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<t>DREADD</t> expression in noradrenergic <t>neurons.</t> <t>DREADD/mCherry-inducing</t> viral vectors were administered to the NTS and LC of DBH-CRE mice. (A) mCherry-expressing neurons (red) co-labeled with TH (green) in the NTS. Few off-target mCherry-positive neurons were observed in the area postrema (AP). (B) Over 90% of the mCherry-expressing neurons in the NTS were immunopositive for TH. ( C ) mCherry-expressing neurons co-labeled with TH in the LC. ( D ) Over 90% of the mCherry-expressing neurons in the LC were immunopositive for TH. ( E ) cFos staining in the LC 90 min following DCZ injection IP in mice expressing the mCherry control virus (Control) or the Gq-DREADD. ( F ) cFOS was expressed in a significantly greater number of LC neurons following DCZ injection in the DREADD-expressing compared to the control virus-expressing mice. Scale bars = 100 μm.

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1) Product Images from "Norepinephrine neurons in the locus coeruleus and the nucleus of the solitary tract drive different stress-related behavioral outputs in mice"

Article Title: Norepinephrine neurons in the locus coeruleus and the nucleus of the solitary tract drive different stress-related behavioral outputs in mice

Journal: Neurobiology of Stress

doi: 10.1016/j.ynstr.2026.100802

DREADD expression in noradrenergic neurons. DREADD/mCherry-inducing viral vectors were administered to the NTS and LC of DBH-CRE mice. (A) mCherry-expressing neurons (red) co-labeled with TH (green) in the NTS. Few off-target mCherry-positive neurons were observed in the area postrema (AP). (B) Over 90% of the mCherry-expressing neurons in the NTS were immunopositive for TH. ( C ) mCherry-expressing neurons co-labeled with TH in the LC. ( D ) Over 90% of the mCherry-expressing neurons in the LC were immunopositive for TH. ( E ) cFos staining in the LC 90 min following DCZ injection IP in mice expressing the mCherry control virus (Control) or the Gq-DREADD. ( F ) cFOS was expressed in a significantly greater number of LC neurons following DCZ injection in the DREADD-expressing compared to the control virus-expressing mice. Scale bars = 100 μm.

Figure Legend Snippet: DREADD expression in noradrenergic neurons. DREADD/mCherry-inducing viral vectors were administered to the NTS and LC of DBH-CRE mice. (A) mCherry-expressing neurons (red) co-labeled with TH (green) in the NTS. Few off-target mCherry-positive neurons were observed in the area postrema (AP). (B) Over 90% of the mCherry-expressing neurons in the NTS were immunopositive for TH. ( C ) mCherry-expressing neurons co-labeled with TH in the LC. ( D ) Over 90% of the mCherry-expressing neurons in the LC were immunopositive for TH. ( E ) cFos staining in the LC 90 min following DCZ injection IP in mice expressing the mCherry control virus (Control) or the Gq-DREADD. ( F ) cFOS was expressed in a significantly greater number of LC neurons following DCZ injection in the DREADD-expressing compared to the control virus-expressing mice. Scale bars = 100 μm.

Techniques Used: Expressing, Labeling, Staining, Injection, Control, Virus

Chemogenetic activation of PVN-projecting NE neurons induced multiple stress behaviors in male and female mice. (A) A Cre-dependent, Gq-DREADD-expressing retrograde AAV was injected into the PVN of DBH-Cre mice. (B) Noradrenergic neurons in the NTS and the LC were labeled with tyrosine hydroxylase immunofluorescence (TH-ir); sparse expression of mCherry immunofluorescence (mCherry-ir) was observed in the LC and NTS. (C) mCherry-immunopositive axons were visible in the PVN. (D) No mCherry immunofluorescence was detected in the rostral ventro-lateral medulla (RVLM). (E, F) DCZ activation of NE neurons retrogradely labeled from the PVN decreased the exploratory behaviors of walking (E) and rearing (F). (G, H) DCZ caused a trend toward an increase in the time spent grooming (G) and caused a significant increase in immobility (H). (I) Regression analysis revealed a strong correlation between time spent in grooming and immobility behaviors in these mice. (J) Food intake was not significantly affected by Gq-DREADD activation of PVN-projecting NE neurons. White arrows in panel B indicate co-localization of mCherry and TH immunofluorescence.

Figure Legend Snippet: Chemogenetic activation of PVN-projecting NE neurons induced multiple stress behaviors in male and female mice. (A) A Cre-dependent, Gq-DREADD-expressing retrograde AAV was injected into the PVN of DBH-Cre mice. (B) Noradrenergic neurons in the NTS and the LC were labeled with tyrosine hydroxylase immunofluorescence (TH-ir); sparse expression of mCherry immunofluorescence (mCherry-ir) was observed in the LC and NTS. (C) mCherry-immunopositive axons were visible in the PVN. (D) No mCherry immunofluorescence was detected in the rostral ventro-lateral medulla (RVLM). (E, F) DCZ activation of NE neurons retrogradely labeled from the PVN decreased the exploratory behaviors of walking (E) and rearing (F). (G, H) DCZ caused a trend toward an increase in the time spent grooming (G) and caused a significant increase in immobility (H). (I) Regression analysis revealed a strong correlation between time spent in grooming and immobility behaviors in these mice. (J) Food intake was not significantly affected by Gq-DREADD activation of PVN-projecting NE neurons. White arrows in panel B indicate co-localization of mCherry and TH immunofluorescence.

Techniques Used: Activation Assay, Expressing, Injection, Labeling, Immunofluorescence

Chemogenetic activation of NE neurons in male and female mice. ( A ) Experimental design: A Gq-DREADD-expressing AAV was injected bilaterally into the NTS of DBH-Cre mice and the mice were subjected subsequently to I.P. injections of CNO and vehicle at a two-week interval, in a cross-over design. Inset: Viral expression of the DREADD was confirmed by confocal imaging of mCherry fluorescence in the NTS. ( B ) Chemogenetic stimulation of NE neurons in the NTS activated the HPA axis, as indicated by an increase in serum corticosterone in both males and females. ( C, D ) Vehicle-injected control males spent more time walking (C) and rearing (D) than control females; chemogenetic activation of NTS NE neurons significantly decreased walking and rearing in both males and females and abolished the sex difference in the response. ( E ) Grooming behavior after vehicle injection did not differ between males and females; CNO caused a decrease in the time spent grooming in both sexes. ( F ) CNO caused a robust increase in the time both males and females spent in immobility, with no differences between sexes.

Figure Legend Snippet: Chemogenetic activation of NE neurons in male and female mice. ( A ) Experimental design: A Gq-DREADD-expressing AAV was injected bilaterally into the NTS of DBH-Cre mice and the mice were subjected subsequently to I.P. injections of CNO and vehicle at a two-week interval, in a cross-over design. Inset: Viral expression of the DREADD was confirmed by confocal imaging of mCherry fluorescence in the NTS. ( B ) Chemogenetic stimulation of NE neurons in the NTS activated the HPA axis, as indicated by an increase in serum corticosterone in both males and females. ( C, D ) Vehicle-injected control males spent more time walking (C) and rearing (D) than control females; chemogenetic activation of NTS NE neurons significantly decreased walking and rearing in both males and females and abolished the sex difference in the response. ( E ) Grooming behavior after vehicle injection did not differ between males and females; CNO caused a decrease in the time spent grooming in both sexes. ( F ) CNO caused a robust increase in the time both males and females spent in immobility, with no differences between sexes.

