Co-cultures of cerebellar slices from mice with different reelin genetic backgrounds as a model to study cortical lamination

Background: Reelin has fundamental functions in the developing and mature brain. Its absence gives rise to Reeler phenotype mice, first described cerebellar mutation. In homozygous mutants missing gene (reln-/-), neurons are incapable of correctly positioning themselves layered brain areas such as cerebral cortices. We here demonstrate that by employing ex vivo cultured slices one can reduce number animals use a non-recovery procedure analyze effects on migration Purkinje (PNs). Methods: generated mouse hybrids (L7-GFPrelnF1/) with green fluorescent protein (GFP)-tagged PNs, directly visible under fluorescence microscopy. then obtained from mice different reln genotypes demonstrated when reln-/- were co-cultured those reln+/- produced latter induced PNs partially rescue normal cortical histology. have confirmed this observation Voronoi tessellation PN dispersion. Results: images reln-/- mice, polygons larger than single-cultured same genetic background but smaller reln+/- animals. The mean roundness factor, area disorder, factor homogeneity mice cultivated singularly or co-cultivated, supporting mathematically transition clustered organization structure is supplied vivo. Conclusions: Neurobiologists primary target users 3Rs approach. They should adopt it for possibility study manipulate activity brain-secreted genetically engineered (scientific perspective), potential reduction (up 20%) used, total avoidance severe surgery (3Rs perspective).