Article Figures & Data
Figures
- Cover Figure
Region-specific adenylyl cyclase 1 (AC1) loss of function differentially affects both patterning and sensorimotor behaviors in mice. AC1 is expressed at all levels of the somatosensory pathway and plays a major role in refinement and patterning of topographic sensory maps. Cortex-specific AC1 loss of function (CxAC1KO mice) does not affect barrel patterning and activation of specific barrels corresponding to stimulated whiskers and does not impair sensorimotor behaviors. While global (AC1KO) and thalamus-specific (ThAC1KO) AC1 loss of function leads to absence of barrel patterns, selective whisker stimulation activates topographically aligned cortical loci. Despite functional topography of the whisker-barrel cortex, sensorimotor and social behaviors are impaired, indicating the importance of patterning of topographical sensory maps in the neocortex.
- Figure 1
Barrel patterns and whisker-related activation in the barrel cortex. A, B, VGLUT-2 (green, for TCAs) and Neun (blue, for neurons) double immunostaining reveals that both the TCA terminal segregation and cellular organization into barrels appear normal in CxAC1KO cortex (A). In the ThAC1KO cortex, cellular patterning is absent (note the rather uniform distribution of the Neun-labeled blue cells) and there is some patterning of the TCA terminals (green; B). C, D, VGLUT-2 (blue) and c-fos (pink) immunolabeling in the barrel cortex of a B6 (control, C) and AC1KO (D) mice following row C whisker activation. Note that in the AC1KO cortex, a topographically appropriate zone is activated, although there is no VGLUT-2-related patterning. E, F, VGLUT-2 (blue) and c-fos (pink) immunolabeling in the barrel cortex of exemplary CxAC1KO and ThAC1KO mice following row C whisker activation. Note that the activity pattern in the CxAC1KO cortex is similar to B6 control, while that of the ThAC1KO cortex is similar to the AC1KO cortex. Scale bar, 200 μm.
- Figure 2
Sensorimotor behavior comparisons. A, B, Gap crossing and edge approach tests. WT mice crossed longer gap distances than WC or AC1KO mice, F(2,23) = 8.276, p = 0.0022. ThAC1KO mice crossed shorter gap distances than control AC1flox/− (AC1ff) mice, F(2,21) = 12.70, p = 0.0002, while CxAC1KO mice performed similar to controls. In the edge approach test, WC and AC1KO mice were significantly impaired compared with WT controls, F(2,23) = 14.071, p = 0.0001. Likewise, ThAC1KO mice reached significantly shorter distances compared with CxAC1KO or AC1ff controls, F(2,21) = 10.971, p = 0.0005. C, D, Swimming and paw sensation tests. In the swimming test, AC1KO and WC mice showed a similar level of struggling, F(2,23) = 0.190, p = 0.8283, but significantly shorter periods of floating than WT controls, F(2,23) = 6.572, p = 0.0055. Cx, ThAC1KO, and AC1ff controls all showed a similar level of struggling, F(2,21) = 2.452, p = 0.1104, while ThAC1KO mice floated significantly shorter time than either CxAC1KO or AC1ff controls, F(2,21) = 11.754, p = 0.0004. In the sticky paper test, AC1KO mice displayed higher latencies for licking off the paper stuck on their hind paw, F(2,23) = 8.175, p = 0.0021. ThAC1KO mice also showed higher latencies licking the paper compared with either CxAC1KO or AC1ff mice, F(2,21) = 4.636, p = 0.0215. All data are expressed as mean ± SEM * indicates significant differences between strains.