Techniques Used: Activation Assay, Expressing, Injection, Imaging, Fluorescence, Control

Chemogenetic activation of NE neurons in the LC in male and female mice. (A) Experimental design: A Gq-DREADD-expressing AAV was injected bilaterally into the LC of DBH-Cre mice, then the mice were injected IP with DCZ and vehicle at a two-week interval, in a cross-over design. Inset: Viral expression of the DREADD was confirmed by confocal imaging of mCherry fluorescence in the LC. (B ) Chemogenetic stimulation of NE neurons in the LC activated the HPA axis, as indicated by an increase in serum corticosterone in both males and females. ( C-F) DCZ injections IP caused a decrease in the time spent walking (C) and rearing (D) and an increase in the time spent grooming (E), but had no effect on the average time spent in immobility (E). (F) There was a moderate but significant decrease in food intake in response to DCZ. There was no significant effect of sex on any of the behavioral measures.

Figure Legend Snippet: Chemogenetic activation of NE neurons in the LC in male and female mice. (A) Experimental design: A Gq-DREADD-expressing AAV was injected bilaterally into the LC of DBH-Cre mice, then the mice were injected IP with DCZ and vehicle at a two-week interval, in a cross-over design. Inset: Viral expression of the DREADD was confirmed by confocal imaging of mCherry fluorescence in the LC. (B ) Chemogenetic stimulation of NE neurons in the LC activated the HPA axis, as indicated by an increase in serum corticosterone in both males and females. ( C-F) DCZ injections IP caused a decrease in the time spent walking (C) and rearing (D) and an increase in the time spent grooming (E), but had no effect on the average time spent in immobility (E). (F) There was a moderate but significant decrease in food intake in response to DCZ. There was no significant effect of sex on any of the behavioral measures.

Techniques Used: Activation Assay, Expressing, Injection, Imaging, Fluorescence

Effects of manipulation of NTS-NE neurons on sickness-like behaviors. (A) Excitatory Gq-DREADD-expressing, inhibitory Gi-DREADD-expressing, or mCherry control (CTRL) viral vectors were injected bilaterally into the NTS of DBH-Cre mice, after which they were subjected to a battery of sickness-related behavioral tests. ( B ) mCherry expression in the NTS following control (CTRL), Gq-DREADD- (Gq), and Gi-DREADD (Gi)-expressing virus injection. ( C-E ) In the open field test, DCZ injection in Gq-DREADD-expressing mice caused a significant decrease in the total distance travelled (C) and the time spent in movement (D), and an increase in the time spent in immobility (E) compared to both Gi-DREADD-expressing and CTRL mice. ( F ) Time spent in the center of the open field did not differ among groups. ( G ) Food intake was significantly decreased by DCZ in the Gq-DREADD-injected mice compared to the Gi-DREADD-injected and CTRL mice. ( H ) Using the von Frey test, half the Gq-DREADD-injected mice were not responsive to mechanical stimuli, which was not observed in the Gi-DREADD-injected or CTRL mice. ( I ) Among the mechano-responsive mice, the von Frey threshold was significantly reduced in the Gq-DREADD-injected mice compared to the Gi-DREADD-injected and CTRL mice. ∗∗p < 0.01, ∗∗∗p < 0.001.

Figure Legend Snippet: Effects of manipulation of NTS-NE neurons on sickness-like behaviors. (A) Excitatory Gq-DREADD-expressing, inhibitory Gi-DREADD-expressing, or mCherry control (CTRL) viral vectors were injected bilaterally into the NTS of DBH-Cre mice, after which they were subjected to a battery of sickness-related behavioral tests. ( B ) mCherry expression in the NTS following control (CTRL), Gq-DREADD- (Gq), and Gi-DREADD (Gi)-expressing virus injection. ( C-E ) In the open field test, DCZ injection in Gq-DREADD-expressing mice caused a significant decrease in the total distance travelled (C) and the time spent in movement (D), and an increase in the time spent in immobility (E) compared to both Gi-DREADD-expressing and CTRL mice. ( F ) Time spent in the center of the open field did not differ among groups. ( G ) Food intake was significantly decreased by DCZ in the Gq-DREADD-injected mice compared to the Gi-DREADD-injected and CTRL mice. ( H ) Using the von Frey test, half the Gq-DREADD-injected mice were not responsive to mechanical stimuli, which was not observed in the Gi-DREADD-injected or CTRL mice. ( I ) Among the mechano-responsive mice, the von Frey threshold was significantly reduced in the Gq-DREADD-injected mice compared to the Gi-DREADD-injected and CTRL mice. ∗∗p < 0.01, ∗∗∗p < 0.001.

Techniques Used: Expressing, Control, Injection, Battery, Virus

Image Search Results

Journal: Neurobiology of Stress

Article Title: Norepinephrine neurons in the locus coeruleus and the nucleus of the solitary tract drive different stress-related behavioral outputs in mice

doi: 10.1016/j.ynstr.2026.100802

Figure Lengend Snippet: DREADD expression in noradrenergic neurons. DREADD/mCherry-inducing viral vectors were administered to the NTS and LC of DBH-CRE mice. (A) mCherry-expressing neurons (red) co-labeled with TH (green) in the NTS. Few off-target mCherry-positive neurons were observed in the area postrema (AP). (B) Over 90% of the mCherry-expressing neurons in the NTS were immunopositive for TH. ( C ) mCherry-expressing neurons co-labeled with TH in the LC. ( D ) Over 90% of the mCherry-expressing neurons in the LC were immunopositive for TH. ( E ) cFos staining in the LC 90 min following DCZ injection IP in mice expressing the mCherry control virus (Control) or the Gq-DREADD. ( F ) cFOS was expressed in a significantly greater number of LC neurons following DCZ injection in the DREADD-expressing compared to the control virus-expressing mice. Scale bars = 100 μm.

Article Snippet: In addition, a virus that induces the expression of mCherry without a DREADD (AAV9-hSyn-DIO-mCherry, 0.66 ∗ 3 13 GC/mL, Addgene 50459-AAV9) was injected bilaterally into the NTS as a control (CTRL).

Techniques: Expressing, Labeling, Staining, Injection, Control, Virus

Journal: Neurobiology of Stress

Article Title: Norepinephrine neurons in the locus coeruleus and the nucleus of the solitary tract drive different stress-related behavioral outputs in mice

doi: 10.1016/j.ynstr.2026.100802

Figure Lengend Snippet: Chemogenetic activation of PVN-projecting NE neurons induced multiple stress behaviors in male and female mice. (A) A Cre-dependent, Gq-DREADD-expressing retrograde AAV was injected into the PVN of DBH-Cre mice. (B) Noradrenergic neurons in the NTS and the LC were labeled with tyrosine hydroxylase immunofluorescence (TH-ir); sparse expression of mCherry immunofluorescence (mCherry-ir) was observed in the LC and NTS. (C) mCherry-immunopositive axons were visible in the PVN. (D) No mCherry immunofluorescence was detected in the rostral ventro-lateral medulla (RVLM). (E, F) DCZ activation of NE neurons retrogradely labeled from the PVN decreased the exploratory behaviors of walking (E) and rearing (F). (G, H) DCZ caused a trend toward an increase in the time spent grooming (G) and caused a significant increase in immobility (H). (I) Regression analysis revealed a strong correlation between time spent in grooming and immobility behaviors in these mice. (J) Food intake was not significantly affected by Gq-DREADD activation of PVN-projecting NE neurons. White arrows in panel B indicate co-localization of mCherry and TH immunofluorescence.

