inhibitory dreadd virus addgene plasmid  (Addgene inc)

Journal: Neuron

Article Title: Spatial and Temporal Locomotor Learning in Mouse Cerebellum.

doi: 10.1016/j.neuron.2019.01.038

Figure Lengend Snippet: Figure 6. Differential Lateralization of Spatial and Temporal Learning in the Cerebellum (A) Experimental protocol for assessing lateralization of split-belt learning. Animals with unilateral cerebellar injections underwent adaptation protocols where the fast belt was either ipsilateral (ipsi-fast, right) or contralateral (contra-fast, left) to the injection. (B) Cerebellar section from a mouse with a unilateral injection of retrograde DREADDs in the interposed. The borders of the interposed nucleus (ip) are indicated by a white, dashed line. Sample was stained with DAPI (blue) and anti-mCherry (for DREADD expression). (C) Average front-limb center of oscillation asymmetry (spatial) adaptation curves for ipsi-fast (left, pink) and contra-fast (right, purple) conditions. In each plot, black curves represent mice under saline conditions; colors represent CNO. The split-belt period is indicated by a gray, shaded patch. (D) Average center of oscillation aftereffects (first 2 washout trials minus average baseline) for ipsi- and contra-fast conditions with saline and CNO. Individual animals are shown with thin, dashed lines. (E) Average double support for ipsi- and contra- ast conditions, plotted as in (C). (F) Average center of oscillation aftereffects for ipsi- and contra-fast conditions with saline and CNO. Significance was assessed with paired t tests. Errors bars represent SEM.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Bacterial and Virus Strains Retrograde inhibitory DREADD virus Addgene plasmid #44362 Inhibitory DREADD virus Addgene plasmid #50475 Experimental Models: Organisms/Strains Mouse: Purkinje cell degeneration Jackson Labs 0537 Mouse: Reeler Jackson Labs 000235 Mouse: L7-cre Jackson Labs 004146 Mouse: C57BL6J Jackson Labs 000664 Software and Algorithms MATLAB (versions 2014b and 2015b) Mathworks http://www.mathworks.com LabView 2012 National Instruments http://ni.com R 3.5.1 The R Project for Statistical Computing http://www.r-project.org/ LocoMouse Tracker Carey lab http://www.github.com/careylab/ LocoMouse Other Treadmill motors Maxon DCX22S GB KL 12V; Planetary gearhead - GPX22 LN 35:1; Sensor - ENX16 EASY Motor controllers Maxon ESCON 50/5, 4-Q Servocontroller for DC/ EC motors, 5/15 A, 10 - 50 VDC Firewire camera Allied Vision PIKE F-032 B Data acquisition board: camera framegrabber National Instruments NI PCIe-8253, IEEE 1394b Board with Vision Acquisition SW Data acquisition board National Instruments NI PCIe-6323, X Series

Techniques: Injection, Staining, Expressing, Saline

Journal: Neuron

Article Title: Spatial and Temporal Locomotor Learning in Mouse Cerebellum.

