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(A) Experimental scheme showing rectal AKP transplantation into Lepr-Cre(+);Chrm3 fl/fl and control mice. IVIS, in vivo imaging. (B and C) Mouse survival (B) and tumor-derived luciferase signals (C). n=6 ( Lepr-Cre(−);Chrm3 fl/fl ) and 9 mice ( Lepr-Cre(+);Chrm3 fl/fl ). (D) Non-phosphorylated YAP/PDGFRA co-IF. Red arrowheads; non-phosphorylated (active) YAP in PDGFRA + CAFs. cKO, conditional KO. n=5 mice. (E) NTN1/PDGFRA co-IF. n=5 mice. (F) VAChT IHC. Red arrowheads; VAChT + nerves. n=5 mice. (G) Experimental scheme showing rectal AKP transplantation with daily intratumor injection of the hM3Dq agonist <t>C21</t> (0.03 mg/kg) from Day 8. (H and I) Mouse survival (H) and tumor-derived luciferase signals (I). n=8 ( Lepr-Cre;LSL-hM3Dq(−);Ntn1 WT ), 7 ( Lepr-Cre;LSL-hM3Dq(+);Ntn1 WT ), 7 ( Lepr-Cre;LSL-hM3Dq(−);Ntn1 fl/fl ), and 7 mice ( Lepr-Cre;LSL-hM3Dq(+);Ntn1 fl/fl ). (J) Non-phosphorylated YAP/PDGFRA co-IF. Red/purple arrowheads; non-phosphorylated YAP in PDGFRA + CAFs in the Lepr-Cre;LSL-hM3Dq(+);Ntn1 WT / Lepr-Cre;LSL-hM3Dq(+);Ntn1 fl/fl groups. n=4 mice. (K) NTN1/PDGFRA co-IF. n=4 mice. (L) VAChT IHC. Green/red arrowheads:VAChT + nerves in the Lepr-Cre;LSL-hM3Dq(−);Ntn1 WT / Lepr-Cre;LSL-hM3Dq(+);Ntn1 WT groups. n=4 mice. Scale bars, 100 μm (A and G), 50/5 μm (main/insets; D, E, J, and K), 50 μm (F and L). Log-rank test (B and H), two-way repeated-measures ANOVA with post-hoc Sidak’s multiple comparison test at week 2 (C and I), two-tailed unpaired Student’s t-test (D-F), and two-way ANOVA followed by Tukey’s post-hoc multiple comparison tests (J-L). Mean±s.e.m. (C-F and I-L). See also Figures S5 .

C21, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 94/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Cholinergic signaling via muscarinic receptor 3 promotes colorectal cancer progression directly and indirectly through stroma-derived Netrin-1"

Article Title: Cholinergic signaling via muscarinic receptor 3 promotes colorectal cancer progression directly and indirectly through stroma-derived Netrin-1

Journal: Cancer cell

doi: 10.1016/j.ccell.2026.06.018

Figure Legend Snippet: (A) Experimental scheme showing rectal AKP transplantation into Lepr-Cre(+);Chrm3 fl/fl and control mice. IVIS, in vivo imaging. (B and C) Mouse survival (B) and tumor-derived luciferase signals (C). n=6 ( Lepr-Cre(−);Chrm3 fl/fl ) and 9 mice ( Lepr-Cre(+);Chrm3 fl/fl ). (D) Non-phosphorylated YAP/PDGFRA co-IF. Red arrowheads; non-phosphorylated (active) YAP in PDGFRA + CAFs. cKO, conditional KO. n=5 mice. (E) NTN1/PDGFRA co-IF. n=5 mice. (F) VAChT IHC. Red arrowheads; VAChT + nerves. n=5 mice. (G) Experimental scheme showing rectal AKP transplantation with daily intratumor injection of the hM3Dq agonist C21 (0.03 mg/kg) from Day 8. (H and I) Mouse survival (H) and tumor-derived luciferase signals (I). n=8 ( Lepr-Cre;LSL-hM3Dq(−);Ntn1 WT ), 7 ( Lepr-Cre;LSL-hM3Dq(+);Ntn1 WT ), 7 ( Lepr-Cre;LSL-hM3Dq(−);Ntn1 fl/fl ), and 7 mice ( Lepr-Cre;LSL-hM3Dq(+);Ntn1 fl/fl ). (J) Non-phosphorylated YAP/PDGFRA co-IF. Red/purple arrowheads; non-phosphorylated YAP in PDGFRA + CAFs in the Lepr-Cre;LSL-hM3Dq(+);Ntn1 WT / Lepr-Cre;LSL-hM3Dq(+);Ntn1 fl/fl groups. n=4 mice. (K) NTN1/PDGFRA co-IF. n=4 mice. (L) VAChT IHC. Green/red arrowheads:VAChT + nerves in the Lepr-Cre;LSL-hM3Dq(−);Ntn1 WT / Lepr-Cre;LSL-hM3Dq(+);Ntn1 WT groups. n=4 mice. Scale bars, 100 μm (A and G), 50/5 μm (main/insets; D, E, J, and K), 50 μm (F and L). Log-rank test (B and H), two-way repeated-measures ANOVA with post-hoc Sidak’s multiple comparison test at week 2 (C and I), two-tailed unpaired Student’s t-test (D-F), and two-way ANOVA followed by Tukey’s post-hoc multiple comparison tests (J-L). Mean±s.e.m. (C-F and I-L). See also Figures S5 .