Techniques: Activation Assay, Expressing, Injection, Labeling, Immunofluorescence

Journal: Neurobiology of Stress

Article Title: Norepinephrine neurons in the locus coeruleus and the nucleus of the solitary tract drive different stress-related behavioral outputs in mice

doi: 10.1016/j.ynstr.2026.100802

Figure Lengend Snippet: Chemogenetic activation of NE neurons in male and female mice. ( A ) Experimental design: A Gq-DREADD-expressing AAV was injected bilaterally into the NTS of DBH-Cre mice and the mice were subjected subsequently to I.P. injections of CNO and vehicle at a two-week interval, in a cross-over design. Inset: Viral expression of the DREADD was confirmed by confocal imaging of mCherry fluorescence in the NTS. ( B ) Chemogenetic stimulation of NE neurons in the NTS activated the HPA axis, as indicated by an increase in serum corticosterone in both males and females. ( C, D ) Vehicle-injected control males spent more time walking (C) and rearing (D) than control females; chemogenetic activation of NTS NE neurons significantly decreased walking and rearing in both males and females and abolished the sex difference in the response. ( E ) Grooming behavior after vehicle injection did not differ between males and females; CNO caused a decrease in the time spent grooming in both sexes. ( F ) CNO caused a robust increase in the time both males and females spent in immobility, with no differences between sexes.

Techniques: Activation Assay, Expressing, Injection, Imaging, Fluorescence, Control

Journal: Neurobiology of Stress

Article Title: Norepinephrine neurons in the locus coeruleus and the nucleus of the solitary tract drive different stress-related behavioral outputs in mice

doi: 10.1016/j.ynstr.2026.100802

Figure Lengend Snippet: Chemogenetic activation of NE neurons in the LC in male and female mice. (A) Experimental design: A Gq-DREADD-expressing AAV was injected bilaterally into the LC of DBH-Cre mice, then the mice were injected IP with DCZ and vehicle at a two-week interval, in a cross-over design. Inset: Viral expression of the DREADD was confirmed by confocal imaging of mCherry fluorescence in the LC. (B ) Chemogenetic stimulation of NE neurons in the LC activated the HPA axis, as indicated by an increase in serum corticosterone in both males and females. ( C-F) DCZ injections IP caused a decrease in the time spent walking (C) and rearing (D) and an increase in the time spent grooming (E), but had no effect on the average time spent in immobility (E). (F) There was a moderate but significant decrease in food intake in response to DCZ. There was no significant effect of sex on any of the behavioral measures.

Techniques: Activation Assay, Expressing, Injection, Imaging, Fluorescence

Journal: Neurobiology of Stress

Article Title: Norepinephrine neurons in the locus coeruleus and the nucleus of the solitary tract drive different stress-related behavioral outputs in mice

doi: 10.1016/j.ynstr.2026.100802

Figure Lengend Snippet: Effects of manipulation of NTS-NE neurons on sickness-like behaviors. (A) Excitatory Gq-DREADD-expressing, inhibitory Gi-DREADD-expressing, or mCherry control (CTRL) viral vectors were injected bilaterally into the NTS of DBH-Cre mice, after which they were subjected to a battery of sickness-related behavioral tests. ( B ) mCherry expression in the NTS following control (CTRL), Gq-DREADD- (Gq), and Gi-DREADD (Gi)-expressing virus injection. ( C-E ) In the open field test, DCZ injection in Gq-DREADD-expressing mice caused a significant decrease in the total distance travelled (C) and the time spent in movement (D), and an increase in the time spent in immobility (E) compared to both Gi-DREADD-expressing and CTRL mice. ( F ) Time spent in the center of the open field did not differ among groups. ( G ) Food intake was significantly decreased by DCZ in the Gq-DREADD-injected mice compared to the Gi-DREADD-injected and CTRL mice. ( H ) Using the von Frey test, half the Gq-DREADD-injected mice were not responsive to mechanical stimuli, which was not observed in the Gi-DREADD-injected or CTRL mice. ( I ) Among the mechano-responsive mice, the von Frey threshold was significantly reduced in the Gq-DREADD-injected mice compared to the Gi-DREADD-injected and CTRL mice. ∗∗p < 0.01, ∗∗∗p < 0.001.

Techniques: Expressing, Control, Injection, Battery, Virus

( A ) Percentage of BrdU-labeled cells that are also Dock10+ at different time points following BrdU injections (in days). ( B ) Representative section showing BrdU+ labeling (green) and Dock10+ labeling (red) in Dock10 cre;Ai9 mice. Calibration bars: 10 μm. ( C ) Schematic representation of the pattern separation paradigm. All mice (males n = 17, females n = 27) received DCZ i.p. injections 30 min before the beginning of their LS trials. ( D ) Percentage of mice reaching criterion in (S) and percentage that were not successful (NS) after 14 days of training in S configuration of LS. A majority of mice never succeeded in reaching the criterion in the Cre+ DCZ group ( z = 2.00, p = 0.0453, z -test—Group NS). ( E ) Percentage of mice reaching the criterion in S trials across days represented as the cumulative probability including all successful and not successful mice in the group. ( F ) Density of c-Fos+ GCs in the dorsal DG across the two groups. A significantly reduced density was observed in the Cre+ DCZ group compared to controls (unpaired t -test, t (9) = 2.30, p = 0.047). ( G ) Density of c-Fos+ GCs in the dorsal CA3 across the two groups. A significant reduction was observed in the Cre+ DCZ group compared to the control groups (unpaired t -test, t (9) = 4.93, p = 0.0008). ( H ) Blade-specific distribution of c-Fos+ GCs in the IB and SB of the dorsal DG in mice performing LS separation. Two-way ANOVA (unbalanced, Type II) revealed a significant main effect of blade ( F (1,16) = 315.83, p = 5.87 × 10 − ¹²), but no significant main effect of group ( F (1,16) = 0, p = 1) and no significant group × blade interaction ( F (1,16) = 0.040, p = 0.845). ( I ) Example of dorsal DG section from Dock10-cre:hM4Di mice after LS training. c-Fos+ neurons are labeled in red. Calibration bars: 100 μm. ( J ) Spatial distribution of c-Fos+ along the hilar to outer radial granule cell layer (GCL) axis of the DG (0–120 μm), with 0 μm indicating location at the subgranular zone (SGZ). ( K ) Cartoon illustrating the distribution of activity labeled GCs in the SB and IB blades of the DG. GCs are depicted in gray, while red circles represent activated cells in the DG. The overall distribution of neurons in mice in which mGCs were inhibited (Cre+ DCZ) and which did not reach criterion was not different from controls even though the total density of activity labeled neurons was lower.

Journal: eLife

Article Title: Subregional activity in the dentate gyrus is amplified during elevated cognitive demands