doi: 10.1016/j.neuron.2019.01.038

Figure Lengend Snippet: Figure 4. Locomotor Adaptation Requires Projections to the Interposed Cerebellar Nu- cleus (A) Cerebellar circuit diagram depicting retrograde approach for targeting inhibitory DREADDs to Pur- kinje cells terminating in each of the three cerebellar nuclei. Cerebellar structures and cell types are abbreviated as follows: dcn: deep cerebellar nuclei; mf: mossy fibers; pf: parallel fibers; PC: Purkinje cells. (B) Mean virus expression area for medial, inter- posed, and lateral nucleus injections, overlaid on mouse brain atlas. Colorbar represents percent of mice with expression in a particular zone. (C–E) Mean step length adaptation curves for medial (C, blue; n = 7 mice), interposed (D, pink; n = 10 mice), and lateral (E, green; n = 6 mice) nucleus injections, following administration of saline (black) or CNO (colors). Gray-shaded patch indicates split- belt trials. (F–H) Average step-length aftereffect for saline and CNO conditions for medial (F), interposed (G), and lateral (H) nuclei. Individual animals are represented with thin, dashed lines and open circles; averages are represented with closed symbols. Significance was assessed with paired t tests. Errors bars represent SEM. See also Figure S5 and Figure S6.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Bacterial and Virus Strains Retrograde inhibitory DREADD virus Addgene plasmid #44362 Inhibitory DREADD virus Addgene plasmid #50475 Experimental Models: Organisms/Strains Mouse: Purkinje cell degeneration Jackson Labs 0537 Mouse: Reeler Jackson Labs 000235 Mouse: L7-cre Jackson Labs 004146 Mouse: C57BL6J Jackson Labs 000664 Software and Algorithms MATLAB (versions 2014b and 2015b) Mathworks http://www.mathworks.com LabView 2012 National Instruments http://ni.com R 3.5.1 The R Project for Statistical Computing http://www.r-project.org/ LocoMouse Tracker Carey lab http://www.github.com/careylab/ LocoMouse Other Treadmill motors Maxon DCX22S GB KL 12V; Planetary gearhead - GPX22 LN 35:1; Sensor - ENX16 EASY Motor controllers Maxon ESCON 50/5, 4-Q Servocontroller for DC/ EC motors, 5/15 A, 10 - 50 VDC Firewire camera Allied Vision PIKE F-032 B Data acquisition board: camera framegrabber National Instruments NI PCIe-8253, IEEE 1394b Board with Vision Acquisition SW Data acquisition board National Instruments NI PCIe-6323, X Series

Techniques: Virus, Expressing, Saline

Journal: Neuron

Article Title: Spatial and Temporal Locomotor Learning in Mouse Cerebellum.

doi: 10.1016/j.neuron.2019.01.038

Figure Lengend Snippet: Figure 6. Differential Lateralization of Spatial and Temporal Learning in the Cerebellum (A) Experimental protocol for assessing lateralization of split-belt learning. Animals with unilateral cerebellar injections underwent adaptation protocols where the fast belt was either ipsilateral (ipsi-fast, right) or contralateral (contra-fast, left) to the injection. (B) Cerebellar section from a mouse with a unilateral injection of retrograde DREADDs in the interposed. The borders of the interposed nucleus (ip) are indicated by a white, dashed line. Sample was stained with DAPI (blue) and anti-mCherry (for DREADD expression). (C) Average front-limb center of oscillation asymmetry (spatial) adaptation curves for ipsi-fast (left, pink) and contra-fast (right, purple) conditions. In each plot, black curves represent mice under saline conditions; colors represent CNO. The split-belt period is indicated by a gray, shaded patch. (D) Average center of oscillation aftereffects (first 2 washout trials minus average baseline) for ipsi- and contra-fast conditions with saline and CNO. Individual animals are shown with thin, dashed lines. (E) Average double support for ipsi- and contra- ast conditions, plotted as in (C). (F) Average center of oscillation aftereffects for ipsi- and contra-fast conditions with saline and CNO. Significance was assessed with paired t tests. Errors bars represent SEM.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Bacterial and Virus Strains Retrograde inhibitory DREADD virus Addgene plasmid #44362 Inhibitory DREADD virus Addgene plasmid #50475 Experimental Models: Organisms/Strains Mouse: Purkinje cell degeneration Jackson Labs 0537 Mouse: Reeler Jackson Labs 000235 Mouse: L7-cre Jackson Labs 004146 Mouse: C57BL6J Jackson Labs 000664 Software and Algorithms MATLAB (versions 2014b and 2015b) Mathworks http://www.mathworks.com LabView 2012 National Instruments http://ni.com R 3.5.1 The R Project for Statistical Computing http://www.r-project.org/ LocoMouse Tracker Carey lab http://www.github.com/careylab/ LocoMouse Other Treadmill motors Maxon DCX22S GB KL 12V; Planetary gearhead - GPX22 LN 35:1; Sensor - ENX16 EASY Motor controllers Maxon ESCON 50/5, 4-Q Servocontroller for DC/ EC motors, 5/15 A, 10 - 50 VDC Firewire camera Allied Vision PIKE F-032 B Data acquisition board: camera framegrabber National Instruments NI PCIe-8253, IEEE 1394b Board with Vision Acquisition SW Data acquisition board National Instruments NI PCIe-6323, X Series

Techniques: Injection, Staining, Expressing, Saline