Figure 3. TIGRE2.0 lines display enhanced expression in a wide range of cell types.
See also Table S2 and Figures S2–S3.
(A) Cre-dependent TIGRE2.0 reporter line design and simplified double transgenic breeding scheme. The tTA2 activity and subsequent TRE2 promoter-driven expression can be suppressed by giving doxycycline (DOX) to double transgenic mice.
(B) Representative images of native EGFP and tdTomato fluorescence in VISp of Cre;Ai139 double transgenic mice.
(C) Representative images of native EGFP and tdTomato fluorescence in VISp of Ntsr1-Cre_GN220;Ai139 mice with or without DOX.
(D) Configuration of Ai140 and representative images of native EGFP fluorescence in VISp from Cre;Ai140 mice.
(E) Configurations of GCaMP6-expressing TIGRE2.0 reporter lines.
(F) Representative images of native GCaMP6f and tdTomato fluorescence in Cre;Ai14;Ai148 mice demonstrate layer-specific expression patterns.
(G) Representative images of native GCaMP6 and tdTomato fluorescence in major inhibitory neuron classes of indicated triple or double transgenic mice.
(H) Co-localization of native GCaMP6f fluorescence and immunostaining for major neuromodulatory markers in corresponding Cre;Ai148 mice (anti-ChAT for cholinergic, anti-TH for dopaminergic, anti-NET and anti-TH for noradrenergic, and anti-TPH2 and anti-SERT for serotoninergic neurons). For region acronyms see STAR Methods.
(I) Quantification of GCaMP6 expressing cells in Cre-defined cell classes. For cortical interneuron classes (Sst, Vip and Pvalb), native GCaMP6 co-expression with tdTomato (from Ai14) was quantified within VISp and expressed as a percentage of the number of tdTomato+ cells. For neuromodulatory neuron types, native GCaMP6f co-expression with the indicated cell-type specific markers in H was expressed as a percentage of the number of marker-positive cells. Data are presented as mean ± s.e.m. (same in J); n = 3–5 mice.
(J) Quantification of GCaMP6s and tdTomato co-expression in Pvalb+ (anti-PV labeled) cells of Pvalb-IRES-Cre;Ai163 mice (n = 3–4).