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.

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: 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.

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: 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.

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: 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.

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, Control, Injection, Battery, Virus

Journal: iScience

Article Title: Horizontal and vertical gene transfer shape the plasmid host range in surface-associated microbial systems

doi: 10.1016/j.isci.2026.115299

Figure Lengend Snippet: Hypothesis and experiment system (A) We hypothesize that vertical and horizontal gene transfer (VGT and HGT) are influenced by the characteristics of the potential recipient cell types and determine the proliferation and diversity of transconjugant cells. Because the potential recipient community comprises multiple cell types with varying growth traits and conjugation probabilities, we expect the resulting composition of transconjugant cells to be shaped by these cell type-specific traits. (B) Our experimental system consists of E . coli MG1655 lacI q -pLpp-mCherry as the plasmid donor strain and pB10 as the focal plasmid. pB10 donor cells express RFP from the chromosome and transconjugants express GFP from pB10.

Article Snippet: MBP- mCherry expression plasmid (Amp R ) , Addgene , Plasmid# 29747.

Techniques: Conjugation Assay, Plasmid Preparation

Journal: iScience

Article Title: Horizontal and vertical gene transfer shape the plasmid host range in surface-associated microbial systems

doi: 10.1016/j.isci.2026.115299

Figure Lengend Snippet: Transconjugant proportions and diversities after surface-associated conjugation assays for different environmental conditions (A) Proportion of transconjugant cells relative to total cells after surface-associated conjugation assays using the WWTP community as the potential recipient cell population. We conducted conjugation assays on 1×SWW, 10×SWW, or LB agar plates using E . coli MG1655 lacI q -pLpp-mCherry as the pB10 donor strain. (B) Relative abundances of bacterial class in the total potential recipient cell population (T) and the transconjugant cell population (TC) as identified by 16S rRNA gene sequencing. We separated and identified TC cells using FC-FACS-sorting of GFP-positive cells. (C) Normalized Shannon index of the transconjugant populations after surface-associated conjugation assays on 1×SWW, 10×SWW, or LB agar plates. We normalized the Shannon index of the TC populations to their corresponding T populations. (D) Principal coordinate analysis (PCoA) based on weighted UniFrac distances of T and TC populations after surface-associated conjugation assays on 1×SWW, 10×SWW, or LB agar plates. (E) Phylogenetic tree of transconjugant ASVs detected after surface-associated conjugation assays on 1×SWW, 10×SWW, or LB agar plates. The outer colored box denotes the bacterial phylum of each ASV, corresponding to the phylum-level groupings shown in panel (B). The inner heatmap box aligned with each tip shows the log 10 fold-changes in ASV abundance (TC relative to T) across the three conditions. For (A and C), each point is an independent biological replicate ( n = 3), horizontal bars are the means, error bars are ±1 standard deviation, and asterisks indicate statistically significant differences between the means based on two-way ANOVA with Holm correction (∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, ns = not significant). For (D), each point is an independent biological replicate ( n = 3).

Article Snippet: MBP- mCherry expression plasmid (Amp R ) , Addgene , Plasmid# 29747.

Techniques: Conjugation Assay, Sequencing, Standard Deviation

Journal: iScience

Article Title: Horizontal and vertical gene transfer shape the plasmid host range in surface-associated microbial systems

doi: 10.1016/j.isci.2026.115299

Figure Lengend Snippet: Transconjugant growth during surface-associated conjugation assays for different environmental conditions (A) Representative fluorescence microscopy images of transconjugant cells during surface-associated conjugation assays on LB agar plates. E . coli MG1655 lacI q -pLpp-mCherry is the pB10 donor strain and show red fluorescence. Transconjugant cells are green. The time indicated in the images refers to the point at which transconjugant cells first became detectable. (B) Normalized microcolony area ( A / a 0 ) plotted as a function of time during the surface-associated conjugation assays on LB agar plates. A is the total microcolony area and a 0 is the initial transconjugant area. Connected data points are for individual colonies ( n = 12). (C) Microcolony area at the endpoint of the mating assay (t = 24 h) for different environmental conditions. The half-violin and scatterplots present the sample distribution and individual microcolony measurements for surface-associated conjugation assays on different medium (n 1xSWW = 880, n 10xSWW = 664, n LB = 1,070, for microcolony number). We performed each experiment at least three independent experiments. Horizontal bars are the mean microcolony areas, error bars are the 99% confidence intervals, and asterisks indicate statistically significant differences between the means based on two-way ANOVA with Holm correction (∗∗ p < 0.01, ∗∗∗∗ p < 0.0001, ns = not significant).

Article Snippet: MBP- mCherry expression plasmid (Amp R ) , Addgene , Plasmid# 29747.

Techniques: Conjugation Assay, Fluorescence, Microscopy