Article Figures & Data
Figures
- Figure 1.
Cell-type-specific expression of GCaMP3 in OBs and distinct OSN axon trajectories in IPA-responsive glomeruli. A1–A4, Confocal images of OBs in Cre-dependent GCaMP3 reporter mice crossed with OMP-Cre (A1), Gad2-Cre (A2), DAT-Cre (A3), and Pcdh21-Cre (A4) mice. Magnified views of the dashed squares are shown in the insets in A2, A3. GL, EPL, MCL, and GCL indicate GL, external plexiform layer, MCL, and GCL, respectively. B, Process for the DiI labeling of OSN axons. B1, Resting fluorescence of GCaMP3 in the dorsal OB of OMP-Cre mouse. B2, IPA (0.02%)-responsive homologous glomeruli were observed by calcium imaging [color scale indicates ΔF/F0 (%) of GCaMP3 signal]. B3, Brightfield image after DiI implantation. B4, DiI fluorescence 30 min after DiI implantation. B5, DiI fluorescence 8 h after DiI implantation. The locations of IPA-responsive glomeruli are indicated by the white dotted circles. C, Two-photon microscopy image of DiI-labeled glomeruli and OSN axons 2 d after DiI implantation. D, Magnified image of area denoted by orange dotted square in C associated with lateral glomerulus. E, Magnified image of area denoted by yellow dotted square in C associated with medial glomerulus. Two-photon microscopy images of OSN axons that transverse the lateral and medial surface of OB; orange and yellow dotted ellipses (in C–E) represent major axonal projections from lateral and medial glomeruli. F, Two-photon microscopy images of areas of lateral/medial border denoted by blue dotted square in C. Red asterisks indicate axon termination in several glomeruli in which the labeled OSNs probably passed through the surface of the DiI-implanted glomeruli. Some minor axons which did not show clear axon terminations in a glomerulus were observed in area 2 in this case. Ant., anterior. Lat., lateral. Scale bars: 100 μm (A), 200 μm (B, C), 50 μm (D, E), and 100 μm (F).
- Figure 2.
Odor-evoked response maps and traces among different types of OB neurons. Odor-evoked response maps (A) and traces (B) from OMP-Cre, Gad2-Cre, DAT-Cre, and Pcdh21-Cre mice. Resting GCaMP3 fluorescence images are displayed in the left panels in A. Pseudocolored images in the middle and right panels in A indicate responses to 0.02% IPA and PEA, respectively. Yellow dotted lines and circles in each image show approximately the edge of the left OB and homologous glomeruli evoked by 0.02% IPA and PEA, respectively. The color scales represent ΔF/F0 (%) of GCaMP3 signal. Traces shown in B represent GCaMP3 fluorescence changes from two subsets of homologous glomeruli evoked by indicated concentrations of IPA and PEA shown in A. Red and blue traces indicate lateral and medial glomeruli, respectively. Gray bars under each trace and vertical dotted lines indicate the timing of odor stimulation. Respiration signals are also shown under the traces.
- Figure 3.
Onset latencies of medial and lateral glomeruli. A, Scatter plots displaying the distributions of onset latencies of medial and lateral glomeruli, which responded to PEA and IPA stimuli. x and y axes indicate onset latencies of medial and lateral glomerular responses, respectively. Individual green and orange dots indicate single trial data of 0.02% and 0.002% PEA or IPA. Gray lines indicate the equal onset latency time points of medial and lateral glomerular responses. B, Box plots displaying the distributions of the differences of onset latencies between medial and lateral glomeruli responses to 0.02% and 0.002% PEA or IPA. Red horizontal lines in each box indicate the medians. Quartiles are shown as whiskers. NS, not significant (two-tailed paired Student’s t test).
- Figure 4.
Rise times of medial and lateral glomeruli. A, Scatter plots displaying the distributions of rise times of the medial and lateral glomeruli, which responded to PEA and IPA stimuli. x and y axes indicate rise times of medial and lateral glomerular responses, respectively. Individual green and orange dots indicate single trial data of 0.02% and 0.002% PEA or IPA. Gray lines indicate the equal rise time points of medial and lateral glomerular responses. B, Box plots displaying the distributions of the differences of rise times between medial and lateral glomerular responses to 0.02% and 0.002% PEA or IPA. Red horizontal lines in each box indicate the medians. Quartiles are shown as whiskers. NS, not significant (two-tailed paired Student’s t test).
