title: Systematic evaluation of buffer influences on the development of Dictyostelium discoideum.
abstract: Development and cell differentiation are key features of the social amoeba Dictyostelium discoideum. Already at early developmental stages, the gene expression profile changes in the amoebae to make the cells aggregation competent. In the laboratory, development starts when the cells are washed free of nutrients. For this purpose, various non-nutrient buffers are used in different laboratories. However, to date, it is not clear if different buffers have different influences on the development of D. discoideum. Therefore, we investigated systematically the influence of six widely used buffers on the development of D. discoideum. Investigation was done at the phenotypical, biochemical, and molecular level. The results show that some of the investigated buffers show clear differences in the phenotypical outcome of the developmental cycle, at a biochemical level as measured in the response to cAMP, and/or at a molecular level as measured in the expression of early developmental marker genes. According to our results buffer compositions should be considered carefully for all developmental experiments with D. discoideum, especially when gene expression will be investigated.
authors.affiliation: Institute for Biology-Microbiology, Department of Biology, Chemistry, Pharmacy, Freie Universität Berlin, Königin-Luise-Str. 12-16, 14195, Berlin, Germany.
authors.forename: Johanna
authors.initials: J
authors.lastname: Márquez López
authors.affiliation: Institute for Biology-Microbiology, Department of Biology, Chemistry, Pharmacy, Freie Universität Berlin, Königin-Luise-Str. 12-16, 14195, Berlin, Germany.
authors.forename: Anja
authors.initials: A
authors.lastname: Sulzmann
authors.affiliation: Institute for Biology-Microbiology, Department of Biology, Chemistry, Pharmacy, Freie Universität Berlin, Königin-Luise-Str. 12-16, 14195, Berlin, Germany. sascha.thewes@fu-berlin.de.
authors.forename: Sascha
authors.identifier: http://orcid.org/0000-0001-5487-3286
authors.initials: S
authors.lastname: Thewes
mesh_terms: D001419:Bacteria; D003470:Culture Media / Q000737:chemistry; D004023:Dictyostelium / Q000235:genetics* / Q000254:growth & development* / Q000528:radiation effects; D008027:Light; D010641:Phenotype; D010710:Phosphates / Q000737:chemistry; D017680:Potassium Compounds / Q000737:chemistry; D059467:Transcriptome
chemical_list: D003470:Culture Media; D010710:Phosphates; D017680:Potassium Compounds; C013216:potassium phosphate
keywords: Buffer influence; Development; Dictyostelium; Gene expression; cAMP