- Figure 3
Whisking and object recognition tests. A, Whisking behavior comparisons between AC1KO, WT, and WC mice. AC1KO mice showed a lower ratio of active whisking whisker contact with an object compared with WT mice; t(16) = 3.4495, p = 0.0063. WT mice displayed a clear difference in vibration frequencies between symmetry (nontouch) and active (touch) whisking—the frequencies of active whisking were higher than symmetry whisking—while AC1KO mice showed similar frequencies between symmetry and active whisking. This was supported by a two-way ANOVA; strain: F(1,14) = 7.007, p = 0.0191, whisking type: F(1,14) = 25.373, p = 0.0002, and the interaction between strain and whisking type: F(1,14) = 16.174, p = 0.0013. B, Whisking comparisons between CxAC1KO, ThAC1KO, and control AC1flox/− (AC1ff) mice. ThAC1KO mice showed a lower ratio of active whisking during object contact with whiskers than CxAC1KO or their controls (AC1ff); F(2,21) = 9.223, p = 0.0013. CxAC1KO and AC1ff mice displayed a clear difference in vibration frequencies between symmetry (nontouch) and active (touch) whisking—the frequencies of active whisking were higher than symmetry whisking—while ThAC1KO mice show similar frequencies between symmetry and active whisking. This was supported by a two-way ANOVA; strain: F(2,21) = 18.42, p = 0.0035, whisking type: F(1,21) = 72.46, p = 0.002, and the interaction between strain and whisking type: F(2,21) = 17.273, p = 0.0054. C, Texture and object discrimination. In the object discrimination test, there were no significant strain differences throughout trials, while the preference ratio on trials 2 and 3 were higher than trial 1; a two-way ANOVA: strain: F(2,23) = 1.416, p = 0.263; trial: F(2,46) = 67.984, p = 0.0001; and strain × trial: F(2,46) = 1.668, p = 0.1738. However, in the texture discrimination test, WT mice clearly showed a higher preference toward an unfamiliar textured object than familiar one on trials 2 and 3, while AC1KO and WC mice did not show significantly lower preference than WT control on trial 1, but did on trials 2 and 3; a two-way ANOVA: strain: F(2,23) = 26.438, p < 0.0001; trial: F(2,46) = 0.3997, p = 0.634; and strain × trial: F(2,46) = 13.910, p < 0.0001. In the object-discrimination test, there were no significant strain differences between CxAC1KO, AC1ff, and ThAC1KO throughout trials, while the preference ratio on trials 2 and 3 were higher than on trial 1; a two-way ANOVA: strain: F(2,21) = 0.24, n.s.; trial: F(2,42) = 49.57, p = 0.0003; and strain × trial: F(2,42) = 0.38, n.s. In the texture-discrimination test, CxAC1KO and AC1ff mice displayed a higher preference toward an unfamiliar textured object than familiar one on trials 2 and 3, while ThAC1KO mice showed similar preference throughout all three trials; a two-way ANOVA: strain: F(2,21) = 12.45, p = 0.0087; trial: F(2,42) = 13.96, p = 0.0075; and strain × trial: F(2,42) = 9.59, p = 0.0082. All data are expressed as mean ± SEM. * indicates significant differences between strains, # indicates significant differences between trials.
- Figure 4
General motor behavior. A, B, In the wire-hanging test, there were no significant differences between any of the strains. C, D, In open-field exploration tests, there was no significant difference in locomotion, F(2,23) = 0.317, p = 0.7316 and F(2,21) = 1.821, p = 0.1865.
- Figure 5
Social behavior. A, B, In the social behavior test, there was no difference in the duration of social contacts, F(2,23) = 0.084, p = 0.9201. However, AC1KO mice displayed a notable increment of flight response to the intruder’s contacts from the front, F(2,23) = 7.662, p = 0.0028, and back sides, F(2,23) = 48.434, p = 0.0001, compared with WT or WC mice. C, D, In comparisons of the second group of mice, there was no difference in the duration of social contacts, F(2,21) = 5.896, p = 0.093. However, ThAC1KO mice displayed a greater flight response to the intruder’s contacts from the front, F(2,21) = 25.792, p = 0.0001, and back sides, F(2,21) = 46.706, p < 0.0001, compared with either CxAC1KO or AC1ff mice. All data are expressed as mean ± SEM * indicates significant differences between strains, # indicates significant differences between trials.
