mouse anti-mcherry primary antibody  (Beyotime)

Journal: bioRxiv

Article Title: Ventral pallidum GABA neurons bidirectionally control opioid relapse across rat behavioral models

doi: 10.1101/2022.02.03.479042

Figure Lengend Snippet: A) Expression of hM4Di-mCherry (red) localized largely within VP borders defined by substance P (green). B) Coronal sections depicting the center of hM4Di-mCherry expression (red) for each rat along VP’s rostrocaudal axis relative to bregma (substance P-defined VP borders = green). C) Expression of hM3Dq-mCherry (red) is similarly localized within VP borders (green). D) Coronal sections similarly depicting the center of hM3Dq-mCherry expression (red) for each rat is shown. E) CNO treatment in hM3Dq-mCherry rats tested in the homecage exhibited greater Fos in mCherry+ neurons (2 nd bar, blue), than in homecage mCherry-only rats treated with CNO (1 st bar, gray). CNO-treated hM3Dq-mCherry rats exposed to remifentanil-paired cues (3 rd bar, blue) or no cues (4 th bar, blue) had more Fos+ mCherry neurons homecage rats (1 st bar, gray). However, the hM3Dq stimulation of Fos was no different in the presence or absence of cues. Images above/embedded within bars depict 10x images of immunohistochemical staining of mCherry (brown) within VP borders, and Fos+ nuclei (black). Example mCherry-only and mCherry+Fos double-labeled neuron indicated with brown and brown/black arrows, respectively. Individual rat data shown as gray dots on top of bars. One-way ANOVA, Sidak post hoc: p *** ≤ 0.001. Data presented as mean ± SEM.

Article Snippet: Procedures mirrored above, except after the Fos stain, a mouse anti-mCherry primary antibody (Takara Bio, 1:5000) was used instead of the substance P primary antibody to visualize hM3Dq-mCherry or mCherry-expressing cells.

Techniques: Expressing, Immunohistochemical staining, Staining, Labeling

Journal: Neuropsychopharmacology

Article Title: The medial septum enhances reversal learning via opposing actions on ventral tegmental area and substantia nigra dopamine neurons

doi: 10.1038/s41386-019-0453-1

Figure Lengend Snippet: Proposed pathway from MS to midbrain and MS DREADD virus spread and expression. a Medial septum (MS) activation was shown previously to increase DA neuron population activity in the ventral tegmental area (VTA) and decrease it in the substantia nigra pars compacta (SNc) [23, 24]. Both effects required activation of the ventral subiculum (vSub), as intra-vSub infusions of TTX prevented both the VTA increase and the SNc decrease in DA neuron population activity [23]. The vSub activates the nucleus accumbens (NAc [26, 30]), which inhibits the ventral pallidum (VP [27]). VP regulation of midbrain DA neuron population activity was shown to be functionally divided along the rostral/caudal axis, with the rostral VP (rVP) selectively modulating DA neuron population activity changes in the VTA and the caudal VP (cVP) selectively modulating DA neuron population activity changes in the SNc [23]. VP inhibition disinhibits the VTA, leading to an increase in DA neuron population activity. In contrast, VP inhibition more potently disinhibits the substantia nigra pars reticulata (SNr) due to the greater sensitivity of GABAA receptors on reticulata GABAergic interneurons compared to SNc DA neurons [58–60]. Thus, DA neurons in the SNc are inhibited by SNr disinhibition, leading to a decrease in DA neuron population activity. b Diagram and corresponding representative photomicrograph showing the extent of the viral spread at the point of infusion, i.e., the area with the greatest expression and medial–lateral spread. This was well-contained within the MS and occurred at AP 0.48 for the majority of DREADD (red; right) and control (green; left) rats. Arrows denote the actual placement of the representative photomicrograph compared to the diagram to provide further context for overall viral placement in brain. Representative images at 30x magnification to show DREADD expression around cell bodies in MS. The c red staining is the endogenous reporter molecule, mCherry, and the d green is the mouse mCherry primary and goat anti-mouse (alexa 488) secondary stain. e The overlay shows co-staining of the endogenous reporter and the primary/secondary antibody stain, suggesting good expression of the DREADD receptor in the MS and good selectivity of the antibody for mCherry

Article Snippet: Slides were either stained with cresyl violet for histological verification of cannula placement [ 37 ] or stained with a mouse mCherry primary (1:8000, Abcam-ab125096) and goat anti-mouse secondary stain (alexa 488, 1:500; Abcam-ab150113) for verification of DREADD virus transfection and spread.

Techniques: Expressing, Activation Assay, Activity Assay, Inhibition, Staining

Journal: Neuropsychopharmacology

Article Title: The medial septum enhances reversal learning via opposing actions on ventral tegmental area and substantia nigra dopamine neurons

doi: 10.1038/s41386-019-0453-1

Figure Lengend Snippet: DREADD terminal expression and cannulae placements in vSub. Representative photomicrograph showing extensive DREADD terminal expression in the vSub at a 10x and b 20x magnification. Terminals (green) are stained with the mouse mCherry primary and goat anti-mouse secondary stain (alexa 488) and are shown near DAPI-stained cell bodies in vSub (blue) c Placement map showing the termination of the cannula location in vSub (blue dot) for each rat included in the reversal learning analysis. All rats included in the primary behavioral and electrophysiological analyses had placements within the vSub, while rats with cannula placements outside the vSub were analyzed separately (Fig. S.5). d Representative cannula placement showing termination of the cannula in the vSub. The top arrow marks the bottom of the guide cannula, and the bottom arrow marks the bottom of the infusion cannula

Article Snippet: Slides were either stained with cresyl violet for histological verification of cannula placement [ 37 ] or stained with a mouse mCherry primary (1:8000, Abcam-ab125096) and goat anti-mouse secondary stain (alexa 488, 1:500; Abcam-ab150113) for verification of DREADD virus transfection and spread.

Techniques: Expressing, Staining