- Figure 5.
Decay times of medial and lateral glomeruli. A, Scatter plots displaying the distributions of decay times of the medial and lateral glomeruli, which responded to PEA and IPA stimuli. x and y axes indicate decay times of medial and lateral glomerular responses, respectively. Individual green and orange dots indicate single trial data of 0.02% and 0.002% PEA or IPA. Gray lines indicate the equal decay time points of medial and lateral glomerular responses. B, Box plots displaying the distributions of the differences of decay times between medial and lateral glomerular responses to 0.02% and 0.002% PEA or IPA. Red horizontal lines in each box indicate the medians. Quartiles are shown as whiskers. NS, not significant (two-tailed paired Student’s t test).
- Figure 6.
Peak amplitude of medial and lateral glomerular responses. A, Scatter plots displaying the distributions of peak amplitudes of medial and lateral glomerular responses to PEA and IPA stimuli. x- and y-axes indicate peak amplitudes of medial and lateral glomerular responses, respectively. Individual green and orange dots indicate single trial data of 0.02% and 0.002% PEA or IPA. Gray lines indicate the equal peak amplitudes of medial and lateral glomerular responses. B, Box plots displaying the distributions of the differences of peak amplitudes of medial and lateral glomerular responses to 0.02% and 0.002% PEA or IPA. Red horizontal lines in each box indicate the medians. Quartiles are shown as whiskers. NS, not significant (two-tailed paired Student’s t test).
- Figure 7.
Respiration-locked calcium fluctuations of medial and lateral glomeruli. A, Schematic illustration of data collection time points and power spectral analysis. The prestimulation and odor stimulation periods are defined in left panel. Middle and right panels are representative examples of power spectral analyses with large frequency ranges in single trials. Respiration-locked fluctuations of calcium signals were prominently observed at 2–4 Hz (arrows). B, Representative results of power spectral analyses of calcium signals during prestimulation (upper rows) and odor stimulation (lower rows) periods in neurons from GCaMP-expressing OMP-Cre, Gad2-Cre, DAT-Cre, and Pcdh21-Cre mice. The calcium response traces of these spectra are shown as 0.02% IPA response signals in Fig. 2B. Blue and red traces indicate medial and lateral glomeruli, respectively (see also Extended Data Fig. 7-1). C, Comparisons of power spectra between the homologous glomeruli during prestimulation (upper rows) and odor stimulation (lower rows) period in neurons from OMP-Cre, Gad2-Cre, DAT-Cre, and Pcdh21-Cre mice. The ratios of peak powers were calculated by dividing the medial glomerular power by the lateral glomerular power in each trial. The powers were calculated after correction for the white-noise background (see Materials and Methods). D, Comparisons of peak power between the prestimulation and odor stimulation periods of the same glomeruli in each cell type. The power ratios were calculated by dividing the power value in the odor stimulation period by that in the prestimulation period after the subtraction of background level (see text); *p < 0.05, **p < 0.01, ***p < 0.001 two-tailed paired Student’s t test.
