Coenzyme A and short-chain acyl-CoA species in control and ischemic rat brain

A rapid and reliable method was developed to quantify brain concentrations of coenzyme A (CoA) and short-chain acyl-CoAs having chain length < or = 4 carbon atoms. The method employs tissue extraction and isolation using an oligonucleotide purification cartridge and quantifies concentrations by peak area analysis following high-performance liquid chromatography (HPLC). In adult anesthetized rats subjected to 4-s high-energy microwave irradiation to stop brain metabolism, the brain concentrations of CoA, 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA), acetyl-CoA, and butyryl-CoA equaled 68.7 +/- 18.5, 2.7 +/- 1.5, 7.6 +/- 2.3, and 30.6 +/- 15.9 nmol x g(-1), respectively. After 5 min of complete ischemia, the brain concentrations of CoA and HMG-CoA increased 2- and 12-fold compared to controls, whereas acetyl-CoA and butyryl-CoA concentrations did not change. Markedly elevated levels of CoA and HMG-CoA following cerebral ischemia may reflect disturbed energy metabolism and altered formation of cholesterol and isoprenoids.