aav1  (Addgene inc)

Journal: eLife

Article Title: Serotonin modulates infraslow oscillation in the dentate gyrus during non-REM sleep

doi: 10.7554/eLife.100196

Figure Lengend Snippet: ( A ) Representative recording session showing infraslow oscillation (ISO) during NREM sleep. From top to bottom: brain states, EEG power spectrogram (0–25 Hz), EMG amplitude, photometric signal. ( B ) Left, Schematic representation of the recording setup. Right, a fluorescence image showing the expression of GCaMP6s (green) in the granule cell layer and the optic fiber placement (dashed line) in a Dock10 Cre mouse. Scale bar, 500 µm. ( C ) Quantification of calcium activity in granule cells (13 recording sessions in 5 Dock10 Cre mice, n.s. – no significance, **p<0.01, paired t-test) in wake (W), NREM sleep (N), and rapid eye movement (REM) sleep (R). ( D ) Left: oscillation peak frequency during NREM sleep and wake based on Fourier transformation of the photometry signal. Right: Quantification of calcium oscillations in granule cells (GCs) (13 sessions in 5 mice). ( E ) A representative example showing the coincidence of calcium troughs (indicated by *) with microarousals (MAs). ( F ) Percentage of state transition outcome from each calcium dip. ( G ) Peri-stimulus time histogram (PSTH) in one recording session showing calcium signal aligned with the onset of MAs. Bottom left: parameters of the calcium signal used for quantification. ( H ) Quantification of the latency (t) and magnitude of the calcium trough (Drop) during MAs (13 sessions from 5 Dock10 Cre mice).

Article Snippet: Strain (pAAV.Syn.Flex.GCaMP6s.WPRE.SV40) , AAV1-SYN-FLEX-GCaMP6s , Addgene , RRID: Addgene_100845 , .

Techniques: Fluorescence, Expressing, Activity Assay, Transformation Assay

Journal: eLife

Article Title: Serotonin modulates infraslow oscillation in the dentate gyrus during non-REM sleep

doi: 10.7554/eLife.100196

Figure Lengend Snippet: ( A ) Representative recording session showing infraslow calcium oscillation during NREM sleep in a Drd2 Cre+/- mouse injected with AAV-FLEX-GCaMP6s. From top to bottom: brain states, EEG power spectrogram (0–25 Hz), EMG amplitude, photometric signal. ( B ) Fluorescence images showing the expression of GCaMP6s (green) in the mossy cells in the area marked with the white rectangle. Blue, DAPI. Scale bars, 500 µm and 50 µm. ( C ) Quantification of calcium activity in mossy cells during different brain states (12 recording sessions in 4 Drd2 Cre+/- mice; n.s. – no significance, **p<0.01, paired t-test). ( D ) Left: oscillation peak frequency during NREM sleep based on Fourier transformation of the photometry signal. Right: quantification of calcium oscillations during NREM sleep. ( E ) A representative example showing the coincidence of calcium troughs (indicated by *) with microarousals (MAs) (represented with vertical green lines). ( F ) Percentage of state transition outcome from each calcium dip. ( G ) Peri-stimulus time histogram (PSTH) from one recording session showing calcium signals (red) and EMG signals (light green) aligned with the onset of MAs. ( H ) Quantification of the latency (t) and the magnitude of calcium troughs (Drop) during the MAs (12 sessions from 4 Drd2 Cre mice).

Article Snippet: Strain (pAAV.Syn.Flex.GCaMP6s.WPRE.SV40) , AAV1-SYN-FLEX-GCaMP6s , Addgene , RRID: Addgene_100845 , .

