slc17a7 cre mice  (Addgene inc)

Journal: Molecular Therapy Advances

Article Title: Adeno-associated virus-induced neurotoxicity is prevented by CpG depletion

doi: 10.1016/j.omta.2026.201703

Figure Lengend Snippet: CpG depletion does not alter transgene expression but prevents AAV-induced dendritic loss (A) Immunoblot of S1 homogenates probed with antibodies against transgene product EGFP and loading control GAPDH following injection of 1 × 10 10 vg of unmodified AAV1-pCAG-FLEX-EGFP or CpG-depleted AAV1-pCAG-FLEX-EGFP into Slc17a7-IRES2-Cre-D mice. Each lane contains samples from a different animal. UT, untreated. (B) Densitometry shows that EGFP protein levels are indistinguishable in ipsilateral S1 of Slc17a7-IRES2-Cre-D mice injected with either CpG-depleted or unmodified virus. EGFP protein levels are normalized to GAPDH protein levels in the same lane and represented as a fold change from the AAV1-treated condition. (C) Representative cortical neurons from untreated wild-type mice or ipsilateral (injected) hemispheres of wild-type mice injected with 1 × 10 10 vg of either unmodified or CpG-depleted AAV1-pCAG-FLEX-EGFP. Scale bars, 100 μm. (D) Total dendritic length and (E) total Sholl intersections of cortical neurons from untreated wild-type mice and wild-type mice injected with 1 × 10 10 vg of either unmodified or CpG-depleted AAV1-pCAG-FLEX-EGFP. Values are represented as a ratio of injected (ipsi) normalized to uninjected (contra) hemispheres. (F–H) Sholl intersections plotted as a function of distance from the soma for cortical neurons from the same animals shown in (E). (D and E), n = 4 untreated control mice, 10–14 neurons per animal (average = 12 cells); n = 5 mice injected with unmodified AAV1-pCAG-FLEX-EGFP, 10–14 neurons per animal (average = 12 cells); n = 5 mice injected with CpG-depleted AAV1-pCAG-FLEX-EGFP, 12–15 neurons per animal (average = 13 cells). Data in (B and D–H) are represented as mean ± SD. ∗∗∗∗ p < .0001; ∗∗∗ p < .001; ∗∗ p < .01; ns, not significant. In (G), a single ∗ indicates p < 0.05, and vertically stacked ∗ indicate p < 0.01. Statistical analysis: in (B, F–H), two-tailed unpaired t test; in (D) and (E), one-way ANOVA with Tukey’s multiple comparison test.

Article Snippet: Male C57BL/6J and Slc17a7-IRES2-Cre-D (Strain # 037512) mice were obtained from Jackson Laboratory (The Jackson Laboratory, Bar Harbor, ME).

Techniques: Expressing, Western Blot, Control, Injection, Virus, Two Tailed Test, Comparison

