Long-chain free fatty acids (FA) were shown to exert a regulatory function in the nucleus. However, the route of their entry remains uncertain. The aim of the present study was to examine whether the extracellular FA enter the hepatocellular nuclei. The experiments were carried out in vivo and in vitro. Intravenous administration of albumin-bound [14C]-palmitic and [14C]-linoleic acid resulted in rapid accumulation of the labels in the nuclear lipids. Unesterified [14C]-palmitic acid represented 22.4 +/- 1.7 and [14C]-linoleic acid 17.6 +/- 1.3 percent of the total lipid radioactivity. In vitro, confocal laser scanning microscopy was used to examine 12-NBD-stearate (a fluorescent derivative of stearate) translocation into the nuclei of isolated hepatocytes. It was found that 12-NBD stearate enters the nucleus and that this uptake depends on the extracellular and/or cytoplasmic concentration. It is concluded that factors (e.g. dietary) leading to alterations in the plasma FA composition and content can result in rapid changes of the nuclear FA pool and thus regulate certain nuclear processes.