title: Phosphodiesterase 2 inhibitors promote axonal outgrowth in organotypic slice co-cultures.
abstract: The development of appropriate models assessing the potential of substances for regeneration of neuronal circuits is of great importance. Here, we present procedures to analyze effects of substances on fiber outgrowth based on organotypic slice co-cultures of the nigrostriatal dopaminergic system in combination with biocytin tracing and tyrosine hydroxylase labeling and subsequent automated image quantification. Selected phosphodiesterase inhibitors (PDE-Is) were studied to identify their potential growth-promoting capacities. Immunohistochemical methods were used to visualize developing fibers in the border region between ventral tegmental area/substantia nigra co-cultivated with the striatum as well as the cellular expression of PDE2A and PDE10. The quantification shows a significant increase of fiber density in the border region induced by PDE2-Is (BAY60-7550; ND7001), comparable with the potential of the nerve growth factor and in contrast to PDE10-I (MP-10). Analysis of tyrosine hydroxylase-positive fibers indicated a significant increase after treatment with BAY60-7550 and nerve growth factor in relation to dimethyl sulfoxide. Additionally, a dose-dependent increase of intracellular cGMP levels in response to the applied PDE2-Is in PDE2-transfected HEK293 cells was found. In summary, our findings show that PDE2-Is are able to significantly promote axonal outgrowth in organotypic slice co-cultures, which are a suitable model to assess growth-related effects in neuro(re)generation.
authors.affiliation: Translational Centre for Regenerative Medicine TRM, University of Leipzig, Leipzig, Germany.
authors.forename: C
authors.initials: C
authors.lastname: Heine
authors.forename: K
authors.initials: K
authors.lastname: Sygnecka
authors.forename: N
authors.initials: N
authors.lastname: Scherf
authors.forename: A
authors.initials: A
authors.lastname: Berndt
authors.forename: U
authors.initials: U
authors.lastname: Egerland
authors.forename: T
authors.initials: T
authors.lastname: Hage
authors.forename: H
authors.initials: H
authors.lastname: Franke
mesh_terms: D000818:Animals; D001369:Axons / Q000187:drug effects* / Q000502:physiology; D009474:Neurons / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D009924:Organ Culture Techniques; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D013378:Substantia Nigra / Q000166:cytology / Q000187:drug effects* / Q000254:growth & development; D017557:Ventral Tegmental Area / Q000166:cytology / Q000187:drug effects* / Q000254:growth & development
chemical_list: D010726:Phosphodiesterase Inhibitors