Techniques: Injection, Fluorescence, Expressing, Activity Assay, Transformation Assay

Journal: eLife

Article Title: Serotonin modulates infraslow oscillation in the dentate gyrus during non-REM sleep

doi: 10.7554/eLife.100196

Figure Lengend Snippet: ( A ) Left, Schematic representation of the two-site photometry experimental design. Right, Expression of CaMKII-GCaMP6s and fiber placement in the dentate gyrus (DG) and raphe nuclei. Scale bars, 500 µm. ( B ) A representative example of concurrent recording of DG and raphe 5-HT neurons in a Slc6a4 Cre+/- mouse during sleep. From top to bottom: brain states, EEG power spectrogram (0–25 Hz), EMG amplitude, photometric calcium signals (CaMKII-G6s) in DG and in dorsal raphe. ( C ) Correlation analysis of calcium activity between DG and raphe 5-HT neurons during NREM sleep and wakefulness in one recording session. ( D ) Quantification of correlation coefficient between DG activity and raphe activity during different brain states (11 sessions from 3 Slc6a4 Cre+/- mice, ***p<0.001, paired t-test).

Article Snippet: Strain (pAAV.Syn.Flex.GCaMP6s.WPRE.SV40) , AAV1-SYN-FLEX-GCaMP6s , Addgene , RRID: Addgene_100845 , .

Techniques: Expressing, Activity Assay

Journal: eLife

Article Title: Serotonin modulates infraslow oscillation in the dentate gyrus during non-REM sleep

doi: 10.7554/eLife.100196

Figure Lengend Snippet: ( A ) Representative recording session showing calcium activity in a wild-type mouse injected with AAV9-CaMKII-GCaMP6s in the DG. From top to bottom: brain states, EEG power spectrogram (0–25 Hz), EMG, photometric signal. ( B ) A fluorescent image showing the constrained expression of GCaMP6s (green) in the granule cell layer of the DG. Scale bar, 500 µm. ( C ) Quantification of calcium activity (16 recording sessions in 6 C57BL/6 J mice, n.s., no significance, **p<0.01, paired t-test) in wake (W), NREM sleep (N), and REM sleep (R). ( D ) Left: oscillation peak frequency during NREM sleep and wake based on Fourier transformation of the photometry signal. Right: Quantification of calcium oscillations in CaMKII-labeled cells (16 sessions in 6 mice).

Article Snippet: Strain (pAAV.Syn.Flex.GCaMP6s.WPRE.SV40) , AAV1-SYN-FLEX-GCaMP6s , Addgene , RRID: Addgene_100845 , .

Techniques: Activity Assay, Injection, Expressing, Transformation Assay, Labeling

Journal: eLife

Article Title: Serotonin modulates infraslow oscillation in the dentate gyrus during non-REM sleep

doi: 10.7554/eLife.100196

Figure Lengend Snippet: ( A ) Schematic representation of the experimental design. A mix of AAV9-CamKII-GCaMP6s and AAV1-hSyn-Cre was injected into the DG of Htr1a flox/flox mice. ( B ) Representative example showing photometry and EEG recordings in the DG of a control mouse injected with AAV9-CaMKII-GCaMP6s alone. Right, Fourier transformation of calcium activity during wake (blue) non-rapid eye movement (NREM) sleep (red). ( C ) A representative example showing photometry and EEG recordings in the DG of a mouse injected with AAV9-CaMKII-GCaMP6s and AAV1-hSyn-Cre. Right, Fourier transformation of calcium activity during wake (blue) NREM sleep (red). ( D ) Left, Quantification of the relative power of the calcium oscillation in the range of 1–2 cycles/min in the Cre and control groups (16 sessions from 5 mice for Cre, 16 sessions from 6 mice for control). Right, Quantification of calcium oscillation amplitudes in the Cre and control groups. Calcium signals in each mouse were normalized to Z scores. **p<0.01, ***p<0.001, unpaired t-test. ( E ) Schematic representation of the contextual fear conditioning (CFC) experimental design. ( F ) Left, Contextual fear recall tests showing percentage of freezing in 1 min time bins for Htr1a flox+/+ mice bilaterally injected with AAV9-CaMKII-Cre-GFP (Cre) or AAV9-CaMKII-GFP (GFP). Right, Quantification of freezing behavior over 5 min interval during contextual recall tests in Cre and GFP groups (N=11 for Cre, N=12 for GFP, *p<0.05, ***p<0.001, unpaired t-test).

Article Snippet: Strain (pAAV.Syn.Flex.GCaMP6s.WPRE.SV40) , AAV1-SYN-FLEX-GCaMP6s , Addgene , RRID: Addgene_100845 , .