doi: 10.7554/eLife.109611

Figure Lengend Snippet: ( A ) Percentage of BrdU-labeled cells that are also Dock10+ at different time points following BrdU injections (in days). ( B ) Representative section showing BrdU+ labeling (green) and Dock10+ labeling (red) in Dock10 cre;Ai9 mice. Calibration bars: 10 μm. ( C ) Schematic representation of the pattern separation paradigm. All mice (males n = 17, females n = 27) received DCZ i.p. injections 30 min before the beginning of their LS trials. ( D ) Percentage of mice reaching criterion in (S) and percentage that were not successful (NS) after 14 days of training in S configuration of LS. A majority of mice never succeeded in reaching the criterion in the Cre+ DCZ group ( z = 2.00, p = 0.0453, z -test—Group NS). ( E ) Percentage of mice reaching the criterion in S trials across days represented as the cumulative probability including all successful and not successful mice in the group. ( F ) Density of c-Fos+ GCs in the dorsal DG across the two groups. A significantly reduced density was observed in the Cre+ DCZ group compared to controls (unpaired t -test, t (9) = 2.30, p = 0.047). ( G ) Density of c-Fos+ GCs in the dorsal CA3 across the two groups. A significant reduction was observed in the Cre+ DCZ group compared to the control groups (unpaired t -test, t (9) = 4.93, p = 0.0008). ( H ) Blade-specific distribution of c-Fos+ GCs in the IB and SB of the dorsal DG in mice performing LS separation. Two-way ANOVA (unbalanced, Type II) revealed a significant main effect of blade ( F (1,16) = 315.83, p = 5.87 × 10 − ¹²), but no significant main effect of group ( F (1,16) = 0, p = 1) and no significant group × blade interaction ( F (1,16) = 0.040, p = 0.845). ( I ) Example of dorsal DG section from Dock10-cre:hM4Di mice after LS training. c-Fos+ neurons are labeled in red. Calibration bars: 100 μm. ( J ) Spatial distribution of c-Fos+ along the hilar to outer radial granule cell layer (GCL) axis of the DG (0–120 μm), with 0 μm indicating location at the subgranular zone (SGZ). ( K ) Cartoon illustrating the distribution of activity labeled GCs in the SB and IB blades of the DG. GCs are depicted in gray, while red circles represent activated cells in the DG. The overall distribution of neurons in mice in which mGCs were inhibited (Cre+ DCZ) and which did not reach criterion was not different from controls even though the total density of activity labeled neurons was lower.

Article Snippet: Strain, strain background ( Mus musculus ) , R26-LSL-Gi-DREADD (hM4Di) , Jackson Laboratory , JAX 026219 , Chemogenetic inhibition model.

Techniques: Labeling, Control, Activity Assay

( A ) Schematic representation of the pattern separation paradigm. Prior to the onset of the training, mice (males n = 17, females n = 16) were provided tamoxifen-containing food ad libitum in the home cage for 7 weeks to activate TAM inducible Cre recombinase (Ascl1-CreER T2 ) in abDGCs. All mice received DCZ (deschloroclozapine, 50 μg kg −1 ) or DMSO injections 30 min before the beginning of LS trials. ( B ) Example sections from Ascl1-CreER T2 ; hM4Di mice with immunohistochemical localization of the HA tagged DREADD (top) and after LS training with c-Fos+ neurons labeled in red. Calibration bars: 140 μm. ( C ) Percentage of mice reaching the criterion in the S trials across days represented as the cumulative probability. The Cre+ DCZ treated group needed additional days to reach criterion compared to controls. ( D ) Number of days for each mouse to reach the 70% success criterion in S trials (one-way ANOVA, F (2,31) = 3.77, p = 0.034; Tukey post hoc test indicates that the Cre+ DCZ group is significantly different from the Cre+ DMSO, while other pairs are not significantly different). ( E ) c-Fos+ cell density in all groups of mice. There was an increase in c-Fos+ mGCs in mice in which ≤7-week abDGCs were inhibited by DCZ ( H (2) = 7.22, p = 0.0271; Kruskal–Wallis) (Cre+ DCZ comparison to Cre+ DMSO, p = 0.014; Cre+ DCZ comparison to Cre- DCZ, p = 0.027 with Dunn’s multiple comparison post hoc analysis). ( F ) Example section after c-Fos immunohistochemistry to assess the distribution of c-Fos+ mGCs between the IB and SB of the dorsal DG in groups of mice who received DCZ (cre− vs cre+). c-Fos cells are localized closer to the subgranular zone (SGZ) in the Cre+ group. Calibration bar, 100 μm. ( G ) c-Fos+ density in the CA3 region of the hippocampus in each of the groups of mice. No significant difference was observed in any of the groups (one-way ANOVA, F (2,13) = 2.27, p = 0.143; Tukey post hoc test indicates that there is no significant difference between any pairs of groups). ( H ) Blade distribution of c-Fos+ mGCs in the IB and SB of the dorsal DG in mice performing the LS separation receiving either DCZ or DMSO 30 min prior to task performance. Active neurons are distributed preferentially to the SB than the IB in the DG for the control groups. However, this distribution bias is significantly reduced in the Cre+ DCZ group (two-way ANOVA (unbalanced) (blade: F (1,24) = 659.55, p < 2.2 × 10⁻¹⁶; group: F (2,24) = 3.48 × 10⁻¹³, p = 1; group × blade: F (1,16) = 0.040, p = 0.845). Tukey post hoc test. Significant differences were observed for IB comparisons: Cre+ DCZ vs Cre− DCZ (p = 0.000548) and Cre+ DCZ vs Cre+ DMSO (p = 0.00765). Significant differences were observed for SB comparisons: Cre+ DCZ vs Cre− DCZ (p = 0.000548) and Cre+ DCZ vs Cre+ DMSO (p = 0.00765)). ( I ) Spatial distribution of c-Fos+ GCs along hilar to molecular layer axes of the SB (0–120 μm) and the IB (0–120 μm) of the dorsal DG, with 0 μm indicating the hilar position. c-Fos+ cells are localized closer to the SGZ in the Cre+ DCZ group. ( J ) Cartoon illustrating the DG with its two blades: IB and SB. Dentate granule cells are depicted in gray, while red circles represent activity labeled cells in the DG. For Cre+ DCZ mice, the distribution of labeled neurons is closer to SGZ, away from outer radial granule cell layer (GCL) and more evenly distributed in IB and SB.

Journal: eLife

Article Title: Subregional activity in the dentate gyrus is amplified during elevated cognitive demands

doi: 10.7554/eLife.109611

Figure Lengend Snippet: ( A ) Schematic representation of the pattern separation paradigm. Prior to the onset of the training, mice (males n = 17, females n = 16) were provided tamoxifen-containing food ad libitum in the home cage for 7 weeks to activate TAM inducible Cre recombinase (Ascl1-CreER T2 ) in abDGCs. All mice received DCZ (deschloroclozapine, 50 μg kg −1 ) or DMSO injections 30 min before the beginning of LS trials. ( B ) Example sections from Ascl1-CreER T2 ; hM4Di mice with immunohistochemical localization of the HA tagged DREADD (top) and after LS training with c-Fos+ neurons labeled in red. Calibration bars: 140 μm. ( C ) Percentage of mice reaching the criterion in the S trials across days represented as the cumulative probability. The Cre+ DCZ treated group needed additional days to reach criterion compared to controls. ( D ) Number of days for each mouse to reach the 70% success criterion in S trials (one-way ANOVA, F (2,31) = 3.77, p = 0.034; Tukey post hoc test indicates that the Cre+ DCZ group is significantly different from the Cre+ DMSO, while other pairs are not significantly different). ( E ) c-Fos+ cell density in all groups of mice. There was an increase in c-Fos+ mGCs in mice in which ≤7-week abDGCs were inhibited by DCZ ( H (2) = 7.22, p = 0.0271; Kruskal–Wallis) (Cre+ DCZ comparison to Cre+ DMSO, p = 0.014; Cre+ DCZ comparison to Cre- DCZ, p = 0.027 with Dunn’s multiple comparison post hoc analysis). ( F ) Example section after c-Fos immunohistochemistry to assess the distribution of c-Fos+ mGCs between the IB and SB of the dorsal DG in groups of mice who received DCZ (cre− vs cre+). c-Fos cells are localized closer to the subgranular zone (SGZ) in the Cre+ group. Calibration bar, 100 μm. ( G ) c-Fos+ density in the CA3 region of the hippocampus in each of the groups of mice. No significant difference was observed in any of the groups (one-way ANOVA, F (2,13) = 2.27, p = 0.143; Tukey post hoc test indicates that there is no significant difference between any pairs of groups). ( H ) Blade distribution of c-Fos+ mGCs in the IB and SB of the dorsal DG in mice performing the LS separation receiving either DCZ or DMSO 30 min prior to task performance. Active neurons are distributed preferentially to the SB than the IB in the DG for the control groups. However, this distribution bias is significantly reduced in the Cre+ DCZ group (two-way ANOVA (unbalanced) (blade: F (1,24) = 659.55, p < 2.2 × 10⁻¹⁶; group: F (2,24) = 3.48 × 10⁻¹³, p = 1; group × blade: F (1,16) = 0.040, p = 0.845). Tukey post hoc test. Significant differences were observed for IB comparisons: Cre+ DCZ vs Cre− DCZ (p = 0.000548) and Cre+ DCZ vs Cre+ DMSO (p = 0.00765). Significant differences were observed for SB comparisons: Cre+ DCZ vs Cre− DCZ (p = 0.000548) and Cre+ DCZ vs Cre+ DMSO (p = 0.00765)). ( I ) Spatial distribution of c-Fos+ GCs along hilar to molecular layer axes of the SB (0–120 μm) and the IB (0–120 μm) of the dorsal DG, with 0 μm indicating the hilar position. c-Fos+ cells are localized closer to the SGZ in the Cre+ DCZ group. ( J ) Cartoon illustrating the DG with its two blades: IB and SB. Dentate granule cells are depicted in gray, while red circles represent activity labeled cells in the DG. For Cre+ DCZ mice, the distribution of labeled neurons is closer to SGZ, away from outer radial granule cell layer (GCL) and more evenly distributed in IB and SB.

