dta coding sequence  (Addgene inc)

Journal: bioRxiv

Article Title: Oligodendrocyte dependent synchronized activity orchestrates circuit maturation and brain functionalization

doi: 10.1101/2024.05.06.590880

Figure Lengend Snippet: a, Diagram illustrating the injection of AAV-hMAG-mCherry and AAV-hMAG-DTA into the mouse cerebellum at early postnatal days (P6-7). b, Fluorescence microscopy image showing mCherry expression (red) in the section of cerebellum at P14 following AAV-hMAG-mCherry injection at P7. Scale bar, 300 µm. c, Specificity of mCherry expression (red) in ASPA-positive oligodendrocytes (green) at P14 by AAV-hMAG-mCherry injection at P7. Scale bar, 100 µm. d, Scatter plot graph depicting the percentage of RFP and ASPA double-positive cells among RFP-positive populations (from 2 mice). e, Sequential visualization of oligodendrocyte deletion over time, indicated by ASPA staining in cerebellar sections from control (AAV-hMAG-mCherry) and DTA-treated (AAV-hMAG-DTA) mice at P10, P14, P21, and P78. Scale bar, 100 µm. f, Quantification of ASPA-positive cell density at each stage (from 3 mice per group per each stage). Bars and dots indicate mean and data from individual fields of view, respectively. g-i, Decrease in correlation coefficients (CCs) indicating reduced synchrony of spontaneous activities among PC population at P13-15 following DTA-mediated oligodendrocyte ablation. g, Representative time-course of spontaneous calcium transients by extracting relative fluorescence changes by in vivo calcium imaging captured from 25 regions of interest (ROIs) in the cerebellum of control and DTA-expressing mice at P14. Scale bar = 20 s. Y-axis is ΔF/F0 = (F - F0)/(F0). h, Correlation coefficient matrices for spontaneous calcium transient activity across multiple ROIs in CTL (upper panel) and DTA (lower panel) mice, with the color scale indicating the strength of the correlation between pairs of ROIs. i, Scatter plot graph of CCs for ROI pairs, categorized by separation distances of 0–80 mm and 120–200 mm (analyzed from four mice per group). Lines and plots indicate mean and data from individual separation distances, respectively. j-l, Quantification of the frequency, amplitude, and integrated area of calcium transients in control (AAV-hMAG-mCherry) and DTA-treated (AAV-hMAG-DTA) mice at P13-15 (data from four mice per group). Bars and dots indicate mean and data from individual ROIs, respectively. **** p < 0.0001 (Mann-Whitney U test).

Article Snippet: For AAV-hMAG-DTA, we amplified the DTA coding sequence from the pAAV-mCherry-flex-dtA (a gift from Naoshige Uchida, Addgene plasmid # 58536) using primer pairs (Table.

Techniques: Injection, Fluorescence, Microscopy, Expressing, Staining, Control, In Vivo, Imaging, Activity Assay, MANN-WHITNEY

Journal: bioRxiv

Article Title: Oligodendrocyte dependent synchronized activity orchestrates circuit maturation and brain functionalization

doi: 10.1101/2024.05.06.590880

Figure Lengend Snippet: a , Representative traces of CF-EPSCs in PCs from control (CTL) and AAV-hMAG-DTA-injected (DTA) mice at P23. Scale bar, 0.5 nA, 10 ms. b , Frequency distributions of the number of CFs innervating each PC during P23 to P47 for control (orange columns, n = 58 cells, 7 mice) and AAV-hMAG-DTA-injected (cyan, n = 66 cells, 9 mice) mice. c , Average total amplitude of CF-EPSCs (summation of all CF-EPSC in each PC) for control (orange columns) and AAV-hMAG-DTA-injected mice (cyan), with individual data points (from each cell) overlaid on the bars. d , Fluorescence microscopy images showing PCs labeled with Car8 (magenta) and climbing fiber terminals with vesicular glutamate transporter type 2 (VGluT2, green) in control and AAV-hMAG-DTA-injected mice at P21 and P72. Scale bars: 20 µm. e, Frequency distributions of the number of perisomatic CF terminals on PCs for control (orange, n = 253 cells, 3 mice at P12; n = 159 cells, 3 mice at P72-P76) and AAV-hMAG-DTA-injected mice (cyan, n = 157 cells, 3 mice at P12; n = 160 cells, 3 mice at P72-P76). f , Immunofluorescence images showing VGluT2 (green) in control and AAV-hMAG-DTA-injected mice at P21 and P72, highlighting the top dots of VGluT2 in the molecular layer (arrows) and the demarcation between dendrites and soma of each PC (dotted line). Scale bars: 20 µm. g , Bar graph summarizing the relative positioning of CF terminals to the molecular layer thickness in control versus AAV-hMAG-DTA-injected mice at P21 and P72-P82 (n = 13-18 measurements per group, from 3 mice each). Statistical significance denoted as **p < 0.01 and ****p < 0.0001, according to Mann-Whitney U tests.

Article Snippet: For AAV-hMAG-DTA, we amplified the DTA coding sequence from the pAAV-mCherry-flex-dtA (a gift from Naoshige Uchida, Addgene plasmid # 58536) using primer pairs (Table.

Techniques: Control, Injection, Fluorescence, Microscopy, Labeling, Immunofluorescence, MANN-WHITNEY

Journal: eLife

Article Title: Satellite glia modulate sympathetic neuron survival, activity, and autonomic function

doi: 10.7554/eLife.74295

Figure Lengend Snippet:

Article Snippet: Sequence-based reagent , Gt(ROSA)26Sortm1(DTA)Jpmb/J _F , The Jackson Laboratory , PCR primers , AAAGTCGCTCTGAGTTGTTAT.

Techniques: Sequencing, Control, Imaging, In Situ, Enzyme-linked Immunosorbent Assay, RNAscope, Multiplex Assay, Software

Journal: Scientific Reports

Article Title: Control of axillary bud growth in tobacco through toxin gene expression system

doi: 10.1038/s41598-021-96976-3

Figure Lengend Snippet: The alignment results of CDS sequences ( a ) and protein sequences( b ) of modified DTA and original DTA .

Article Snippet: The CDS sequence (GenBank: KY766997.1) of the toxic gene DTA was optimized and synthesized by The Beijing Genomics Institute (BGI) .

Techniques: Modification