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

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

By: Lincoln Edwards², Paul Ernsberger²

Abstract

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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.

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Fig. 1. 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.

Affiliation(s): (2) Departments of Nutrition and Pharmacology, Case Western Reserve University School of Medicine, Cleveland, OH

Book Title: Adrenergic Receptor Protocols

Series: Methods in Molecular Biology | Volume: 126 | Pub. Date: Nov-05-1999 | Page Range: 375-390 | DOI: 10.1385/1-59259-684-3:375

Subject: Neuroscience

References

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