Figure 3.

Inducible and Cell Type-Specific Expression of Target Genes Controlled by FLP/CRE-Out System and Uses in Genetic and Chemogenetic Manipulation of Single-Type Neurons

(A) Confocal fluorescence images of GFP, TagBFP, and TagRFP-T before and after the treatment of heat shock; 12 h after the 2-h heat shock treatment, a single pair of ASH neurons were specifically labeled by red fluorescence. The expression patterns of sra-6p and srb-6p were ASHs/ASIs and ASHs/ADLs, respectively. Scale bar, 20 μm.

(B) Changes in Ca²⁺ transients (in curves in the left panel, and box-and-whisker in the right panel) in response to CuSO₄ solution (10 mM) in ASI neurons in worms of the indicated genotypes. No HS, without heat shock treatment; HS, with heat shock treatment. The genotypes of ZXW726 and ZXW1998 are hkdEx726[gpa-4p::R-GECO1; lin-44p::GFP] and hkdEx1998[hsp16.48p::loxP::stop1::loxP::FRT::stop2::FRT::TeTx; sra-6p::CRE::sl2::TagBFP; srb-6p::FLP::sl2::GFP; gpa-4p::R-GECO1], respectively.

(C) Changes in the Ca²⁺ signals (in curves in the left panel, and box-and-whisker in the right panel) in ASI neurons in response to CuSO₄ solution (10 mM) in worms of the indicated genotypes. Histamine of 10 mM was used to activate the HisCl1 channel. The genotype of ZXW1999 is hkdEx1999[eft-3p::loxP::stop1::loxP::FRT::stop2::FRT::HisCl1; sra-6p::CRE::sl2::TagBFP; srb-6p::FLP::sl2::GFP; gpa-4p::R-GECO1]. No His, without histamine treatment; His, with histamine treatment. Data of Ca²⁺ signals were expressed as the means ± SEM as indicated by solid traces ± gray shading and box plots with each dot representing those in each individual tested worm. The significance of statistical difference was analyzed by two-way ANOVA analysis with post hoc test of Tukey's multiple comparison correction and indicated as ∗∗∗∗p < 0.0001.