Techniques Used: Expressing, Transplantation Assay, Control, In Vivo Imaging, Derivative Assay, Luciferase, Injection, Comparison, Two Tailed Test, Immunohistochemistry

(A) Luciferase (luc)-expressing AKP tumor organoids were co-treated with ACh and an AChR antagonist for 72 h. AKP tumor-derived luc signals were measured as a surrogate for viable cell numbers. Drug concentration as in Figure 2A . n=4. (B) Luc signals from control gRNA/g Chrm3 -expressing luc + AKP organoids treated (72 h). Bethanechol (Beth), 10 μM. n=4. (C) Gene set enrichment analysis of the YAP gene signature 53 using bulk RNA-seq from Beth-/vehicle-treated AKP organoids. The normalized enrichment score (NES) and false discovery rate (FDR)-adjusted P value are shown. n=3. (D) Western blotting of control gRNA/g Chrm3 -expressing AKP tumor cells treated (20 min). Red dotted rectangles: dephosphorylation of LATS1/YAP by ACh/Beth treatment. FR900359 (FR, Gq inhibitor), 1 μM. (E) Luc signals from luc + AKP organoids treated (72 h). VP, verteporfin (YAP inhibitor, 0.5 μM). n=5. (F) Experimental scheme showing rectal AKP transplantation with daily intratumor injection of the hM3Dq agonist C21 (0.03 mg/kg) from Day 8. (G and H) Mouse survival (G) and tumor-derived luc signals (H). n=10 ( Chat-Cre(−);hM3Dq mice; control AKP), 9 ( Chat-Cre(+);hM3Dq mice; control AKP), 9 ( Chat-Cre(−);hM3Dq mice; g Chrm3 AKP), and 9 mice ( Chat-Cre(+);hM3Dq mice; g Chrm3 AKP). (I) Non-phosphorylated YAP/luc (a marker for luc + AKP cancer cells) co-IF. Red arrowheads; non-phosphorylated YAP in luc + cancer cells. n=5 mice. Scale bars, 100 μm (A and F) and 50/5 μm (main/insets; I). One-way ANOVA followed by Tukey’s post-hoc multiple comparison tests (A, B, and E), gene set permutation tests (C), log-rank test (G), two-way repeated-measures ANOVA with post-hoc Sidak’s multiple comparison test at week 2 (H), and two-way ANOVA followed by Tukey’s post-hoc multiple comparison tests (I). Mean±s.e.m. (A, B, E, H, and I). See also Figures S6 and S7 ; Tables S3 and S4 .

Figure Legend Snippet: (A) Luciferase (luc)-expressing AKP tumor organoids were co-treated with ACh and an AChR antagonist for 72 h. AKP tumor-derived luc signals were measured as a surrogate for viable cell numbers. Drug concentration as in Figure 2A . n=4. (B) Luc signals from control gRNA/g Chrm3 -expressing luc + AKP organoids treated (72 h). Bethanechol (Beth), 10 μM. n=4. (C) Gene set enrichment analysis of the YAP gene signature 53 using bulk RNA-seq from Beth-/vehicle-treated AKP organoids. The normalized enrichment score (NES) and false discovery rate (FDR)-adjusted P value are shown. n=3. (D) Western blotting of control gRNA/g Chrm3 -expressing AKP tumor cells treated (20 min). Red dotted rectangles: dephosphorylation of LATS1/YAP by ACh/Beth treatment. FR900359 (FR, Gq inhibitor), 1 μM. (E) Luc signals from luc + AKP organoids treated (72 h). VP, verteporfin (YAP inhibitor, 0.5 μM). n=5. (F) Experimental scheme showing rectal AKP transplantation with daily intratumor injection of the hM3Dq agonist C21 (0.03 mg/kg) from Day 8. (G and H) Mouse survival (G) and tumor-derived luc signals (H). n=10 ( Chat-Cre(−);hM3Dq mice; control AKP), 9 ( Chat-Cre(+);hM3Dq mice; control AKP), 9 ( Chat-Cre(−);hM3Dq mice; g Chrm3 AKP), and 9 mice ( Chat-Cre(+);hM3Dq mice; g Chrm3 AKP). (I) Non-phosphorylated YAP/luc (a marker for luc + AKP cancer cells) co-IF. Red arrowheads; non-phosphorylated YAP in luc + cancer cells. n=5 mice. Scale bars, 100 μm (A and F) and 50/5 μm (main/insets; I). One-way ANOVA followed by Tukey’s post-hoc multiple comparison tests (A, B, and E), gene set permutation tests (C), log-rank test (G), two-way repeated-measures ANOVA with post-hoc Sidak’s multiple comparison test at week 2 (H), and two-way ANOVA followed by Tukey’s post-hoc multiple comparison tests (I). Mean±s.e.m. (A, B, E, H, and I). See also Figures S6 and S7 ; Tables S3 and S4 .

