Uptake of cytidine by isolated, perfused mouse liver

Mouse livers were perfused at 22 degrees C with an oxygenated salts medium containing [5-3H]cytidine and [carboxyl-14C]inulin. The cellular uptake of cytidine was determined from the 3H content of liver samples less that present in the extracellular (inulin) space of the samples. Time courses of cytidine uptake were biphasic with initial phases which were approximately linear for 15 s and had time zero values that approximated the extracellular space. Rates of cytidine uptake derived from the initial phase of uptake evidently represented rates of membrane transport because (i) initial rates were saturable, and (ii) cytidine uptake was blocked by the nucleoside transport inhibitor, nitrobenzylthioinosine (NBMPR). Treatment of mice with the 5'-monophosphate of NBMPR (greater than 0.2 mg/kg, injected i.p.) 30-40 min prior to the perfusion also blocked cytidine entry into livers. An apparent half-saturation constant of about 10(-3) M and a maximum rate of about 1 mumol . g-1 . min-1 were estimated for the NBMPR-sensitive transport of cytidine into mouse liver cells.