aav5 hsyn dio mcherry  (Addgene inc)

Journal: PLOS Biology

Article Title: Thalamic reticular neurons provide cell type-specific modulation of sound processing in the auditory thalamus

doi: 10.1371/journal.pbio.3003693

Figure Lengend Snippet: A. Experimental design for anatomical experiments. PV-Cre mice were injected with AAV5-hSyn1-DIO-mCherry into TRN for anterograde tracing of PV TRN neurons. Inset: mCherry expression in the TRN indicating the localization of the virus. B. Top : projections of PV TRN neurons primarily target the ventral subdivision of the MGB. Left panel : viral expression of projections to the MGB (magenta). Middle panel : immunohistochemical labeling of calretinin in the MGB (green). Right panel : merged channels. Bottom : quantification of the fluorescence intensity of projections from PV TRN neurons that target MGB (Dashed box in top middle panel outlining the ROI). Left panel : intensity of viral expression of projections to the MGB in the ROI along the dorso-ventral axis. Dashed lines indicate extent of vMGB. Middle panel : intensity of the immunohistochemical labeling of calretinin in the MGB in the ROI along the dorso-ventral axis. Right panel : merged channels. Bar plot: mean intensity of the projections of PV TRN neurons in dorsal and ventral MGB ( N = 14 ROIs; n = 3 mice). C. SST-Cre mice were injected with AAV5-hSyn1-DIO-mCherry into TRN for anterograde tracing of SST TRN neurons. D. Top : projections of SST TRN neurons primarily avoid the ventral subdivision of the MGB. Left panel : viral expression of projections to the MGB (magenta). Middle panel : immunohistochemical labeling of calretinin in the MGB (green). Right panel : merged channels. Bottom : quantification of the fluorescence intensity of projections from SST TRN neurons that target MGB (Dashed box in top middle panel outlining the ROI). Left panel : intensity of viral expression of projections to the MGB in the ROI along the dorso-ventral axis. Middle panel : intensity of the immunohistochemical labeling of calretinin in the MGB in the ROI along the dorso-ventral axis. Right panel : merged channels. Bar plot: mean intensity of the projections of PV TRN neurons in dorsal and ventral MGB ( N = 8 ROIs; n = 3 mice). Scale bar = 0.25 mm. vMGB: ventral MGB; dMGB: dorsal MGB; mMGB: medial MGB; White dashed box in the top middle panel indicates ROI for fluorescence intensity quantification. The data underlying this Figure can be found in .

Article Snippet: For anterograde tracing of PV or SST neurons of the audTRN and control experiments we injected 400–450 nL 0f AAV5- hSyn-DIO-mCherry (≥ 7 × 1012 vg/mL, Addgene 50459-AAV5).

Techniques: Injection, Anterograde Tracing, Expressing, Virus, Immunohistochemical staining, Labeling, Fluorescence

Journal: PLOS Biology

Article Title: Thalamic reticular neurons provide cell type-specific modulation of sound processing in the auditory thalamus

doi: 10.1371/journal.pbio.3003693

Figure Lengend Snippet: A. PV-Cre mice were injected with AAV5-hSyn1-SIO-stGtACR1-FusionRed into the audTRN and were implanted with an optic fiber. We recorded from MGB neurons in awake head-fixed mice. We presented a set of pure tones in light-On and light-Off conditions. We selectively inactivated PV TRN neurons while recording from neurons in the MGB using a vertical multi-channel electrode that spanned the depth of dMGB and vMGB, with the top electrode positioned at the tip of dMGB. Left: example probe track labeled using DiD (magenta), from a recording in MGB. Center: images from Paxinos Mouse Brain Atlas with target injection and recording locations superimposed. Right: diagram of stimuli and recording setup. B. Pie chart breaking down the effect of PV TRN inactivation on neurons of the MGB. We found that 41% of recorded units had suppressed tone-evoked responses (red slice), 30% of recorded units had facilitated tone-evoked responses (blue slice), and 29% of tone-responsive units were not affected by light manipulation (gray slice). C. Distribution of significant modulations of MGB responses during inactivation of PV TRN neurons. Red: suppressed neurons; Blue: facilitated neurons. D. Scatter plot showing the change in firing rate (FR) of recorded MGB neurons with PV TRN inactivation as a function of probe depth. Dotted gray lines indicate MGB sub-region borders. E. Mean change in FR of recorded MGB neurons with PV TRN inactivation as a function of probe depth in 100 mm bins. F. Total change in FR of recorded MGB neurons with PV TRN inactivation as a function of probe depth in 100 mm bins. G–L. Same as A–F , but for SST TRN inactivation in SST-Cre mice, where we found that 58% of recorded units had suppressed tone-evoked responses (red slice), 1% of recorded units had facilitated tone-evoked responses (navy blue slice), and 41% of tone-responsive units were not affected by SST TRN inactivation (gray slice). (N (PV) = 256, n = 4 mice. N (SST): 312, n = 4 mice). ROIs; n = 3 mice). Scale bar = 0.50 mm. vMGB: ventral MGB; dMGB: dorsal MGB; mMGB: medial MGB.

Article Snippet: For anterograde tracing of PV or SST neurons of the audTRN and control experiments we injected 400–450 nL 0f AAV5- hSyn-DIO-mCherry (≥ 7 × 1012 vg/mL, Addgene 50459-AAV5).

