Abstract.
The aim of the present study was to investigate whether 5,7-dihydroxytryptamine (5,7-DHT), an autofluorescent serotonin derivative, can be used as a specific marker for serotonergic or dopaminergic neurons in rat mesencephalic cultures. To this end, primary cultures were prepared from the ventral brain stem of 14-day-old Wistar rat foetuses and kept in culture for 10 days (DIV10). At DIV10, the cultures were characterized immunocytochemically with antibodies raised against tyrosine hydroxylase (TH; a marker for catecholaminergic/dopaminergic neurons) and serotonin (5-HT).
5,7-DHT labelling of the neurons was investigated after incubation with 25 µM of the serotonin derivative (plus 0.005% ascorbic acid) for 60 min at 37°C, followed by incubation with primary antibodies against TH or serotonin and a fluorescence (Cy3)-labelled secondary antibody. Using confocal laser scanning microscopy, this double immunofluorescence approach demonstrated that all cells which had accumulated 5,7-DHT additionally displayed anti-5-HT immunoreactivity, whereas no evidence was found for 5,7-DHT labelling of TH immunoreactive cells. Preincubation with the selective serotonin reuptake inhibitor fluvoxamine maleate (10 µM) prevented the loading of the 5-HT-positive cells with 5,7-DHT. In conclusion, the present data indicate that 5,7-DHT specifically labels serotonergic cells in rat midbrain cultures. Thus, 5,7-DHT can be used for the identification of living serotonergic neurons even in the presence of dopaminergic neurons.
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Franke, H., Grosche, J., Illes, P. et al. 5,7-Dihydroxytryptamine – a selective marker of dopaminergic or serotonergic neurons?. Naunyn-Schmiedeberg's Arch Pharmacol 366, 315–318 (2002). https://doi.org/10.1007/s00210-002-0621-3
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DOI: https://doi.org/10.1007/s00210-002-0621-3