Transport of [ 14C]hypoxanthine by sheep choroid plexus epithelium as a monolayer in primary culture: Na +-dependent and Na +-independent uptake by the apical membrane and rapid intracellular metabolic conversion to nucleotides

Abstract

Hypoxanthine is the main product of purine metabolic degradation and previous studies have revealed that it is present in the sheep CSF and plasma in micromolar concentrations. The aim of this study was to elucidate the transport of this molecule across the sheep choroid plexus epithelium (CPE) as a monolayer in primary culture, to explore the mechanism of uptake by the apical side of the CPE and investigate the metabolic changes inside the cell. The estimated permeability of the CPE monolayer for [ 14C]hypoxanthine, [ 14C]adenine and [ 14C]guanine was low and comparable to the permeability towards the extracellular space markers. The study of [ 14C]hypoxanthine uptake by the CPE revealed two components: Na +-dependent and Na +-independent, the latter being partially mediated by the equilibrative nucleoside transporter 2. HPLC with simultaneous detection of radioactivity revealed that the majority of [ 14C]hypoxanthine inside the CPE is metabolised into [ 14C]nucleotides and [ 14C]inosine. The remaining intact [ 14C]hypoxanthine was transported across the opposite, basolateral side of CPE and appeared in the lower chamber buffer together with [ 14C]inosine. These findings indicate two possible roles of hypoxanthine uptake from the CSF by the CP epithelium in vivo: to provide material for nucleotide synthesis through the salvage pathways in the CPE, as well as to transfer excess hypoxanthine from CSF to blood.