Techniques: Injection, Control, Transformation Assay, Activity Assay

Journal: eLife

Article Title: Serotonin modulates infraslow oscillation in the dentate gyrus during non-REM sleep

doi: 10.7554/eLife.100196

Figure Lengend Snippet:

Article Snippet: Strain (pAAV.Syn.Flex.GCaMP6s.WPRE.SV40) , AAV1-SYN-FLEX-GCaMP6s , Addgene , RRID: Addgene_100845 , .

Techniques: Generated, Plasmid Preparation, Virus, RNAscope

Journal: Cell reports

Article Title: Higher-order thalamic input to cortex selectively conveys state information

doi: 10.1016/j.celrep.2025.115292

Figure Lengend Snippet: (A) Schematic of pathways originating from the higher-order visual thalamus (lateral posterior nucleus [LP]) and multiple visual cortical regions that project to the higher-order posterior medial (PM) visual cortex. (B) Coronal sections containing thalamocortical and corticocortical projection neurons retrogradely labeled via injection of CAV2-Cre into the PM of tdTomato reporter (Ai9) mice. Cells labeled by tdTomato send projections to the PM (dLGN, dorsal lateral geniculate nucleus; RL, rostral lateral visual cortex; AL: anterior lateral visual cortex; LM, lateral medial visual cortex). Scale bar: 500 μm. (C) Quantification of overall cell density for thalamocortical and corticocortical projection neurons sending axons to the PM. Dotted lines connect values from individual Ai9 animals ( N = 10 mice). (D) Quantification of cell counts (normalized to the histological section with the highest cell count in each animal) along the anterior-posterior axis of the brain (relative to bregma) for each thalamocortical and corticocortical projection neuron type. Numbered arrows correspond to (B 1 )–(B 4 ) ( N = 10 mice). (E) Thalamocortical (LP→PM) and corticocortical (V1→PM, LM→PM) projection axons in the PM anterogradely labeled with GCaMP6s via adeno-associated virus (AAV) injection in the corresponding presynaptic regions. Scale bar: 50 μm. (F) Laminar distribution of axonal fluorescence intensity for each projection type ( N = 4 mice per projection type). (G) Top: schematic of in vivo imaging setup. Bottom: image from video monitoring of the mouse’s facial motion and pupil size. (H) Fluorescence traces of Ca 2+ activity from individual cell body (PM) and axon (V1→PM, LM→PM, and LP→PM) regions of interest (ROIs) imaged simultaneously with behavioral-state monitoring (locomotion speed, facial motion, and pupil size) and visual stimuli presentation. Error bars denote SEM.

Article Snippet: To express the calcium indicator GCaMP6s in neuronal cell bodies or long-range projection axons either AAV5-Syn-GCaMP6s or AAV1-Syn-GCaMP6s (1×10 13 gc/mL; Addgene #100843) was injected into the relevant brain region.

Techniques: Labeling, Injection, Cell Counting, Virus, Fluorescence, In Vivo Imaging, Activity Assay

Journal: Cell reports

Article Title: Higher-order thalamic input to cortex selectively conveys state information

doi: 10.1016/j.celrep.2025.115292

Figure Lengend Snippet: (A) Schematic of method for simultaneously monitoring the neuronal activity of PM cell bodies and long-range projection axons terminating in the PM based on expression of GCaMP6s in long-range axon terminals and ribo-GCaMP6m in PM cell bodies. (B) In vivo field of view with PM cell bodies expressing riboGCaMP6m and LM axon terminals expressing GCaMP6s. (C) Fluorescence traces of Ca 2+ activity simultaneously recorded in PM cell bodies expressing riboGCaMP6m and projection axons expressing GCaMP6s. (D) Mean Ca 2+ event rates among the different cell types. (V1→PM: N = 5 animals and n = 332 ROIs; LM→PM: N = 4 animals and n = 211 ROIs; LP→PM: N = 5 animals and n = 191 ROIs; and PM: N = 14 animals and n = 168 ROIs). (E) Ca 2+ event rates for each afferent population aligned to simultaneously recorded Ca 2+ events in PM neurons (raw data minus data from shuffled PM event times). (F) Peak Ca 2+ event rates aligned to PM neuron Ca 2+ events (V1→PM: N = 5 animals and n = 59 PM ROIs; LM→PM: N = 4 animals and n = 53 PM ROIs; LP→PM: N = 5 animals and n = 56 PM ROIs; and PM activity aligned with PM events: N = 14 animals and n = 168 PM ROIs). * p < 0.05, ** p < 0.01, and *** p < 0.001; semi-weighted t test; Benjamini-Hochberg correction for false discovery rate. Error bars denote SEM.

