title: Dopaminergic neurons develop axonal projections to their target areas in organotypic co-cultures of the ventral mesencephalon and the striatum/prefrontal cortex.
abstract: Mesencephalic dopaminergic neurons are known to project to the prefrontal cortex (PFC) and the striatum (STR). Organotypic slice co-cultures of the ventral tegmental area/substantia nigra (VTA/SN)-complex and the PFC or STR, respectively, were used to analyze the cytoarchitectural organization of the VTA/SN-complex and the innervation pattern of the target slices by dopaminergic fibers. After 10-28 days of culturing immunocytochemistry with antibodies against tyrosine hydroxylase (TH) was performed. The VTA/SN-complex revealed in vitro an organization of TH-positive cells similar to those observed in rat brains of comparable age. TH-immunoreactive cells exhibited their typical morphology and formed long processes. No TH-immunolabeled elements were found in single cultures of PFC and STR. Tracing of VTA/SN fibers with biocytin as well as TH-immunostaining showed numerous labeled fibers in the co-cultured slices. Extensive fiber crossing was observed in the co-cultures of the VTA/SN-complex and STR but only a sparse fiber bridge in the co-cultured slices of VTA/SN-complex and PFC. The VTA/SN-complex-PFC system obviously retained several of its in vivo characteristics, e.g. the fiber network in the prefrontal cortical subareas. Our results demonstrate that TH-immunoreactive neurons develop their typical innervation pattern in slice co-cultures of VTA/SN-complex and PFC or STR, respectively. This in vitro approach may be useful for investigations of the dopaminergic function in the VTA/SN-prefrontal pathway.
authors.affiliation: Department of Pharmacology, Rudolf-Boehm Institute of Pharmacology and Toxicology, University of Leipzig, Härtelstrasse 16-18, Germany. frah@server3.medizin.uni-leipzig.de
authors.forename: Heike
authors.initials: H
authors.lastname: Franke
authors.forename: Nathalie
authors.initials: N
authors.lastname: Schelhorn
authors.forename: Peter
authors.initials: P
authors.lastname: Illes
mesh_terms: D000818:Animals; D001369:Axons / Q000502:physiology*; D002465:Cell Movement; D018920:Coculture Techniques; D004298:Dopamine / Q000502:physiology*; D007150:Immunohistochemistry; D008636:Mesencephalon / Q000166:cytology* / Q000254:growth & development; D017072:Neostriatum / Q000166:cytology* / Q000254:growth & development; D009474:Neurons / Q000502:physiology*; D009924:Organ Culture Techniques; D017397:Prefrontal Cortex / Q000166:cytology* / Q000254:growth & development; D051381:Rats; D017208:Rats, Wistar; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism
chemical_list: D014446:Tyrosine 3-Monooxygenase; D004298:Dopamine