Rat mesencephalic neuronal cells cultured for different periods as a model of dopamine transporter ontogenesis

Ventral mesencephalic neurons contained only low-affinity and sodium-independent binding sites of [3H]WIN 35,428 (marker of dopamine transporter) during the first 10 d in primary cultures. These sites were present in cytosol, and they are not very probably related to dopamine transporter. After 12 d in culture, membrane-bound, high-affinity, and sodium-dependent [3H]WIN 35,428 binding sites were detected. In membranes prepared from cells 14 d in culture, cocaine displaced [3H]WIN 35,428 binding with similar potency to that in striatal membranes of adult rat brain. The high-affinity [3H]WIN 35,428 binding sites in mesencephalic neuronal cell cultures are very probably related to dopamine transporter. The development of high-affinity [3H]WIN 35,428 binding sites in neurons cultured for different time periods could be a useful model of dopamine transporter ontogenesis.