A novel combined approach to detect androgenic activities with yeast based assays in Schizosaccharomyces pombe and Saccharomyces cerevisiae.
We describe the construction and validation of novel test systems for detecting androgenic activities using a combination of the yeasts Saccharomyces cerevisiae and Schizosaccharomyces pombe. By applying the reporter enhanced Green Fluorescent Protein (EGFP) the incubation time could be reduced to only 24h if compared to the classical β-galactosidase reporter (48 h). Both yeast systems were validated by analyzing the effects of seven androgens as well as five anti-androgens. One androgen (stanozolol) could be detected ten times more sensitive in S. cerevisiae than in S. pombe. Three of the five anti-androgenic substances showed no or only a slight effect in both yeast assays. The other two anti-androgens could be detected much better in S. pombe. Additionally, we could show that both yeast assays tolerated 10% urine within the media and still were capable to detect dihydrotestosterone at a concentration of 10(-8)M suggesting the use of the assays for applied doping pre-screening. In summary, the novel androgen-sensitive yeast assays have a large potential for various applications, e.g. as pre-screening in doping analysis or cattle feeding. A combination of both assays, exploiting these two phylogenetic very different yeasts, allows detection of the activity of a wide range of androgenic substances.
Wolf
Sylvi
S
Institute of Zoology, Dresden University of Technology, Germany. Sylvi.Lehmann@tu-dresden.de
Rataj
Felicitas
F
Zierau
Oliver
O
Ostermann
Kai
K
Diel
Patrick
P
Parr
Maria Kristina
MK
Vollmer
Günter
G
Rödel
Gerhard
G
D000728:Androgens / Q000032:analysis*; D001681:Biological Assay / Q000379:methods*; D005453:Fluorescence; D049452:Green Fluorescent Proteins / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000187:drug effects* / Q000235:genetics; D012568:Schizosaccharomyces / Q000187:drug effects* / Q000235:genetics
D000728:Androgens; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins