The uptake of L-lactate was investigated in neuronal primary cultures derived from embryonic rat brain with a radioactive tracer method. After preincubation of the cells in glucose-free buffer for 30 min, uptake increased with time for at least 10 min. A saturable component of uptake was found with half-maximal uptake at 10 mM lactate. This saturable component was abolished in the presence of 10 mM alpha-cyano-4-hydroxcinnamic acid. In addition, a non-saturable component dominated the uptake at high concentrations of lactate. Uptake was accelerated with decreasing pH, and was inhibited considerably by pyruvate. It is concluded that neurons are endowed with a lactate transport system which resembles in its properties the monocarboxylate carrier of peripheral tissues.