Techniques: Injection, Labeling

Journal: PLOS Biology

Article Title: Thalamic reticular neurons provide cell type-specific modulation of sound processing in the auditory thalamus

doi: 10.1371/journal.pbio.3003693

Figure Lengend Snippet: A. PV-Cre mice were injected with AAV5-hSyn1-SIO-stGtACR1-FusionRed into the audTRN and were implanted with an optic fiber. We selectively inactivated PV TRN neurons on half of the trials while recording from MGB neurons in awake head-fixed mice in response to pure tones. On light-On trials, light was presented simultaneously with the tone. Raster plot of responses spike times for an example facilitated unit; trials are ordered by frequency and light condition. The gray box indicates light-Off conditions, and the blue box indicates the light-On conditions. B. PSTH of the facilitated example unit averaged across all trials for light-Off (black line) and light-On (blue line) conditions. C. Frequency response function for the facilitated unit during the early and late 25 ms of tone presentation during light-Off and light-On conditions. D–F. same as A–C , but for a representative suppressed neuron. G, H. Left: mean PSTH of facilitated ( G ) and suppressed ( H ) recorded neurons in MGB ( N = 74 facilitated and N = 104 suppressed neurons) for Light-Only trials (green line), tone-On, light-Off trials (black line), and tone-On, light-On trials (light blue line). Right: time to peak of tone-evoked responses for facilitated ( G , blue) and suppressed ( H , red) MGB neurons for light-Off (blue/red) and light-On (light blue) trials. I–K . Left: scatter plots and box plots of the firing rates for light-Off and light-On conditions for facilitated (blue) and suppressed (red) neurons during spontaneous activity prior to tone/light presentation (I) , 0–25 ms post tone onset (J) , and 25–50 ms post tone onset (K) . L , M. Left panels: baseline-subtracted light responses for light-Only stimuli (green), and light-induced change in firing rate between the two tone-On conditions (blue, difference between tone-On, light-On and tone-On, light-Off firing rates). Middle panel: scatter plot, and right panel: box plots of mean firing rate changes in light response in tone-On vs. tone-Off conditions for the first 0–25 ms of stimulus presentation. L : facilitated neurons. M : suppressed neurons. Dashed line is x = y. N = 256 units; n = 4 mice. Here and below, boxplots depict the median and interquartile range (IQR) for each condition; whiskers extend to values within 1.5xIQR; shaded areas in psths represent SEM±; error bars represent SEM±. *** denotes p-value < 0.001; * denotes p-value < 0.05. The data underlying this Figure can be found in .

Article Snippet: For anterograde tracing of PV or SST neurons of the audTRN and control experiments we injected 400–450 nL 0f AAV5- hSyn-DIO-mCherry (≥ 7 × 1012 vg/mL, Addgene 50459-AAV5).

Techniques: Injection, Activity Assay

Journal: PLOS Biology

Article Title: Thalamic reticular neurons provide cell type-specific modulation of sound processing in the auditory thalamus

doi: 10.1371/journal.pbio.3003693

Figure Lengend Snippet: A. SST-Cre mice were injected with AAV5-hSyn1-DIO-stGtACR1-FusionRed into the audTRN and were implanted with an optic fiber. We recorded from MGB neurons in awake head-fixed mice. We presented a set of pure tones in light-On and light-Off conditions. Light was presented simultaneous with the tone. We selectively inactivated SST TRN neurons on half of trials while recording from neurons in MGB. Raster plot of response spike times for an example facilitated unit; trials are ordered by frequency and light condition. The gray box indicates light-Off conditions, and the blue box indicates light-On conditions. B. PSTH of the facilitated example unit averaged across all trials for the light-Off (black line) and light-On (blue line) conditions. C. Frequency response function for the facilitated unit during the early and late 25 ms of tone presentation during light-Off and light-On conditions. D–F . same as A–C , but for a representative suppressed neuron. G, H. Left: mean PSTH of facilitated ( G ) and suppressed ( H ) recorded neurons in MGB ( N = 4 facilitated. N = 181 suppressed) for light-Only trials (green line), tone-On, light-Off trials (black line), and tone-On, light-On trials (light blue line). Right: time to peak of tone-evoked responses for facilitated ( G , blue) and suppressed ( H , red) MGB neurons for light-Off (blue/red) and light-On (light blue) trials. I–K. Scatter plots and box plots of mean firing rates for light-Off and light-On conditions for facilitated (blue) and suppressed (red) neurons during spontaneous activity (I), 0–25 ms post-stimulus onset (J), and 25–50 ms post-stimulus onset (K). L, M. Left panels: baseline-subtracted light responses for light-Only stimuli (green), and light-induced change in firing rate between the two tone-On conditions (blue, difference between tone-On, light-On and tone-On, light-Off firing rates). Center: scatter plot of firing rate changes in light response in tone-On vs. tone-Off conditions. Right: box plot of mean light responses in tone-Off and tone-On conditions. Dashed line is x = y. L : Facilitated neurons. M : Suppressed neurons. N = 312 units; n = 4 mice. The data underlying this Figure can be found in .

Article Snippet: For anterograde tracing of PV or SST neurons of the audTRN and control experiments we injected 400–450 nL 0f AAV5- hSyn-DIO-mCherry (≥ 7 × 1012 vg/mL, Addgene 50459-AAV5).

Techniques: Injection, Activity Assay