title: Bioluminescent yeast assays for detecting estrogenic and androgenic activity in different matrices.
abstract: In this paper we describe the construction and use of a set of bioluminescent yeast strains for the detection of compounds that can affect androgen or estrogen receptor mediated hormonal signalling. The set includes Saccharomyces cerevisiae strains expressing human androgen receptor (AR), estrogen receptor alpha (ERalpha) or estrogen receptor beta (ERbeta), along with firefly luciferase controlled by a respective hormone responsive promoter. A constitutively luminescent strain was included in the set for determining the cytotoxicity of the sample. Yeast cells were incubated with pure chemicals or complex samples for 2.5 h, after which the signal could be detected from the cell-sample mixture after simply adding the D-luciferin substrate. The assays could be completed in one day and they required no cell lysis or centrifugation steps, which makes them suitable for high-throughput analysis of samples. Due to a short incubation time the assays are directly applicable to different sample matrices, requiring no pretreatment of the samples. The assays were used to assess the hormonal activity in moisturizing lotions as an example of a complex sample matrix known to contain endocrine disrupting chemicals. Six out of eight tested moisturisers showed high estrogenic activity, whereas no androgenic activity was observed in the samples.
authors.affiliation: Department of Biochemistry, University of Turku, FIN-20014 Turku, Finland. piiles@utu.fi
authors.forename: Piia
authors.initials: P
authors.lastname: Leskinen
authors.forename: Elisa
authors.initials: E
authors.lastname: Michelini
authors.forename: Didier
authors.initials: D
authors.lastname: Picard
authors.forename: Matti
authors.initials: M
authors.lastname: Karp
authors.forename: Marko
authors.initials: M
authors.lastname: Virta
mesh_terms: D004784:Environmental Monitoring / Q000379:methods; D004785:Environmental Pollutants / Q000032:analysis* / Q000494:pharmacology; D008163:Luminescent Measurements; D011944:Receptors, Androgen / Q000187:drug effects*; D011960:Receptors, Estrogen / Q000187:drug effects*; D015003:Yeasts / Q000502:physiology
chemical_list: D004785:Environmental Pollutants; D011944:Receptors, Androgen; D011960:Receptors, Estrogen