Article Snippet: Strain, strain background ( Mus musculus ) , R26-LSL-Gi-DREADD (hM4Di) , Jackson Laboratory , JAX 026219 , Chemogenetic inhibition model.

Techniques: Immunohistochemical staining, Labeling, Comparison, Immunohistochemistry, Control, Activity Assay

The effects of chronic CM DRN modulation on htau behaviors. A, Graphical overview of the htau DREADDs experiment. Htau mice were intracranially administered hM3Dq ( n = 14), hM4Di ( n = 14), or control virus ( n = 9) at 2 months’ age. DREADDs agonist (CNO DHC) was administered via water bottle for 6 weeks. B, C, RNAscope assessment of Fos expression in virally transduced Tph2 + cells before CNO washout (acute) and after 7‐day washout. Two data points were collected per mouse in (C). No statistical test was performed in (C), data were qualitatively compared. D, E, NOR. D, Percent preference for novel object and (E), discrimination index. 1w ANOVA with Tukey post hoc. F, G, NSF. F, Latency to feed and (G), time spent in the corners of the arena during 10‐minute assessment period. H–K, EPM task. H, Percent of total time spent in open arm, (I) entries to open arm, (J) total distance traveled, and (K) average velocity during the task. L–R, Three‐chamber SIT. L, Total social interaction time, (M) number of social interactions, (N) social interaction time as a percentage of time spent in the social chamber, and (O) time spent in the center chamber. P–R, Representative heat maps, normalized within trial. D–F, H–K, 1w ANOVA with Dunnett post hoc. G, 1w Welch ANOVA with Dunnett post hoc. L–O, 1w ANOVA with Tukey post hoc. * p < 0.05, ** p < 0.01. 1w, one way; ANOVA, analysis of variance; CM, centromedial; CNO, clozapine‐n‐oxide; DHC, dihydrochloride; DREADDs, designer receptors exclusively activated by designer drugs; DRN, dorsal raphe nucleus; EPM, elevated plus maze; NOR, novel objection recognition; NSF, novelty‐induced suppression of feeding; SIT, social interaction test.

Journal: Alzheimer's & Dementia

Article Title: Selective reduction of KCNA4 in vulnerable glutamatergic–serotonin neurons of the dorsal raphe nucleus in Alzheimer's disease

doi: 10.1002/alz.71317

Figure Lengend Snippet: The effects of chronic CM DRN modulation on htau behaviors. A, Graphical overview of the htau DREADDs experiment. Htau mice were intracranially administered hM3Dq ( n = 14), hM4Di ( n = 14), or control virus ( n = 9) at 2 months’ age. DREADDs agonist (CNO DHC) was administered via water bottle for 6 weeks. B, C, RNAscope assessment of Fos expression in virally transduced Tph2 + cells before CNO washout (acute) and after 7‐day washout. Two data points were collected per mouse in (C). No statistical test was performed in (C), data were qualitatively compared. D, E, NOR. D, Percent preference for novel object and (E), discrimination index. 1w ANOVA with Tukey post hoc. F, G, NSF. F, Latency to feed and (G), time spent in the corners of the arena during 10‐minute assessment period. H–K, EPM task. H, Percent of total time spent in open arm, (I) entries to open arm, (J) total distance traveled, and (K) average velocity during the task. L–R, Three‐chamber SIT. L, Total social interaction time, (M) number of social interactions, (N) social interaction time as a percentage of time spent in the social chamber, and (O) time spent in the center chamber. P–R, Representative heat maps, normalized within trial. D–F, H–K, 1w ANOVA with Dunnett post hoc. G, 1w Welch ANOVA with Dunnett post hoc. L–O, 1w ANOVA with Tukey post hoc. * p < 0.05, ** p < 0.01. 1w, one way; ANOVA, analysis of variance; CM, centromedial; CNO, clozapine‐n‐oxide; DHC, dihydrochloride; DREADDs, designer receptors exclusively activated by designer drugs; DRN, dorsal raphe nucleus; EPM, elevated plus maze; NOR, novel objection recognition; NSF, novelty‐induced suppression of feeding; SIT, social interaction test.

Article Snippet: After a 7‐day recovery period, mice were continuously administered the DREADD agonist CNO DHC (clozapine‐n‐oxide dihydrochloride; cat. #HB6149; Hello Bio) at a concentration of 6.06 mg/mL (equivalent to 5 mg/mL CNO) via water bottle in a home‐cage environment.

Techniques: Control, Virus, RNAscope, Expressing

( a ) Scheme of the implantation and injection. ( b ) Coronal section of the cerebellum showing hM4Di-DREADD expression in the three CN. ( c ) Representative confocal image showing hM4Di-DREADD expression on neuronal membranes. ( d ) Electrode placement close to cells expressing hM4Di-DREADD (red: lesion site, yellow: electrode track). ( e ) Examples of high-pass filtered traces of CN recordings before and after CNO injection. ( f ) Examples of spike shapes obtained from spike sorting in CN (line + shading indicates mean +/- SD). ( g ) Boxplots displaying the percentage of modulation of CN neurons average firing rate induced by CNO or SAL injections during an open field session. CN firing rate was reduced after 1 mg/kg of CNO injection in DREADD-injected mice compared to other groups (Wilcoxon test* p<0.05, ***p<0.001. Boxes represent quartiles and whiskers correspond to range; points are singled as outliers if they deviate more than 1.5 x interquartile range from the nearest quartile). ( h ) Latency to fall during fixed speed rotarod (5, 10, 15, 20 r.p.m.) for all experimental groups. One way repeated-measure ANOVA on averaged values for all the speed steps in each experimental group followed by Tukey Post hoc pairwise comparison (p>0.05 in all cases). ( i ) Impact of daily injections of CNO before trial 1 on accelerating rotarod performance. Summary of the performance for each trial/day (repeated measure ANOVA Group effect *p<0.05, ***p<0.001; # p<0.05, ### Pp <0.001 Tukey pairwise test last trial of each day vs first trial of next day).( j ) Impact of daily injections of CNO after the task (30 min after trial 7). All treatment is switched to saline in the Late phase. Same presentation as in ( i ). Data in h,i,j are presented as mean ± S.E.M. n indicates the number of mice.

