aav5-gfaabc1d-cyto-gcamp6f  (Addgene inc)

Journal: bioRxiv

Article Title: Cell-autonomous mitochondrial calcium flux governs oligodendrocyte regeneration

doi: 10.1101/2025.11.18.689087

Figure Lengend Snippet: ( a ) Experimental timeline. Mice expressing GCaMP6f in oligodendroglia were implanted with a gradient index (GRIN) lens above the corpus callosum of the M1 motor cortex, ∼1.8 mm below the brain surface, and exposed to the CPZ diet for 6 weeks. UCLA Miniscope V4 baseplating was performed in the 5th week of CPZ demyelination and microendoscopy imaging was performed in weeks 5 and 6 of demyelination (orange) as well as during weeks 1 to 6 of remyelination (gray). ( b ) Miniscope recordings were performed in an open field and locomotion behavior was monitored. ( c,d ) Representative projection images of output by Occam with designated active ROIs (white lines) detected in several substacks as obtained with the Miniscope configuration of Occam (File S1) . Two substack projections obtain from image stacks are shown per week ( c ). Ca 2+ activity traces ( c , bottom) and quantification ( d ) of in vivo oligodendroglial Ca 2+ imaging during de-and re-myelination (not significant; one-way repeated measures ANOVA). ( e ) Open field locomotion maps during in vivo Ca 2+ imaging, recorded under demyelination and remyelination conditions in the same mouse. ( f ) Correlations between Ca 2+ signals and distance traveled, speed and motor activity duration (not significant Pearson correlations) during remyelination. Data represented as mean ± s.e.m. Scale bars 100 µm ( b ) and 20 µm ( c ).

Article Snippet: Adult Pdgfrα CreERT( ± ); Gcamp5-tdTomato Lox/Lox and Pdgfrα CreERT( ± ) ;Gcamp6f Lox/Lox mice were intraperitoneally injected with tamoxifen (MP Biomedicals Germany GmbH, CAS: 10540-29-1) to induce the expression of GCaMP5 or GCaMP6f in OPCs and their progeny, respectively.

Techniques: Expressing, Imaging, Activity Assay, In Vivo

Journal: bioRxiv

Article Title: Cell-autonomous mitochondrial calcium flux governs oligodendrocyte regeneration

doi: 10.1101/2025.11.18.689087

Figure Lengend Snippet: ( a ) Experimental timeline. Callosal LPC-induced lesions were induced in control mice (Pdgfrɑ CreER(-/-) ;hM3D(Gq)-YFP lox/+ ;GCaMP6f lox/lox ) and experimental mice (Pdgfrɑ CreER(+/-) ;hM3D(Gq)-YFP lox/+ ;GCaMP6f lox/lox mice) which express both GCaMP6f and the hM3D(Gq) receptor in oligodendroglial cells (Inset: OLIG2 + YFP + cells). Mice were either used to prepare acute slices at 7 dpi or received i.p. CNO injections (1 mg/kg) at 7, 9, 11 and 13 dpi before perfusion at 14 dpi. ( b ) Representative projection image output by Occam with designated active ROIs (white lines) and Ca 2+ traces (middle) from a lesion at 7 dpi. CNO bath application significantly increased Ca 2+ activity compared to baseline. Dot plots (right) showing significant Ca 2+ activity increases in the presence of CNO in lesions (p-value from Wilcoxon matched-pairs signed rank test). ( c ) Representative image of immunofluorescence staining for MBP (magenta), OLIG2 (green) and CC1 (white) in a callosal lesion (dashed line). Insets from white rectangle shows an OLIG2 + CC1 - OPC (open arrow) and mature OLIG2 + CC1 + OL (closed arrow). ( d-f ) Densities of OLIG2 + lineage cells ( d ), OLIG2 + CC1 - OPCs ( e ) and OLIG2 + CC1 + OLs ( f ) in Cre -/- (control) and Cre +/- mice following CNO injections. ( g ) Percentages of OLIG2 + cells that are OPCs (OLIG2 + CC1 - ) or mature OLs (OLIG2 + CC1 + ) did not differ between CNO-treated mice with and without oligodendroglial hM3D(Gq) receptor expression. p-values from mixed linear models with type II likelihood ratio tests ( d-g ). Data is presented as mean ± s.e.m. with open circles representing slices and closed circles averages per mouse ( d,f ). Not significant n.s. Scale bars are 5 µm in ( a ), 10 µm in ( b ) and 50 µm and 10 µm in ( c ).

Article Snippet: Adult Pdgfrα CreERT( ± ); Gcamp5-tdTomato Lox/Lox and Pdgfrα CreERT( ± ) ;Gcamp6f Lox/Lox mice were intraperitoneally injected with tamoxifen (MP Biomedicals Germany GmbH, CAS: 10540-29-1) to induce the expression of GCaMP5 or GCaMP6f in OPCs and their progeny, respectively.

Techniques: Control, Activity Assay, Immunofluorescence, Staining, Expressing

Journal: bioRxiv

Article Title: Complementary cortical and thalamic contributions to cell-type-specific striatal activity dynamics during movement

doi: 10.1101/2025.07.11.664473

Figure Lengend Snippet: A) Schematic of the motor task. B) Left, schematic of the approach used for labeling dMSNs and iMSNs in DLS. Bottom, timeline of experiments. Right, schematic of the approach used for labeling the presynaptic cortical and thalamic inputs to dMSNs and iMSNs. C) Schematic of the imaged brain regions and maximum intensity projection images from 2-photon in vivo imaging, showing neurons that express GCaMP6f in DLS, M1, M2, and PF. Scale bar: 20µm. D) Left, trial-averaged activity heatmaps of neurons in DLS, ordered based on the maximum peak time of their activity. Middle, population average activity of all DLS neurons. Right, population average activity of dMSNs (blue) and iMSNs (orange), mean ± SEM. n = 337 dMSNs (from N = 21 mice) and n = 414 iMSNs (from N = 12 mice). E) Same as D) for the input neurons in M1, M2 and PF. M1: n = 180 dMSN-projecting neurons (from N = 5 mice) and 249 iMSN-projecting neurons (from N = 4 mice); M2: n = 1849 dMSN-projecting neurons (from N =12 mice) and 2962 iMSN-projecting neurons (from N = 12 mice); PF: n = 136 dMSN-projecting neurons (from N = 8 mice) and 150 iMSN-projecting neurons (from N = 9 mice). DLS: dorsolateral striatum; M1: primary motor cortex; M2: secondary motor cortex; PF: parafascicular nucleus of the thalamus.

Article Snippet: For 2-photon imaging of dMSNs and iMSNs in the dorsolateral striatum, AAV1-hSyn-FLEX-GCaMP6f (Addgene, Catalog # 100833-AAV1) was injected via beveled glass pipettes into the right hemisphere of Drd1-Cre and Adora2a-Cre mice respectively, at coordinates: 0.5 mm anterior and 2.2 mm lateral from bregma, at depths 2.2, 2.3 and 2.4 mm from the pia (∼200 nL at each depth, at a rate of 20 nL per minute.

Techniques: Labeling, In Vivo Imaging, Activity Assay