Tables
Figure Mice Odorant Conc, % N Mean
(medial), sSD
(medial), sMean
(lateral), sSD
(lateral), s3A OMP PEA 0.02 279 0.138 0.086 0.142 0.086 3A OMP PEA 0.002 194 0.153 0.096 0.153 0.098 3A OMP IPA 0.02 297 0.151 0.101 0.154 0.100 3A OMP IPA 0.002 185 0.155 0.095 0.158 0.097 3A Gad2 PEA 0.02 106 0.103 0.099 0.108 0.096 3A Gad2 PEA 0.002 84 0.125 0.083 0.128 0.081 3A Gad2 IPA 0.02 119 0.112 0.110 0.113 0.104 3A Gad2 IPA 0.002 62 0.143 0.105 0.146 0.101 3A DAT PEA 0.02 216 0.157 0.132 0.158 0.127 3A DAT PEA 0.002 118 0.162 0.128 0.162 0.125 3A DAT IPA 0.02 184 0.148 0.096 0.152 0.098 3A DAT IPA 0.002 123 0.160 0.098 0.166 0.093 3A Pcdh21 PEA 0.02 199 0.183 0.125 0.185 0.129 3A Pcdh21 PEA 0.002 130 0.197 0.126 0.202 0.126 3A Pcdh21 IPA 0.02 241 0.179 0.128 0.178 0.123 3A Pcdh21 IPA 0.002 176 0.210 0.147 0.214 0.144 4A OMP PEA 0.02 279 0.638 0.305 0.631 0.300 4A OMP PEA 0.002 194 0.732 0.307 0.724 0.307 4A OMP IPA 0.02 297 0.658 0.300 0.653 0.304 4A OMP IPA 0.002 185 0.679 0.279 0.674 0.275 4A Gad2 PEA 0.02 106 0.425 0.302 0.441 0.300 4A Gad2 PEA 0.002 84 0.489 0.290 0.490 0.297 4A Gad2 IPA 0.02 119 0.486 0.301 0.496 0.314 4A Gad2 IPA 0.002 62 0.549 0.377 0.543 0.381 4A DAT PEA 0.02 216 0.651 0.357 0.656 0.352 4A DAT PEA 0.002 118 0.656 0.324 0.654 0.328 4A DAT IPA 0.02 184 0.594 0.310 0.606 0.308 4A DAT IPA 0.002 123 0.597 0.311 0.602 0.301 4A Pcdh21 PEA 0.02 199 0.590 0.327 0.580 0.321 4A Pcdh21 PEA 0.002 130 0.521 0.326 0.518 0.330 4A Pcdh21 IPA 0.02 241 0.590 0.313 0.585 0.327 4A Pcdh21 IPA 0.002 176 0.586 0.332 0.579 0.330 5A OMP PEA 0.02 279 1.007 0.356 1.003 0.358 5A OMP PEA 0.002 194 0.883 0.297 0.879 0.301 5A OMP IPA 0.02 297 0.939 0.306 0.934 0.313 5A OMP IPA 0.002 185 0.945 0.335 0.940 0.327 5A Gad2 PEA 0.02 106 0.890 0.340 0.888 0.337 5A Gad2 PEA 0.002 84 0.858 0.346 0.857 0.349 5A Gad2 IPA 0.02 119 0.917 0.365 0.909 0.358 5A Gad2 IPA 0.002 62 0.846 0.349 0.845 0.352 5A DAT PEA 0.02 216 0.936 0.386 0.930 0.388 5A DAT PEA 0.002 118 0.850 0.316 0.853 0.311 5A DAT IPA 0.02 184 0.926 0.356 0.928 0.361 5A DAT IPA 0.002 123 0.839 0.295 0.849 0.301 5A Pcdh21 PEA 0.02 199 1.101 0.412 1.098 0.400 5A Pcdh21 PEA 0.002 130 0.975 0.416 0.969 0.393 5A Pcdh21 IPA 0.02 241 1.083 0.399 1.088 0.396 5A Pcdh21 IPA 0.002 176 1.041 0.407 1.032 0.392 6A OMP PEA 0.02 279 3.114 1.635 3.099 1.597 6A OMP PEA 0.002 194 2.244 0.963 2.248 0.979 6A OMP IPA 0.02 297 3.096 1.741 3.123 1.772 6A OMP IPA 0.002 185 2.469 1.078 2.478 1.077 