Journal: eLife

Article Title: Multiple functions of cerebello-thalamic neurons in learning and offline consolidation of a motor skill in mice

doi: 10.7554/eLife.102813

Figure Lengend Snippet: ( a ) Scheme of the implantation and injection. ( b ) Coronal section of the cerebellum showing hM4Di-DREADD expression in the three CN. ( c ) Representative confocal image showing hM4Di-DREADD expression on neuronal membranes. ( d ) Electrode placement close to cells expressing hM4Di-DREADD (red: lesion site, yellow: electrode track). ( e ) Examples of high-pass filtered traces of CN recordings before and after CNO injection. ( f ) Examples of spike shapes obtained from spike sorting in CN (line + shading indicates mean +/- SD). ( g ) Boxplots displaying the percentage of modulation of CN neurons average firing rate induced by CNO or SAL injections during an open field session. CN firing rate was reduced after 1 mg/kg of CNO injection in DREADD-injected mice compared to other groups (Wilcoxon test* p<0.05, ***p<0.001. Boxes represent quartiles and whiskers correspond to range; points are singled as outliers if they deviate more than 1.5 x interquartile range from the nearest quartile). ( h ) Latency to fall during fixed speed rotarod (5, 10, 15, 20 r.p.m.) for all experimental groups. One way repeated-measure ANOVA on averaged values for all the speed steps in each experimental group followed by Tukey Post hoc pairwise comparison (p>0.05 in all cases). ( i ) Impact of daily injections of CNO before trial 1 on accelerating rotarod performance. Summary of the performance for each trial/day (repeated measure ANOVA Group effect *p<0.05, ***p<0.001; # p<0.05, ### Pp <0.001 Tukey pairwise test last trial of each day vs first trial of next day).( j ) Impact of daily injections of CNO after the task (30 min after trial 7). All treatment is switched to saline in the Late phase. Same presentation as in ( i ). Data in h,i,j are presented as mean ± S.E.M. n indicates the number of mice.

Article Snippet: The DREADD ligand Clozapine-N-Oxide (CNO, TOCRIS, Bristol, UK) was dissolved in SAL (0.9% sodium chloride) and injected intraperitoneally at 1 mg/kg.

Techniques: Injection, Expressing, Comparison, Saline

DAPI-positive neurons expressing hM4Di-DREADD-mCherry in cerebellar nuclei.

Journal: eLife

Article Title: Multiple functions of cerebello-thalamic neurons in learning and offline consolidation of a motor skill in mice

doi: 10.7554/eLife.102813

Figure Lengend Snippet: DAPI-positive neurons expressing hM4Di-DREADD-mCherry in cerebellar nuclei.

Article Snippet: The DREADD ligand Clozapine-N-Oxide (CNO, TOCRIS, Bristol, UK) was dissolved in SAL (0.9% sodium chloride) and injected intraperitoneally at 1 mg/kg.

Techniques: Expressing

( a ) Footprint patterns were quantitatively assessed for three parameters as shown on representative footprint patterns for all experimental groups. Three parameters are represented graphically: linear movement (bottom left), sigma (bottom middle), and alternation coefficient (bottom right). ( b ) Tests of strength and coordination. Grid test: latency reflecting the time before falling from the grid. 30 s of cut-off was established as the maximum latency (dotted line on figure). Horizontal bar: latency to cross the horizontal bar (balance beam test) for all experimental groups. Vertical pole: latency to reach home cage in vertical pole test for all experimental groups. ( c ) Locomotor activity (Velocity) in DREADD and non-DREADD (Sham) injected mice after CNO or SAL injection during open-field sessions before (OF1) and after (OF2) rotarod for experimental days 1, 4, and 7. n indicates the number of mice. Boxes in a,b represent quartiles and whiskers correspond to range; points are singled as outliers if they deviate more than 1.5 x interquartile range from the nearest quartile. Data in c are represented as mean ± S.E.M. No post hoc significant difference was found for the data presented in this figure.

Journal: eLife

Article Title: Multiple functions of cerebello-thalamic neurons in learning and offline consolidation of a motor skill in mice

doi: 10.7554/eLife.102813

Figure Lengend Snippet: ( a ) Footprint patterns were quantitatively assessed for three parameters as shown on representative footprint patterns for all experimental groups. Three parameters are represented graphically: linear movement (bottom left), sigma (bottom middle), and alternation coefficient (bottom right). ( b ) Tests of strength and coordination. Grid test: latency reflecting the time before falling from the grid. 30 s of cut-off was established as the maximum latency (dotted line on figure). Horizontal bar: latency to cross the horizontal bar (balance beam test) for all experimental groups. Vertical pole: latency to reach home cage in vertical pole test for all experimental groups. ( c ) Locomotor activity (Velocity) in DREADD and non-DREADD (Sham) injected mice after CNO or SAL injection during open-field sessions before (OF1) and after (OF2) rotarod for experimental days 1, 4, and 7. n indicates the number of mice. Boxes in a,b represent quartiles and whiskers correspond to range; points are singled as outliers if they deviate more than 1.5 x interquartile range from the nearest quartile. Data in c are represented as mean ± S.E.M. No post hoc significant difference was found for the data presented in this figure.

Article Snippet: The DREADD ligand Clozapine-N-Oxide (CNO, TOCRIS, Bristol, UK) was dissolved in SAL (0.9% sodium chloride) and injected intraperitoneally at 1 mg/kg.

Techniques: Activity Assay, Injection

( a ) Example of evolution of latencies to fall for a mouse during the accelerating rotarod protocol. Linear regressions estimated values of trials 1 and 7 during each day are shown using green hollow dots, within-day learning (green arrows), overnight change (red arrows), and day + night learning (brown arrows). ( b ) Evolution of estimated daily learning, overnight change, and consolidated learning shows learning mostly in days 1–4 (Early Phase). Plots represent mean ± S.E.M. ( c ) Within-day learning vs daily initial performances (top) and within-day learning vs consolidated learning (bottom) in the Early phase. Scatterplot of performance from all control mice; the ellipse contains 50% of a bivariate normal distribution fitted to the values and the dot indicates the center of the distribution. Deming linear regression outcomes are represented with 95% confidence interval in shaded color. ( d ) Same as panel (c) with superimposition of controls (black) and mice expressing DREADD in DCN and CNO during the task (orange); only ellipses and regression line are included for clarity of the graph (**p<0.01 ***p<0.001 Wilcoxon test for difference between groups of residuals, i.e. signed distance of performances to Deming regression line of control mice). ( e ) Same as panel ( d ) for CNO administered after the task.

Journal: eLife

Article Title: Multiple functions of cerebello-thalamic neurons in learning and offline consolidation of a motor skill in mice

doi: 10.7554/eLife.102813

Figure Lengend Snippet: ( a ) Example of evolution of latencies to fall for a mouse during the accelerating rotarod protocol. Linear regressions estimated values of trials 1 and 7 during each day are shown using green hollow dots, within-day learning (green arrows), overnight change (red arrows), and day + night learning (brown arrows). ( b ) Evolution of estimated daily learning, overnight change, and consolidated learning shows learning mostly in days 1–4 (Early Phase). Plots represent mean ± S.E.M. ( c ) Within-day learning vs daily initial performances (top) and within-day learning vs consolidated learning (bottom) in the Early phase. Scatterplot of performance from all control mice; the ellipse contains 50% of a bivariate normal distribution fitted to the values and the dot indicates the center of the distribution. Deming linear regression outcomes are represented with 95% confidence interval in shaded color. ( d ) Same as panel (c) with superimposition of controls (black) and mice expressing DREADD in DCN and CNO during the task (orange); only ellipses and regression line are included for clarity of the graph (**p<0.01 ***p<0.001 Wilcoxon test for difference between groups of residuals, i.e. signed distance of performances to Deming regression line of control mice). ( e ) Same as panel ( d ) for CNO administered after the task.