Techniques Used: Activation Assay, Luciferase, Expressing, Organoids, Derivative Assay, Concentration Assay, Control, Analysis, RNA Sequencing, Western Blot, De-Phosphorylation Assay, Transplantation Assay, Injection, Marker, Comparison

Activity Assay:

Article Title: Intralaminar thalamus relays basal ganglia output to the insular cortex to drive tic generation.
Article Snippet: .. To chemogenetically suppress the activity of specific neurons, the DREADD agonist clozapine-N-oxide (CNO; cat# HY-17366, MedChemExpress, Monmouth Junction, NJ, USA) was administered i.p. to mice expressing hM4Di-mCherry or mCherry (control) in IC neurons or IC-projecting dmTh neurons. .. CNO was dissolved in dimethyl sulfoxide (DMSO; cat# 13445-74, Nacalai Tesque, Kyoto, Japan) at 100 mg/mL to prepare stock aliquots, which were stored at − 30◦C.

Article Title: Thalamo-insular pathway regulates tic generation via motor-limbic crosstalk
Article Snippet: .. To chemogenetically suppress the activity of specific neurons, the DREADD agonist, clozapine-N-oxide (CNO; cat# HY-17366, MedChemExpress, Monmouth Junction, NJ, USA), was administered intraperitonially in mice expressing hM4Di-mCherry or mCherry (control) in IC neurons or IC-projecting dmTh neurons. .. CNO was dissolved in dimethyl sulfoxide (DMSO; cat# 13445-74, Nacalai Tesque, Kyoto, Japan) at 100 mg/ml to prepare stock aliquots, which were stored at –30°C.

Expressing:

Control:

Saline:

Article Title: Optogenetic activation of liver-innervating vagal sensory neurons increases anxiety-like behavior in mice
Article Snippet: To selectively inhibit LC NE neurons, a total volume of 200 nl of AAV8-EF1α-DIO-hM4D(Gi)-GFP (Applied Biological Materials, Inc, titer; 1.0 × 10 12 pfu/ml) was bilaterally injected into the LC (AP: −5.4 mm, DV: −3.0 mm, ML: ±0.8 mm). .. DREADD agonist 21 (MedChemExpress, 1 mg/kg) or saline was intraperitoneally administered 1 h prior to measurement of behavior test. .. The wireless optogenetic system (Neurolux) consists of a power distribution control box, three autotuners, three modular boxes, peripheral implantable optogenetic devices with μ-LED of 470 nm, and a laptop with NeuroLux software [ ].

Concentration Assay:

Article Title: Orbitofrontal noradrenaline acts as an early gate for reversal learning.
Article Snippet: .. The DREADD agonist deschloroclozapine (DCZ, MedChemExpress) was dissolved in dimethyl sulfoxide (DMSO) to a stock concentration of 50 mg/mL and stored at − 80◦C. ..

Image Search Results

Journal: Cancer cell

Article Title: Cholinergic signaling via muscarinic receptor 3 promotes colorectal cancer progression directly and indirectly through stroma-derived Netrin-1