6A Gad2 PEA 0.02 106 5.910 2.435 5.869 2.360 6A Gad2 PEA 0.002 84 4.309 1.712 4.349 1.831 6A Gad2 IPA 0.02 119 6.442 3.217 6.294 3.029 6A Gad2 IPA 0.002 62 4.695 1.982 4.626 1.950 6A DAT PEA 0.02 216 5.895 2.866 5.870 2.763 6A DAT PEA 0.002 118 4.739 2.480 4.693 2.312 6A DAT IPA 0.02 184 5.851 2.892 5.877 2.991 6A DAT IPA 0.002 123 4.266 2.174 4.290 2.142 6A Pcdh21 PEA 0.02 199 7.565 4.335 7.531 4.281 6A Pcdh21 PEA 0.002 130 5.506 2.356 5.601 2.302 6A Pcdh21 IPA 0.02 241 7.610 4.344 7.576 4.301 6A Pcdh21 IPA 0.002 176 5.532 2.858 5.444 2.744 Figure Mice Odorant Conc, % N Lower 95% CI, ms Upper 95% CI, ms Mean, ms SD, ms p 3B OMP PEA 0.02 279 –9.6 1.2 −4.2 45.9 0.130 3B OMP PEA 0.002 194 −6.0 5.0 −0.5 38.8 0.853 3B OMP IPA 0.02 297 −9.3 1.7 −3.8 48.2 0.172 3B OMP IPA 0.002 185 −8.8 3.1 −2.8 41.0 0.346 3B Gad2 PEA 0.02 106 −12.2 1.1 −5.5 34.6 0.101 3B Gad2 PEA 0.002 84 −10.8 5.7 −2.5 38.2 0.542 3B Gad2 IPA 0.02 119 −8.5 7.1 −0.7 43.2 0.862 3B Gad2 IPA 0.002 62 −13.0 7.4 −2.8 40.1 0.582 3B DAT PEA 0.02 216 −6.0 5.3 −0.3 42.3 0.905 3B DAT PEA 0.002 118 −7.7 7.9 0.1 42.7 0.983 3B DAT IPA 0.02 184 −10.0 1.0 −4.5 37.9 0.111 3B DAT IPA 0.002 123 −13.1 0.8 −6.2 38.9 0.080 3B Pcdh21 PEA 0.02 199 −7.2 3.1 −2.1 36.8 0.428 3B Pcdh21 PEA 0.002 130 −12.7 2.4 −5.1 43.6 0.182 3B Pcdh21 IPA 0.02 241 −4.1 6.9 1.4 43.1 0.610 3B Pcdh21 IPA 0.002 176 −10.8 1.9 −4.4 42.4 0.166 4B OMP PEA 0.02 279 −2.1 15.8 6.9 76.1 0.133 4B OMP PEA 0.002 194 −3.5 18.8 7.6 78.7 0.178 4B OMP IPA 0.02 297 −2.6 11.9 4.7 63.6 0.207 4B OMP IPA 0.002 185 −3.7 14.5 5.4 62.7 0.245 4B Gad2 PEA 0.02 106 −36.1 3.1 −16.5 101.7 0.098 4B Gad2 PEA 0.002 84 −26.5 24.0 −1.2 116.2 0.922 4B Gad2 IPA 0.02 119 −28.6 7.7 −10.4 99.9 0.257 4B Gad2 IPA 0.002 62 −16.9 30.0 6.6 92.3 0.577 4B DAT PEA 0.02 216 −17.6 9.5 −4.1 100.7 0.555 4B DAT PEA 0.002 118 −19.5 22.3 1.4 114.5 0.893 4B DAT IPA 0.02 184 −27.3 3.0 −12.2 103.5 0.116 4B DAT IPA 0.002 123 −23.2 12.2 −5.5 100.5 0.541 4B Pcdh21 PEA 0.02 199 −4.3 24.7 10.2 103.7 0.165 4B Pcdh21 PEA 0.002 130 −12.9 17.9 2.5 88.7 0.745 4B Pcdh21 IPA 0.02 241 −10.0 20.2 5.1 119.0 0.504 4B Pcdh21 IPA 0.002 176 −5.8 20.8 7.5 89.2 0.267 5B OMP PEA 0.02 279 −5.1 12.3 3.6 74.2 0.419 5B OMP PEA 0.002 194 −8.6 15.2 3.3 83.9 0.583 5B OMP IPA 0.02 297 −4.2 14.2 5.0 80.8 0.289 5B OMP IPA 0.002 185 −8.3 16.4 4.1 85.3 0.517 5B Gad2 PEA 0.02 106 −12.9 17.6 2.4 79.2 0.757 5B Gad2 PEA 0.002 84 −15.5 18.3 1.4 77.8 0.867 5B Gad2 IPA 0.02 119 −6.6 22.9 8.2 81.4 