Journal: Nature

Article Title: A combinatorial neural code for long-term motor memory

doi: 10.1038/s41586-024-08193-3

Figure Lengend Snippet: a . Top left, representative video frame with automatically labeled tongue markers using DeepLabCut. Top middle, superimposed tongue tip trajectories and x and y velocities of individual lick events during lick left (red) and lick right (blue). Data from an example mouse across sessions within the same task context. Tongue tip trajectory scale bar, 4 pixels (x) and 6 pixels (y). X velocity scale bar, 12 ms and 2 pixels/s. Y velocity scale bar, 12 ms and 1.5 pixels/s. Top right, scatter of averaged pairwise similarity of single lick events (Pearson’s correlation) calculated within session versus across sessions. Data from two mice. Bottom, same as top but for data across task contexts 1 and 2. b . Same as a , but for jaw marker analysis. Jaw tip trajectory scale bar, 4 pixels (x) and 4 pixels (y). X velocity scale bar, 12 ms and 1 pixels/s. Y velocity scale bar, 12 ms and 1.5 pixels/s. c . Left, schematics of learning speed under two models. Context-specific saving effect (top): faster re-learning only for previously learned tasks. Context non-specific saving effect (bottom): faster learning each time. Right, faster reversal learning is consistent with a context-specific saving effect. Re-learning of task context 2’ is significantly faster than initial learning of task context 2 (top). P = 0.0487, paired t-test. Circles indicate individual mice (N = 13 mice). Crosses indicate mean ± s.e.m. We examine task context 2 because the initial learning of task context 1 is confounded by the exposure to home-cage training. To examine context non-specific saving effect, we compare the speed of re-learning task context 1’ versus re-learning task context 2’ (bottom). The two conditions have similar task-specific prior training. No significant difference is observed. P = 0.3425, two-tailed paired t-test. d . Same as Fig. , but separately plotting photoinhibition results for task context 1 (left) and task context 2 (right). e . Experimental timeline of an example mouse imaged within the same task context over extended time. Black, behavior training in automated home-cage. Gray, habituation in two-photon setup. Red, calcium imaging in two-photon setup. All the trials are concatenated. Black triangle indicates the end of learning voluntary head-fixation and start of learning in tactile instructed licking task. Averaging window, 100 trials. f . Same as e , but for two mice imaged across different task contexts. g . Summary plot of experimental timeline from all GP4.3 mice used for imaging in this study. h-i . Behavior performance curves for the initial learning from GP4.3 mice ( h , n = 15 mice, all were trained in automated home-cage) and Slc17a7-Cre x Ai148 mice ( i , n = 11 mice, 7 mice were trained in automated home-cage and 4 mice were manually trained). Different colors represent individual mice. Circles indicate end of the learning curves for GP4.3 mice and termination of training for Slc17a7-Cre x Ai148 mice. j . Behavior performance within imaging sessions across 4 segments of trials. Thin gray lines indicate individual sessions. Thick black lines indicate mean ± s.e.m. Data from Fig. .

Article Snippet: Eleven Slc17a7-Cre mice (JAX 023527) crossed to Cre-dependent GCaMP6f reporter Ai148 mice (JAX 030328) were used for behaviour training but were not used for calcium imaging due to poor behavioural performance (Extended Data Fig. ).

Techniques: Labeling, Marker, Two Tailed Test, Imaging

Journal: eLife

Article Title: Vglut2-based glutamatergic signaling in central noradrenergic neurons is dispensable for normal breathing and chemosensory reflexes

doi: 10.7554/eLife.88673

Figure Lengend Snippet: ( A ) Breeding schematic to generate the three intersectional reporter lines Slc17a7 Cre ; Dbh p2a-Flpo ; Rosa26 RC::FLTG , Slc17a6 Cre ; Dbh p2a-Flpo ; Rosa26 RC::FLTG , Slc17a8 Cre ; Dbh p2a-Flpo ; Rosa26 RC::FLTG . In each of the three intersectional reporter crosses, NA neurons co-expressing either Vglut1, Vglut2, or Vglut3 are labeled by green fluorescent protein (eGFP) while NA neurons without any Vglut1, 2, or 3 expressions are labeled by red fluorescent protein (tdTomato). Shown as an example is the Slc17a6 Cre ; Dbh p2a-Flpo ; Rosa26 RC::FLTG (Vglut2-Cre/+; DBH-p2a-Flpo/+; RC::FLTG/+) intersectional reporter line. ( B ) Quantification of the percentage of Vglut2-coexpressing NA neurons among NA neurons in each brainstem NA nuclei including A7, LC, A5, sub CD/CV, anterior C1/A1 and C2/A2, posterior C1/A1 and C2/A2. Pink data points represent female data while blue data points represent male data. ( C ) Quantification of the percentage of Vglut3-coexpressing NA neurons among NA neurons in each NA nucleus. Pink data points represent female data while blue data points represent male data. ( D ) Fluorescent expression of tdTomato (red) and eGFP (green) in coronal sections of Vglut1, Vglut2, or Vglut3 intersectional reporter lines in brainstem NA nuclei. Nucleus is labeled by 4′,6-diamidino-2-phenylindole (DAPI) (blue). Scale bar 50 μm. Figure 1—source data 1. Cell quantification for Vglut2-coexpressing noradrenergic (NA) neurons in the cumulative fate map. Figure 1—source data 2. Cell quantification for Vglut3-coexpressing noradrenergic (NA) neurons in the cumulative fate map.