Article Snippet: The DREADD ligand Clozapine-N-Oxide (CNO, TOCRIS, Bristol, UK) was dissolved in SAL (0.9% sodium chloride) and injected intraperitoneally at 1 mg/kg.

Techniques: Control, Expressing

( a ) Scheme of the combined viral injections targeting the CN->CL neurons using a retrograde virus expressing the Cre in the thalamus and a virus inducing Cre-dependent expression of inhibitory DREADD in the CN. Top : schematic of the viral injections. Bottom : GFP fluorescence revealing the site of injection of the CAV viruses. ( b ) Comparison of the effect of daily injections before trial 1 of CNO (orange) or Saline (black) in mice described in panel ( a ) (Data represents mean ± S.E.M, n indicates the number of mice; *p<0.05, **p<0.01, ***p<0.001; repeated measure ANOVA Group effect). ( c ) Same as ( b ) for daily CNO injections 30 min after trial 7 during the Early Phase. ( d ) Scheme of combined viral injections targeting the CN->VAL neurons. ( e,f ) Same as ( b,c ) for CNO-injected (blue) and control (black) mice obtained as described in panel d. ( g ) Same as for the Saline-treated groups of panels b–f. ( h–k ) Same as for the different groups receiving injections CNO during or after the task compared to Saline-treated mice. Same color coding as panels b, c and e, f. (**p<0.01 ***p<0.001 Wilcoxon test for difference between groups of residuals i.e. signed distance of performances to Deming regression line of control mice). CN: cerebellar nuclei, VAL: ventral anterior lateral thalamus, CL: central lateral thalamus.

Journal: eLife

Article Title: Multiple functions of cerebello-thalamic neurons in learning and offline consolidation of a motor skill in mice

doi: 10.7554/eLife.102813

Figure Lengend Snippet: ( a ) Scheme of the combined viral injections targeting the CN->CL neurons using a retrograde virus expressing the Cre in the thalamus and a virus inducing Cre-dependent expression of inhibitory DREADD in the CN. Top : schematic of the viral injections. Bottom : GFP fluorescence revealing the site of injection of the CAV viruses. ( b ) Comparison of the effect of daily injections before trial 1 of CNO (orange) or Saline (black) in mice described in panel ( a ) (Data represents mean ± S.E.M, n indicates the number of mice; *p<0.05, **p<0.01, ***p<0.001; repeated measure ANOVA Group effect). ( c ) Same as ( b ) for daily CNO injections 30 min after trial 7 during the Early Phase. ( d ) Scheme of combined viral injections targeting the CN->VAL neurons. ( e,f ) Same as ( b,c ) for CNO-injected (blue) and control (black) mice obtained as described in panel d. ( g ) Same as for the Saline-treated groups of panels b–f. ( h–k ) Same as for the different groups receiving injections CNO during or after the task compared to Saline-treated mice. Same color coding as panels b, c and e, f. (**p<0.01 ***p<0.001 Wilcoxon test for difference between groups of residuals i.e. signed distance of performances to Deming regression line of control mice). CN: cerebellar nuclei, VAL: ventral anterior lateral thalamus, CL: central lateral thalamus.

Article Snippet: The DREADD ligand Clozapine-N-Oxide (CNO, TOCRIS, Bristol, UK) was dissolved in SAL (0.9% sodium chloride) and injected intraperitoneally at 1 mg/kg.

Techniques: Virus, Expressing, Fluorescence, Injection, Comparison, Saline, Control

( a ) Footprint patterns were quantitatively assessed for three parameters as shown on representative footprint patterns (top) for all experimental groups. Three parameters are represented graphically: linear movement (bottom left), sigma (bottom middle), and alternation coefficient (bottom right). ( b ) Latency reflecting the time before falling from the grid. 30 seconds of cut-off was established as the maximum latency (dotted line on figure). ( c ) Latency to cross the horizontal bar (balance beam test) for all experimental groups. ( d ) Latency to reach home cage in vertical pole test for all experimental groups. ( e ) Locomotor activity (Velocity) in DREADD-injected mice after CNO or SAL injection during open-fields sessions before (OF1) and after (OF2) rotarod for days 1, 4, and 7 (**p<0.01 paired t-test OF1 vs OF2). ( f ) Latency to fall during fixed speed rotarod (5, 10, 15, 20 r.p.m.) for all experimental groups. One-way repeated measures ANOVA was performed on averaged values for each speed step in each experimental group followed by a Tukey post hoc pairwise comparison (*p<0.05, **p<0.01 for CN->VAL group CNO vs SAL). CN, cerebellar nuclei, CL, centrolateral thalamus; VAL, ventral anterior lateral thalamus. n indicates the number of mice. Boxes represent quartiles and whiskers correspond to range; points are singled out as outliers if they deviate more than 1.5 x interquartile range from the nearest quartile. Plots in e,f represent mean ± S.E.M.

Journal: eLife

Article Title: Multiple functions of cerebello-thalamic neurons in learning and offline consolidation of a motor skill in mice

doi: 10.7554/eLife.102813

Figure Lengend Snippet: ( a ) Footprint patterns were quantitatively assessed for three parameters as shown on representative footprint patterns (top) for all experimental groups. Three parameters are represented graphically: linear movement (bottom left), sigma (bottom middle), and alternation coefficient (bottom right). ( b ) Latency reflecting the time before falling from the grid. 30 seconds of cut-off was established as the maximum latency (dotted line on figure). ( c ) Latency to cross the horizontal bar (balance beam test) for all experimental groups. ( d ) Latency to reach home cage in vertical pole test for all experimental groups. ( e ) Locomotor activity (Velocity) in DREADD-injected mice after CNO or SAL injection during open-fields sessions before (OF1) and after (OF2) rotarod for days 1, 4, and 7 (**p<0.01 paired t-test OF1 vs OF2). ( f ) Latency to fall during fixed speed rotarod (5, 10, 15, 20 r.p.m.) for all experimental groups. One-way repeated measures ANOVA was performed on averaged values for each speed step in each experimental group followed by a Tukey post hoc pairwise comparison (*p<0.05, **p<0.01 for CN->VAL group CNO vs SAL). CN, cerebellar nuclei, CL, centrolateral thalamus; VAL, ventral anterior lateral thalamus. n indicates the number of mice. Boxes represent quartiles and whiskers correspond to range; points are singled out as outliers if they deviate more than 1.5 x interquartile range from the nearest quartile. Plots in e,f represent mean ± S.E.M.

Article Snippet: The DREADD ligand Clozapine-N-Oxide (CNO, TOCRIS, Bristol, UK) was dissolved in SAL (0.9% sodium chloride) and injected intraperitoneally at 1 mg/kg.

Techniques: Activity Assay, Injection, Comparison

( a ) Performance of mice with DREADD expression in CL-projecting CN neurons, which learned the task under respectively CNO and Saline treatment during 7 days and then received, respectively, Saline and CNO treatment during a Reversal phase (*: p<0.05 repeated-measure ANOVA group effect; Data represented as mean ± S.E.M, n indicates the number of mice). ( b ) Change induced by treatment switch on skill levels for the first trial (daily start) and last trial (daily end); values are estimated as in , and start and end values for days 8 and 9 are averaged for each animal; *p<0.05, **p<0.01, ***p<0.001 paired Wilcoxon test. Boxes represent quartiles and whiskers correspond to range; points are singled as outliers if they deviate more than 1.5 x interquartile range from the nearest quartile. ( c,d ) Same as panels a and b for mice with DREADD expression in VAL-projecting CN neurons . CN: cerebellar nuclei, VAL: ventral anterior lateral thalamus, CL: central lateral thalamus.

