Extended Data Fig. 4: Characterization of the DREADD transgenes.

(a) Configurations of the Omp-hM4Di and the Omp-hM3Dq BAC transgene. “mem”, membrane-tagged. (b) Whole sagittal brain sections and high magnification images from the wild type, Omp-hM4Di and Omp-hM3Dq mouse, respectively (n = 3 mice for each group were examined). (c–d) Schematic diagram of the buried food test (c) and the time for wild type, Omp-hM4Di and Omp-hM3Dq mice to find the buried food (d). To test the olfaction performance, food was buried at random corner and the time that mice spent to find the food was recorded. The time for mice without clozapine injection to find buried food was recorded on day 3. On day 5, test 2 was performed to record the time for mice with clozapine injection to find the buried food (n = 13 mice for WT mice, n = 12 mice for the Omp-hM4Di transgene, n = 19 mice for the Omp-hM3Dq transgene). (e) Schematic diagram to evaluate odorant-stimulated neuronal activity of Omp-hM4Di and Omp-hM3Dq transgene with clozapine administration. Mice were tested in the cage supplied with fresh air and water underneath to maximally remove baseline odorants from the environment. (f and g) Quantification of FOS⁺ cell density in the glomerular layer (f) and ML (g) from Omp-hM4Di and wild type mice (n = 6 mice for the WT, WT clozapine and Omp-hM4Di clozapine group, n = 5 mice for the Omp-hM4Di group). (h and i) Quantification of FOS⁺ cell density in the glomerular layer (h) and ML (i) from Omp-hM3Dq and wild type mice (n = 6 mice for the WT and WT clozapine group, n = 5 mice for the Omp-hM3Dq group, n = 4 mice for the Omp-hM3Dq clozapine group). (j) Treatment scheme of the CKO_Omp-hM4Di model analysed at the pre-transforming stage. (k) Proliferation rate of mutant OPCs in the OB of CKO and CKO_Omp-hM4Di mice as treated in (j). Relative proliferation rate was calculated by the proliferation rate of mice received clozapine normalized to the average proliferation rate of mice received vehicle (n = 8 OBs for CKO with vehicle, n = 14 OBs for CKO with clozapine, n = 6 OBs for CKO_Omp-hM4Di with vehicle, n = 12 OBs for CKO_Omp-hM4Di with clozapine). (l) Treatment schedule of the CKO_Omp-hM3Dq model analysed at the pre-transforming stage. (m) Proliferation rate of mutant OPCs in the OB of CKO and CKO_Omp-hM3Dq mice as treated in (l). Relative proliferation rate was calculated by the proliferation rate of mice received clozapine normalized to the average proliferation rate of mice received vehicle (n = 6 OBs for CKO with vehicle, n = 10 OBs for CKO with clozapine, n = 6 OBs for CKO_Omp-hM3Dq with vehicle, n = 8 OBs for CKO_Omp-hM3Dq with clozapine). Of note, the same dataset of CKO and the CKO_Omp-hM3Dq model in (m) were also presented in Fig. 4k as the control for the CKO_Omp-hM3Dq_IGF1R model. Scale bars: (b), 2 mm, 200 μm. One-sided t-test in (d), (f), (g), (h), (i), (k) and (m). NS, not significance, ****p < 0.0001. Data are mean ± s.e.m.

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