Article Snippet: To express the calcium indicator GCaMP6s in neuronal cell bodies or long-range projection axons either AAV5-Syn-GCaMP6s or AAV1-Syn-GCaMP6s (1×10 13 gc/mL; Addgene #100843) was injected into the relevant brain region.

Techniques: Activity Assay, Expressing, In Vivo, Fluorescence

Journal: Cell reports

Article Title: Higher-order thalamic input to cortex selectively conveys state information

doi: 10.1016/j.celrep.2025.115292

Figure Lengend Snippet: (A) Schematic of viral injections for co-expressing GCaMP6s in PM cortical neurons and the Cre-dependent inhibitory opsin eOPN3 in presynaptic projection axon terminals. (B) Left: expression of eOPN3-mScarlet (red) in thalamocortical neurons in the LP. Right: expression of GCaMP6s in PM cortical neurons (green) and eOPN3-mScarlet (red) in the surrounding LP thalamocortical terminals. Scale bar: 30 μm. (C) Visual contrast-response curves of PM neurons in control animals and animals with eOPN3 expressed in corticocortical and higher-order thalamocortical axons for sessions with (light-emitting diode [LED]) and without (dark) optogenetic stimulation. (D) Scatterplots showing visual response magnitudes from individual PM neurons identified during both dark and LED imaging sessions. (E) As in (C) but for cross-correlations between PM neuronal activity and pupil size. (F) As in (D) but for peak correlation values between neuronal activity and pupil size. (G) Differences between visual response magnitudes during sessions with and without optogenetic stimulation (control: N = 5 animals and n = 195 ROIs; V1→PM eOPN3: N = 7 animals and n = 227 ROIs; LM→PM eOPN3: N = 4 animals and n = 215 ROIs; and LP→PM eOPN3: N = 6 animals and n = 327 ROIs) (control min./max. val. = −3.2/2.5; V1→PM eOPN3 min./max. val. = −5.3/3.1; LM→PM eOPN3 min./max. val. = −3.1/5.0; and LP→PM eOPN3 min./max. val. = −3.1/1.0). (H) As in (G) but for differences in peak ΔF/F-pupil correlation values between sessions with and without optogenetic stimulation (control: N = 7 animals and n = 327 ROIs; V1→PM eOPN3: N = 4 animals and n = 169 ROIs; LM→PM eOPN3: N = 3 animals and n = 167 ROIs; and LP→PM eOPN3: N = 5 animals and n = 468 ROIs). * p < 0.05, ** p < 0.01, and *** p < 0.001; semi-weighted t test; Benjamini-Hochberg correction for false discovery rate. Error bars denote SEM.

Article Snippet: To express the calcium indicator GCaMP6s in neuronal cell bodies or long-range projection axons either AAV5-Syn-GCaMP6s or AAV1-Syn-GCaMP6s (1×10 13 gc/mL; Addgene #100843) was injected into the relevant brain region.

Techniques: Expressing, Control, Imaging, Activity Assay

Journal: Cell reports

Article Title: Higher-order thalamic input to cortex selectively conveys state information

doi: 10.1016/j.celrep.2025.115292

Figure Lengend Snippet:

Article Snippet: To express the calcium indicator GCaMP6s in neuronal cell bodies or long-range projection axons either AAV5-Syn-GCaMP6s or AAV1-Syn-GCaMP6s (1×10 13 gc/mL; Addgene #100843) was injected into the relevant brain region.

Techniques: Virus, Plasmid Preparation, Software