Journal: eLife

Article Title: Multiple functions of cerebello-thalamic neurons in learning and offline consolidation of a motor skill in mice

doi: 10.7554/eLife.102813

Figure Lengend Snippet: ( a ) Performance of mice with DREADD expression in CL-projecting CN neurons, which learned the task under respectively CNO and Saline treatment during 7 days and then received, respectively, Saline and CNO treatment during a Reversal phase (*: p<0.05 repeated-measure ANOVA group effect; Data represented as mean ± S.E.M, n indicates the number of mice). ( b ) Change induced by treatment switch on skill levels for the first trial (daily start) and last trial (daily end); values are estimated as in , and start and end values for days 8 and 9 are averaged for each animal; *p<0.05, **p<0.01, ***p<0.001 paired Wilcoxon test. Boxes represent quartiles and whiskers correspond to range; points are singled as outliers if they deviate more than 1.5 x interquartile range from the nearest quartile. ( c,d ) Same as panels a and b for mice with DREADD expression in VAL-projecting CN neurons . CN: cerebellar nuclei, VAL: ventral anterior lateral thalamus, CL: central lateral thalamus.

Article Snippet: The DREADD ligand Clozapine-N-Oxide (CNO, TOCRIS, Bristol, UK) was dissolved in SAL (0.9% sodium chloride) and injected intraperitoneally at 1 mg/kg.

Techniques: Expressing, Saline

M1-WT-HA (n=11-14) (A) , M1-KO (n=13-16) (B) or M1-PD-HA (n=11-14) (C) mice were treated with 1.5 mg/kg scopolamine +/-10 mg/kg VU846 30 mins prior to training in a fear conditioning paradigm. Mice were then returned to the same environment 24 hrs later for contextual memory retrieval whereby a reduced level of freezing indicates impaired memory. The response in vehicle treated animals across strains was also compared (D) . 2-way ANOVA with Tukey’s post-hoc correction for multiple comparisons, *p<0.05, ***p<0.001, ****p<0.0001, ns=not significant. (E ) Representative images of CA1 hippocampal region of HA-tagged M1-WT, M1-DREADD, M1-PD and M1-DREADD-PD stained with HA antibody demonstrating location of the receptor (green). Images at 40X magnification, scale bar = 50 µm

Journal: bioRxiv

Article Title: The importance of M1 muscarinic receptor phosphorylation in learning and memory

doi: 10.64898/2026.03.23.713145

Figure Lengend Snippet: M1-WT-HA (n=11-14) (A) , M1-KO (n=13-16) (B) or M1-PD-HA (n=11-14) (C) mice were treated with 1.5 mg/kg scopolamine +/-10 mg/kg VU846 30 mins prior to training in a fear conditioning paradigm. Mice were then returned to the same environment 24 hrs later for contextual memory retrieval whereby a reduced level of freezing indicates impaired memory. The response in vehicle treated animals across strains was also compared (D) . 2-way ANOVA with Tukey’s post-hoc correction for multiple comparisons, *p<0.05, ***p<0.001, ****p<0.0001, ns=not significant. (E ) Representative images of CA1 hippocampal region of HA-tagged M1-WT, M1-DREADD, M1-PD and M1-DREADD-PD stained with HA antibody demonstrating location of the receptor (green). Images at 40X magnification, scale bar = 50 µm

Article Snippet: The drugs utilised in this study were the pan-muscarinic antagonist, to induce a learning and memory (LM) deficit, scopolamine hydrobromide (Sigma) at 0.5-3 mg/kg doses; the M1 positive allosteric modulator (PAM) VU0486846 (VU846) at 10-100 mg/kg; and the M1-DREADD activating ligand clozapine-n-oxide (CNO) (Tocris) at 0.3 mg/kg.

Techniques: Staining

(A) Male and female M1-WT-HA (n=11) and M1-PD (n=12) mice were treated with the M1-PAM VU846 30 mins prior to fear conditioning and returned to the chamber 24 hrs later for contextual memory retrieval. 2-way ANOVA ****p<0.0001 effect of strain, no significant effect of drug overall p=0.12, Bonferroni’s post-hoc correction for multiple comparisons between vehicle and VU846 for each strain, ns= not significant. (B & C) Male M1-DREADD mice were treated with 0.3 mg/kg of clozapine-N-oxide (CNO), an orthosteric agonist of the DREADD receptor, or vehicle (n=4-5 per group) 30 mins prior to training in a fear conditioning paradigm and then returned to the same environment 24 hrs later for contextual memory retrieval (B) or presentation of the tone in an altered environment 48 hrs later for cued memory retrieval (C) whereby a reduced level of freezing indicates impaired memory. (D) WT mice (n=7-9 per group) were also treated with 0.3 mg/kg CNO in a separate experiment demonstrating no effect of CNO. 2-way ANOVA with Dunnett’s post-hoc correction for multiple comparisons, ****p<0.0001, all other comparisons p>0.05.

Journal: bioRxiv

Article Title: The importance of M1 muscarinic receptor phosphorylation in learning and memory

doi: 10.64898/2026.03.23.713145

Figure Lengend Snippet: (A) Male and female M1-WT-HA (n=11) and M1-PD (n=12) mice were treated with the M1-PAM VU846 30 mins prior to fear conditioning and returned to the chamber 24 hrs later for contextual memory retrieval. 2-way ANOVA ****p<0.0001 effect of strain, no significant effect of drug overall p=0.12, Bonferroni’s post-hoc correction for multiple comparisons between vehicle and VU846 for each strain, ns= not significant. (B & C) Male M1-DREADD mice were treated with 0.3 mg/kg of clozapine-N-oxide (CNO), an orthosteric agonist of the DREADD receptor, or vehicle (n=4-5 per group) 30 mins prior to training in a fear conditioning paradigm and then returned to the same environment 24 hrs later for contextual memory retrieval (B) or presentation of the tone in an altered environment 48 hrs later for cued memory retrieval (C) whereby a reduced level of freezing indicates impaired memory. (D) WT mice (n=7-9 per group) were also treated with 0.3 mg/kg CNO in a separate experiment demonstrating no effect of CNO. 2-way ANOVA with Dunnett’s post-hoc correction for multiple comparisons, ****p<0.0001, all other comparisons p>0.05.

Techniques:

Male M1-DREADD mice were treated with 0.01 - 0.3 mg/kg of clozapine-N-oxide (CNO), an orthosteric agonist of the DREADD receptor, or vehicle (n=5-9 per group) 30 mins prior to training in a fear conditioning paradigm and then returned to the same environment 24 hrs later for contextual memory retrieval. Data presented as freezing level normalised to the highest freezing response. One way ANOVA with Bonferroni’s post hoc correction for multiple comparisons to vehicle treatment group *p<0.05. All other comparisons not significant.

Journal: bioRxiv

Article Title: The importance of M1 muscarinic receptor phosphorylation in learning and memory

doi: 10.64898/2026.03.23.713145

Figure Lengend Snippet: Male M1-DREADD mice were treated with 0.01 - 0.3 mg/kg of clozapine-N-oxide (CNO), an orthosteric agonist of the DREADD receptor, or vehicle (n=5-9 per group) 30 mins prior to training in a fear conditioning paradigm and then returned to the same environment 24 hrs later for contextual memory retrieval. Data presented as freezing level normalised to the highest freezing response. One way ANOVA with Bonferroni’s post hoc correction for multiple comparisons to vehicle treatment group *p<0.05. All other comparisons not significant.

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