Bitter tastant responses in the amoeba Dictyostelium correlate with rat and human taste assays

Diacylglycerol kinase (DGKA) regulates the effect of the epilepsy and bipolar disorder treatment valproic acid in Dictyostelium discoideum

“…2 C), after VPA treatment. We continued this analysis by employing a recently developed assay for monitoring acute changes in cell behaviour following drug or compound exposure, enabling quantification of cell responses ( Cocorocchio et al, 2016 , 2018 ). Here, cell behaviour was recorded using time-lapse microscopy prior to and following the addition of VPA, and computer-generated cell outlines were analysed to describe changes in cell shape (circularity) ( Fig.…”

“…As previously described ( Cocorocchio et al, 2016 , 2018 ), wild-type and transformant cells (1×10 7 ) were harvested from mid-log-phase-growing shaking cultures and washed in phosphate buffer, resuspended at 1×10 7 cells in 6 ml phosphate buffer, and pulsed with 30 nM cAMP at 6 min intervals for 5 h at 120 rpm. Cells were further washed in phosphate buffer, resuspended in 4 ml phosphate buffer and diluted 1:10.…”

“…Here, we investigate a role for DGKA in attenuating the effect of VPA in Dictyostelium . The use of Dictyostelium as a model provides the opportunity to ablate the single dgkA gene, creating a stable isogenic cell line lacking all DGK activity, and enabling the subsequent quantification of acute effects of VPA and congeners on cell behaviour and development in the absence of this enzyme ( Chang et al, 2012 ; Cocorocchio et al, 2016 , 2018 ; Xu et al, 2007 ). Previous studies show that ablation of the dgkA gene gives rise to altered development, where cells were able to form small, but relatively normal, fruiting bodies ( Egelhoff et al, 1993 ), but this study was complicated by the proposal that the enzyme functioned as a myosin II kinase, whereas it was later shown to provide DGK activity ( De La Roche et al, 2002 ; Ostroski et al, 2005 ).…”

Curcumin and derivatives function through protein phosphatase 2A and presenilin orthologues in Dictyostelium discoideum

“…It is a eukaryote, with a unique lifecycle including single-celled and multicellular stages, and contains a range of orthologues to disease-linked proteins ( Müller-Taubenberger et al, 2013 ). It has also been used to investigate the molecular actions of structurally and pharmacologically diverse compounds from bitter tastants ( Cocorocchio et al, 2015 ; Robery et al, 2011 , 2013 ), to flavonoids ( Waheed et al, 2014 ), to drugs used in the treatment of bipolar disorder ( Williams et al, 1999 , 2002 ) and epilepsy ( Chang et al, 2012 ; Xu et al, 2007 ; Elphick et al, 2012 ; Boeckeler et al, 2006 ). Several of these studies have been successfully translated to in vitro and in vivo animal models ( Chang et al, 2015 , 2016 ; Chang et al, 2013 , 2014 ).…”

“…Numerous studies using D. discoideum have employed chemical genetic approaches to identify genes controlling the cellular effects of compounds through screening mutant libraries to identify potential molecular mechanisms of compounds ( Williams et al, 1999 , 2002 ; Waheed et al, 2014 ; Robery et al, 2013 ). Several recent papers have also developed an approach to monitor the acute effects of compounds by measuring changes in cell behaviour ( Cocorocchio et al, 2015 ; Robery et al, 2011 ). D. discoideum is also widely used as a model to investigate development, where cells during starvation aggregate and differentiate to form multicellular fruiting bodies ( Marée and Hogeweg, 2001 ).…”