0.276 5B Gad2 IPA 0.002 62 −15.5 18.3 1.4 66.6 0.869 5B DAT PEA 0.02 216 −8.6 21.2 6.3 111.0 0.407 5B DAT PEA 0.002 118 −21.7 15.9 −2.9 103.1 0.757 5B DAT IPA 0.02 184 −16.4 12.5 −2.0 99.4 0.789 5B DAT IPA 0.002 123 −27.8 6.3 −10.7 95.5 0.215 5B Pcdh21 PEA 0.02 199 −12.5 19.8 3.6 115.7 0.659 5B Pcdh21 PEA 0.002 130 −6.9 19.7 6.4 76.6 0.345 5B Pcdh21 IPA 0.02 241 −16.5 5.7 −5.4 88.2 0.337 5B Pcdh21 IPA 0.002 176 −3.7 21.4 8.8 82.9 0.166 6B OMP PEA 0.02 279 0.998 1.025 1.012 0.115 0.385 6B OMP PEA 0.002 194 0.994 1.034 1.014 0.142 0.864 6B OMP IPA 0.02 297 0.987 1.016 1.002 0.127 0.274 6B OMP IPA 0.002 185 0.985 1.023 1.004 0.133 0.708 6B Gad2 PEA 0.02 106 0.985 1.044 1.014 0.173 0.629 6B Gad2 PEA 0.002 84 0.982 1.047 1.014 0.166 0.531 6B Gad2 IPA 0.02 119 0.998 1.056 1.027 0.181 0.077 6B Gad2 IPA 0.002 62 0.994 1.077 1.035 0.189 0.479 6B DAT PEA 0.02 216 0.991 1.028 1.009 0.150 0.646 6B DAT PEA 0.002 118 0.983 1.039 1.011 0.167 0.472 6B DAT IPA 0.02 184 0.985 1.024 1.005 0.143 0.624 6B DAT IPA 0.002 123 0.979 1.033 1.006 0.166 0.680 6B Pcdh21 PEA 0.02 199 0.996 1.041 1.019 0.178 0.648 6B Pcdh21 PEA 0.002 130 0.963 1.020 0.992 0.180 0.216 6B Pcdh21 IPA 0.02 241 0.997 1.037 1.017 0.170 0.652 6B Pcdh21 IPA 0.002 176 1.003 1.050 1.026 0.172 0.159 7C Pre OMP PEA 0.02 279 0.984 1.011 0.998 0.111 0.703 7C Pre OMP PEA 0.002 194 0.980 1.016 0.998 0.129 0.834 7C Pre OMP IPA 0.02 297 0.997 1.026 1.011 0.128 0.135 7C Pre OMP IPA 0.002 185 0.992 1.030 1.011 0.122 0.253 7C Pre Gad2 PEA 0.02 106 1.300 1.407 1.354 0.289 4.011 E–24 7C Pre Gad2 PEA 0.002 84 1.270 1.389 1.330 0.282 1.935 E–18 7C Pre Gad2 IPA 0.02 119 1.296 1.407 1.351 0.322 4.059 E–24 7C Pre Gad2 IPA 0.002 62 1.304 1.449 1.376 0.315 4.941 E–16 7C Pre DAT PEA 0.02 216 1.258 1.323 1.291 0.245 3.173 E–43 7C Pre DAT PEA 0.002 118 1.220 1.304 1.262 0.230 8.205 E–23 7C Pre DAT IPA 0.02 184 1.222 1.287 1.255 0.226 1.910 E–35 7C Pre DAT IPA 0.002 123 1.201 1.302 1.251 0.276 6.661 E–17 7C Pre Pcdh21 PEA 0.02 199 1.264 1.351 1.308 0.339 1.512 E–32 7C Pre Pcdh21 PEA 0.002 130 1.247 1.347 1.297 0.300 1.512 E–22 7C Pre Pcdh21 IPA 0.02 241 1.292 1.357 1.325 0.273 6.790 E–55 7C Pre Pcdh21 IPA 0.002 176 1.281 1.355 1.318 0.258 1.024 E–39 7C Odor OMP PEA 0.02 279 0.990 1.021 1.005 0.131 0.492 7C Odor OMP PEA 0.002 194 0.997 1.032 1.015 0.128 0.104 7C Odor OMP IPA 0.02 297 0.988 1.019 1.004 0.135 0.635 7C Odor OMP IPA 0.002 185 0.984 1.021 1.002 0.120 0.812 7C Odor Gad2 PEA 0.02 