Quantitative brain autoradiography of [9, 10‐3H]palmitic acid incorporation into brain lipids

Abstract

The distribution of radioactivity within brain metabolic compartments was examined following the intravenous injection of [9,10‐³H]palmitate into awake rats. Brain radioactivity reached a maximum value by 15 min after [9,10‐³H]palmitate injection and remained unchanged for at least 4 hr. Regional differences in radioactivity could be determined with high resolution by quantitative autoradiography, at the level of cell layers within the hippocampus and cerebral cortex, and between striosomes of the caudate nucleus. Regional brain radioactivities were converted to normalized regional radioactivities (k) by dividing them by the integrated plasma fatty acid radioactivity (integrated over the time course of the experiment). These values reflected incorporation mainly into brain phospholipids; radioactivity due to nonlipid components was minimal. Indeed, about 85% of brain radioactivity was within lipids between 5 min and 4 hr postinjection, the remainder being equally divided between protein‐associated pellet and aqueous‐soluble metabolites. The major lipids labeled were phospholipids, particularly phosphatidylcholine, which contained about 75% of phospholipid radioactivity. The results show that [9,10‐³H]palmitate can be used to examine incorporation of plasma palmitate into individual brain regions via quantitative autoradiography. Furthermore, the tracer is a rather selective marker for phosphatidylcholine and can be used to examine turnover and synthesis of this phospholipid. [9,10‐³H]palmitate has advantages over [U‐¹⁴C]palmitate for autoradiographic studies of incorporation; following the ¹⁴C‐tracer, significant label even at 4 hr after injection is in nonlipid compartments (glutamate and aspartate), and the long path length of ¹⁴C limits resolution at the cell layer level.