Article Snippet: To verify Vglut2 co-expression in LC in adult mice , 6-week-old Slc17a7 Cre mice were injected with pAAV-EF1a-DIO-tdTomato-WPRE virus (RRID: Addgene_133786 , obtained from Joshua Ortiz at the Optogenetics and Viral Vectors Core at the Jan and Dan Duncan Neurological Research Institute, 1 ul at 7.90E+10 Gc/ml) into the LC (coordinates from bregma anteroposterior −5.4 mm, lateral +0.8 mm, and dorsoventral −4.0 mm) and allowed to incubate for 4 weeks.

Techniques: Expressing, Labeling

Journal: eLife

Article Title: Vglut2-based glutamatergic signaling in central noradrenergic neurons is dispensable for normal breathing and chemosensory reflexes

doi: 10.7554/eLife.88673

Figure Lengend Snippet: ( A ) Representative images of Slc17a7 (Vglut1), Slc17a6 (Vglut2), and Slc17a8 (Vglut3) and Dbh (DBH) double ISH in coronal sections of WT mice in all of brainstem NA nuclei in adult mice. DBH (green), Vglut1/2/3 (red), DAPI (cyan). Scale bar 50 μm. ( B ) Quantification of the percentage of Slc17a6 (Vglut2) co-expression in NA neurons in each NA nucleus. Female data (pink), male data (blue). ( C ) Quantification of the percentage of Slc17a8 (Vglut3) co-expression in NA neurons in each NA nucleus. Female data (pink), male data (blue). Figure 2—source data 1. Cell quantification for Vglut2-coexpressing noradrenergic (NA) neurons in the in situ hybridization. Figure 2—source data 2. Cell quantification for Vglut3-coexpressing noradrenergic (NA) neurons in the in situ hybridization.

Article Snippet: To verify Vglut2 co-expression in LC in adult mice , 6-week-old Slc17a7 Cre mice were injected with pAAV-EF1a-DIO-tdTomato-WPRE virus (RRID: Addgene_133786 , obtained from Joshua Ortiz at the Optogenetics and Viral Vectors Core at the Jan and Dan Duncan Neurological Research Institute, 1 ul at 7.90E+10 Gc/ml) into the LC (coordinates from bregma anteroposterior −5.4 mm, lateral +0.8 mm, and dorsoventral −4.0 mm) and allowed to incubate for 4 weeks.

Techniques: Expressing, In Situ Hybridization

Journal: eLife

Article Title: Vglut2-based glutamatergic signaling in central noradrenergic neurons is dispensable for normal breathing and chemosensory reflexes

doi: 10.7554/eLife.88673

Figure Lengend Snippet: ( A ) Representative images of Slc17a7 (Vglut1), Slc17a6 (Vglut2), and Slc17a8 (Vglut3) and Dbh (DBH) double ISH in coronal sections of WT mice in C3 NA nuclei in adult mice. DBH (green), Vglut1/2/3 (red), DAPI (cyan). Scale bar 50 μm. ( B ) Quantification of the percentage of Slc17a6 (Vglut2)/ Slc17a8 (Vglut3) co-expression in C3 NA nucleus. Female data (pink), male data (blue).

Article Snippet: To verify Vglut2 co-expression in LC in adult mice , 6-week-old Slc17a7 Cre mice were injected with pAAV-EF1a-DIO-tdTomato-WPRE virus (RRID: Addgene_133786 , obtained from Joshua Ortiz at the Optogenetics and Viral Vectors Core at the Jan and Dan Duncan Neurological Research Institute, 1 ul at 7.90E+10 Gc/ml) into the LC (coordinates from bregma anteroposterior −5.4 mm, lateral +0.8 mm, and dorsoventral −4.0 mm) and allowed to incubate for 4 weeks.

Techniques: Expressing