106 1.475 1.630 1.552 0.415 1.008 E–26 7C Odor Gad2 PEA 0.002 84 1.506 1.693 1.600 0.448 1.009 E–21 7C Odor Gad2 IPA 0.02 119 1.508 1.670 1.589 0.469 7.588 E–29 7C Odor Gad2 IPA 0.002 62 1.491 1.652 1.571 0.350 9.631 E–23 7C Odor DAT PEA 0.02 216 1.405 1.495 1.450 0.338 7.929 E–50 7C Odor DAT PEA 0.002 118 1.424 1.564 1.494 0.382 1.060 E–26 7C Odor DAT IPA 0.02 184 1.413 1.522 1.467 0.382 2.147 E–39 7C Odor DAT IPA 0.002 123 1.407 1.555 1.481 0.400 3.559 E–24 7C Odor Pcdh21 PEA 0.02 199 1.416 1.507 1.462 0.351 5.482 E–53 7C Odor Pcdh21 PEA 0.002 130 1.460 1.595 1.528 0.405 6.719 E–32 7C Odor Pcdh21 IPA 0.02 241 1.435 1.529 1.482 0.397 2.219 E–56 7C Odor Pcdh21 IPA 0.002 176 1.464 1.587 1.526 0.427 1.386 E–39 7D OMP PEA 0.02 558 1.347 1.421 1.384 0.454 3.053 E–05 7D OMP PEA 0.002 388 1.413 1.496 1.454 0.394 0.017 7D OMP IPA 0.02 594 1.371 1.434 1.403 0.388 9.829 E–14 7D OMP IPA 0.002 370 1.275 1.353 1.314 0.359 3.323 E–06 7D Gad2 PEA 0.02 212 1.395 1.484 1.440 0.340 1.917 E–04 7D Gad2 PEA 0.002 168 1.402 1.514 1.458 0.384 1.931 E–04 7D Gad2 IPA 0.02 238 1.404 1.496 1.450 0.379 2.091 E–04 7D Gad2 IPA 0.002 124 1.415 1.523 1.469 0.335 1.401 E–03 7D DAT PEA 0.02 432 1.338 1.393 1.365 0.292 8.842 E–06 7D DAT PEA 0.002 236 1.327 1.403 1.365 0.295 4.632 E–05 7D DAT IPA 0.02 368 1.319 1.379 1.349 0.294 8.927 E–05 7D DAT IPA 0.002 246 1.307 1.387 1.347 0.309 2.816 E–04 7D Pcdh21 PEA 0.02 398 1.348 1.409 1.378 0.339 8.010 E–03 7D Pcdh21 PEA 0.002 260 1.360 1.441 1.400 0.344 5.501 E–03 7D Pcdh21 IPA 0.02 482 1.363 1.416 1.390 0.314 3.800 E–03 7D Pcdh21 IPA 0.002 352 1.369 1.433 1.401 0.316 8.931 E–04
Extended Data Figure 7-1
Corrections for white-noise background in the power spectra. Representative power spectra of calcium signals from GCaMP-expressing OMP-Cre (A1), Gad2-Cre (A2), DAT-Cre (A3), and Pcdh21-Cre (A4) mice are presented. In each subpanel, the left and right columns represent the power spectra before and after correction for the white-noise background, respectively. The spectra in the upper and lower rows in the sub-panel are those from the prestimulation and the odor stimulation periods, respectively. The original plots in each mice (left columns in subpanels) are the same as those shown in Figure 7B. The background baseline was defined for each power spectrum as a mean value between 6 and 9 Hz. Then, these background values were subtracted from the original spectra. Download Figure 7-1, EPS file.
In this issue
