/1-59259-684-3:375

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  • DOI: https://doi.org/10.1385/1-59259-684-3:375

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-602-4

  • Online ISBN: 978-1-59259-684-3

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