doi: 10.1016/j.ccell.2026.06.018

Figure Lengend Snippet: (A) Experimental scheme showing rectal AKP transplantation into Lepr-Cre(+);Chrm3 fl/fl and control mice. IVIS, in vivo imaging. (B and C) Mouse survival (B) and tumor-derived luciferase signals (C). n=6 ( Lepr-Cre(−);Chrm3 fl/fl ) and 9 mice ( Lepr-Cre(+);Chrm3 fl/fl ). (D) Non-phosphorylated YAP/PDGFRA co-IF. Red arrowheads; non-phosphorylated (active) YAP in PDGFRA + CAFs. cKO, conditional KO. n=5 mice. (E) NTN1/PDGFRA co-IF. n=5 mice. (F) VAChT IHC. Red arrowheads; VAChT + nerves. n=5 mice. (G) Experimental scheme showing rectal AKP transplantation with daily intratumor injection of the hM3Dq agonist C21 (0.03 mg/kg) from Day 8. (H and I) Mouse survival (H) and tumor-derived luciferase signals (I). n=8 ( Lepr-Cre;LSL-hM3Dq(−);Ntn1 WT ), 7 ( Lepr-Cre;LSL-hM3Dq(+);Ntn1 WT ), 7 ( Lepr-Cre;LSL-hM3Dq(−);Ntn1 fl/fl ), and 7 mice ( Lepr-Cre;LSL-hM3Dq(+);Ntn1 fl/fl ). (J) Non-phosphorylated YAP/PDGFRA co-IF. Red/purple arrowheads; non-phosphorylated YAP in PDGFRA + CAFs in the Lepr-Cre;LSL-hM3Dq(+);Ntn1 WT / Lepr-Cre;LSL-hM3Dq(+);Ntn1 fl/fl groups. n=4 mice. (K) NTN1/PDGFRA co-IF. n=4 mice. (L) VAChT IHC. Green/red arrowheads:VAChT + nerves in the Lepr-Cre;LSL-hM3Dq(−);Ntn1 WT / Lepr-Cre;LSL-hM3Dq(+);Ntn1 WT groups. n=4 mice. Scale bars, 100 μm (A and G), 50/5 μm (main/insets; D, E, J, and K), 50 μm (F and L). Log-rank test (B and H), two-way repeated-measures ANOVA with post-hoc Sidak’s multiple comparison test at week 2 (C and I), two-tailed unpaired Student’s t-test (D-F), and two-way ANOVA followed by Tukey’s post-hoc multiple comparison tests (J-L). Mean±s.e.m. (C-F and I-L). See also Figures S5 .

Article Snippet: A Hamilton syringe holding a 33-gauge needle was used to inject C21 (0.03 mg/kg; HY-100234A, MedChemExpress; a volume of 10 μL, dissolved in DPBS) into the rectal tumor percutaneously through the anal skin.

Techniques: Expressing, Transplantation Assay, Control, In Vivo Imaging, Derivative Assay, Luciferase, Injection, Comparison, Two Tailed Test, Immunohistochemistry

Journal: Cancer cell

Article Title: Cholinergic signaling via muscarinic receptor 3 promotes colorectal cancer progression directly and indirectly through stroma-derived Netrin-1

doi: 10.1016/j.ccell.2026.06.018

Figure Lengend Snippet: (A) Luciferase (luc)-expressing AKP tumor organoids were co-treated with ACh and an AChR antagonist for 72 h. AKP tumor-derived luc signals were measured as a surrogate for viable cell numbers. Drug concentration as in Figure 2A . n=4. (B) Luc signals from control gRNA/g Chrm3 -expressing luc + AKP organoids treated (72 h). Bethanechol (Beth), 10 μM. n=4. (C) Gene set enrichment analysis of the YAP gene signature 53 using bulk RNA-seq from Beth-/vehicle-treated AKP organoids. The normalized enrichment score (NES) and false discovery rate (FDR)-adjusted P value are shown. n=3. (D) Western blotting of control gRNA/g Chrm3 -expressing AKP tumor cells treated (20 min). Red dotted rectangles: dephosphorylation of LATS1/YAP by ACh/Beth treatment. FR900359 (FR, Gq inhibitor), 1 μM. (E) Luc signals from luc + AKP organoids treated (72 h). VP, verteporfin (YAP inhibitor, 0.5 μM). n=5. (F) Experimental scheme showing rectal AKP transplantation with daily intratumor injection of the hM3Dq agonist C21 (0.03 mg/kg) from Day 8. (G and H) Mouse survival (G) and tumor-derived luc signals (H). n=10 ( Chat-Cre(−);hM3Dq mice; control AKP), 9 ( Chat-Cre(+);hM3Dq mice; control AKP), 9 ( Chat-Cre(−);hM3Dq mice; g Chrm3 AKP), and 9 mice ( Chat-Cre(+);hM3Dq mice; g Chrm3 AKP). (I) Non-phosphorylated YAP/luc (a marker for luc + AKP cancer cells) co-IF. Red arrowheads; non-phosphorylated YAP in luc + cancer cells. n=5 mice. Scale bars, 100 μm (A and F) and 50/5 μm (main/insets; I). One-way ANOVA followed by Tukey’s post-hoc multiple comparison tests (A, B, and E), gene set permutation tests (C), log-rank test (G), two-way repeated-measures ANOVA with post-hoc Sidak’s multiple comparison test at week 2 (H), and two-way ANOVA followed by Tukey’s post-hoc multiple comparison tests (I). Mean±s.e.m. (A, B, E, H, and I). See also Figures S6 and S7 ; Tables S3 and S4 .

Techniques: Activation Assay, Luciferase, Expressing, Organoids, Derivative Assay, Concentration Assay, Control, Analysis, RNA Sequencing, Western Blot, De-Phosphorylation Assay, Transplantation Assay, Injection, Marker, Comparison

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.

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

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.

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

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.

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

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.

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

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.

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

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