Release of choline in the isolated heart, an indicator of ischemic phospholipid degradation and its protection by ischemic preconditioning: No evidence for a role of phospholipase D Annette Bru ¨hl a , Gerd Hafner b,1 , Konrad Lo ¨ffelholz a, * a Department of Pharmacology, Johannes-Gutenberg-University of Mainz, Obere Zahlbacher Str. 67, D-55101 Mainz, Germany b Department of Laboratory Medicine, Johannes-Gutenberg-University of Mainz, Obere Zahlbacher Str. 67, D-55101 Mainz, Germany Received 2 December 2003; accepted 17 March 2004 Abstract The release of choline as a water-soluble product of phospholipid hydrolysis was measured in the perfusate of rat hearts to monitor ischemic membrane degradation and its protection by ischemic preconditioning (IPC). Hearts were subjected to global ischemia (GI; 30 min of no-flow) followed by 60 min of reperfusion. To induce IPC, GI was preceded by four no-flow episodes of 5 min each. Deleterious consequences of GI and reperfusion, namely coronary flow reduction, incidence of arrhythmias and release of cardiac troponin T, were significantly attenuated by IPC. The release of choline increased during reperfusion in a biphasic manner: a first phase peaked immediately after GI and was followed by a second, delayed phase indicating choline release caused during reperfusion. Only the second phase was blocked by both IPC and by AACOCF 3 (5 AM), an inhibitor of cytosolic phospholipase A 2 . The activity of phospholipase D (PLD) was unchanged after GI or IPC or GI plus IPC. In conclusion, choline release into heart perfusate was found to be a useful real-time indicator of phospholipid degradation caused by GI and by reperfusion and its protection by IPC. The results supplement previous observations on the accumulation of fatty acids in the phospholipid pool. There was no evidence for PLD activation by GI or IPC. D 2004 Published by Elsevier Inc. Keywords: Ischemia; Preconditioning; Heart; Choline; Phospholipids; Phospholipases 0024-3205/$ - see front matter D 2004 Published by Elsevier Inc. doi:10.1016/j.lfs.2004.03.019 * Corresponding author. Tel.: +49-6131-3933260; fax: +49-6131-3936611. E-mail address: loeffelh@uni-mainz.de (K. Lo ¨ffelholz). 1 Present address: Zentrum fu ¨r Labormedizin und Mikrobiologie, Herwarthstr. 100, 45138 Essen, Germany. www.elsevier.com/locate/lifescie Life Sciences 75 (2004) 1609 – 1620