Tables
Data structure Type of test Power Fig. 2. gap distances, AC1KO, WC, and WT Normal distribution One–way ANOVA 1.0000 Fig. 2. gap distances, ThAC1KO, CxAC1KO, and control Normal distribution One–way ANOVA 1.0000 Fig. 2. edge approach, AC1KO, WC, and WT Normal distribution One–way ANOVA 1.0000 Fig. 2. edge approach, ThAC1KO, CxAC1KO, and control Normal distribution One–way ANOVA 1.0000 Fig. 2. swimming, struggling AC1KO, WC, and WT Normal distribution One–way ANOVA 0.8000 Fig. 2. swimming, floating AC1KO, WC, and WT Normal distribution One–way ANOVA 0.9999 Fig. 2. swimming, struggling ThAC1KO, CxAC1KO, and control Normal distribution One–way ANOVA 0.8507 Fig. 2. swimming, floating ThAC1KO, CxAC1KO, and control Normal distribution One–way ANOVA 1.0000 Fig. 2. sticky paper, AC1KO, WC, and WT Normal distribution One–way ANOVA 1.0000 Fig. 2. sticky paper, ThAC1KO, CxAC1KO, and control Normal distribution One–way ANOVA 0.9976 Fig. 3. active whisking frequency, AC1KO, WC, and WT Normal distribution Two–way ANOVA, strain 0.8831 Normal distribution Two–way ANOVA, type 1.0000 Normal distribution Two–way ANOVA, interaction 1.0000 Fig. 3. active whisking frequency, ThAC1KO, CxAC1KO, and control Normal distribution Two–way ANOVA, strain 1.0000 Normal distribution Two–way ANOVA, type 1.0000 Normal distribution Two–way ANOVA, interaction 1.0000 Fig. 3. object discrimination, AC1KO, WC, and WT Normal distribution Two–way ANOVA, strain 1.0000 Normal distribution Two–way ANOVA, trial 0.8100 Normal distribution Two–way ANOVA, interaction 1.0000 Fig. 3. object discrimination, ThAC1KO, CxAC1KO, and control Normal distribution Two–way ANOVA, strain 0.9485 Normal distribution Two–way ANOVA, trial 1.0000 Normal distribution Two–way ANOVA, interaction 0.9999 Fig. 3. texture discrimination, AC1KO, WC, and WT Normal distribution Two–way ANOVA, strain 1.0000 Normal distribution Two–way ANOVA, trial 1.0000 Normal distribution Two–way ANOVA, interaction 1.0000 Fig. 3. texture discrimination, ThAC1KO, CxAC1KO, and control Normal distribution Two–way ANOVA, strain 0.8090 Normal distribution Two–way ANOVA, trial 1.0000 Normal distribution Two–way ANOVA, interaction 0.8470 Fig. 4. open field locomotion, AC1KO, WC, and WT Normal distribution One–way ANOVA 0.8003 Fig. 4. open field locomotion, ThAC1KO, CxAC1KO, and control Normal distribution One–way ANOVA 0.9649 Fig. 5. social contacts AC1KO, WC, and WT Normal distribution One–way ANOVA 0.8008 Fig. 5. flight to front approach, AC1KO, WC, and WT Normal distribution One–way ANOVA 1.0000 Fig. 5. flight to back approach, ThAC1KO, CxAC1KO, and control Normal distribution One–way ANOVA 1.0000 Fig. 5. social contacts, ThAC1KO, CxAC1KO, and control Normal distribution One–way ANOVA 0.9999 Fig. 5. flight to front approach, ThAC1KO, CxAC1KO, and control Normal distribution One–way ANOVA 1.0000 Fig. 5. flight to back approach, ThAC1KO, CxAC1KO, and control Normal distribution One–way ANOVA 1.0000
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