virus expressing gcamp6s36  (Addgene inc)

Journal: bioRxiv

Article Title: Neural responses to social cues in the accessory olfactory bulb are altered by context and experience

doi: 10.1101/2023.11.28.569126

Figure Lengend Snippet: Transgenic and viral strategy to record from AOB projection neurons during social interactions. (A) The AOB receives input directly from the VNO and makes, largely reciprocal, connections with subcortical brain regions devoted to social behavior (red arrows; BNST: bed nucleus of the stria terminalis; ACo: cortical amygdaloid nucleus; MeA: medial amygdala; PMCo: posteromedial cortical amygdala). (Inset) Mitral and tufted neurons are the projection neurons from AOB to the social behavior network (SBN; Glom: glomerular layer; PG: periglomerular cells; M/T: mitral and tufted cells; GC: granule cells). (B) Fiberphometry recordings from AOB mitral and tufted cells rely on conditional AAV-driven expression of GCaMP in a Tbet:Cre mouse paired with a fiberoptic probe implanted in the AOB . (C) Histology showing accurate GCaMP expression (cyan) in M/T cells.

Article Snippet: We injected a conditional GCaMP expressing AAV virus (AAV9:FLEX:GCaMP6s; Addgene Plasmid #:100845; ) in the AOB (Bregma 3.5, Lateral 1.0, Depth 1.0) of Tbet-cre mice to target M/T cells in the AOB.

Techniques: Transgenic Assay, Expressing

Journal: bioRxiv

Article Title: Neural responses to social cues in the accessory olfactory bulb are altered by context and experience

doi: 10.1101/2023.11.28.569126

Figure Lengend Snippet: Strategy for automating social interactions during fiberphotometric recording. (A) Trace of mouse position of over 30 seconds (color: intensity of AOB GCaMP signal; M: male; F: female; P: predator; C: control). (B) Social stimuli were presented once every 2 minutes for a 1-minute duration and behavior states were manually scored (exploratory behavior, social interaction, approach, and investigation). (C) Top: density plots indicating the location of an animal during the time a stimulus is available (Left; all time; center left: male stimulus; center: female; center right: predator; right; empty cup). Bottom: the average time (percent of total) spent investigating a stimulus when the stimulus is available (open) or not available (closed; asterisks indicate p<0.0001, paired t-test). (D) Percentage of stimulus presentations that resulted in a successful investigation by the experimental animal (error bars: SEM). (E) Fraction of stimulus presentation events that resulted in a successful investigation event for 7 successive presentations of each stimulus (3 different randomly interleaved stimuli; 21 total) when there was not a predator (left) or was a predator (right) in the stimulus set (red: female; blue: male; green: predator; grey: empty cup; shaded regions: SEM).

Article Snippet: We injected a conditional GCaMP expressing AAV virus (AAV9:FLEX:GCaMP6s; Addgene Plasmid #:100845; ) in the AOB (Bregma 3.5, Lateral 1.0, Depth 1.0) of Tbet-cre mice to target M/T cells in the AOB.

Techniques: Control

Journal: bioRxiv

Article Title: Neural responses to social cues in the accessory olfactory bulb are altered by context and experience

doi: 10.1101/2023.11.28.569126

Figure Lengend Snippet: Example of sensory responses of AOB neurons to social and predator stimuli in an adult female mouse. (A) Video frames surrounding a social interaction, with 0 seconds (center) aligned to the onset of stimulus investigation (Blue shading indicates the availability of the male stimulus). The predator (P) and female (F) stimuli were closed during the duration of the shown frames (bracket indicates the investigation event in panel B highlighted in panel A). (B) GCaMP signal (black line; z-score normalized) during seven randomized presentations of three stimuli (male, female, predator; order indicated by the colored boxes). Vertical dashed lines indicate the start of all investigations of the male stimulus. (C) Average responses (from panel B) aligned to the start of investigation of male, female, and predator stimuli (shading: SEM).

Article Snippet: We injected a conditional GCaMP expressing AAV virus (AAV9:FLEX:GCaMP6s; Addgene Plasmid #:100845; ) in the AOB (Bregma 3.5, Lateral 1.0, Depth 1.0) of Tbet-cre mice to target M/T cells in the AOB.

Techniques:

Journal: bioRxiv

Article Title: Neural responses to social cues in the accessory olfactory bulb are altered by context and experience

doi: 10.1101/2023.11.28.569126

Figure Lengend Snippet: Average GCaMP response across stimulus presentations. (A) Average GCaMP signal of female (red), male (blue), predator (green) and control (grey) stimuli relative to the first investigation after a stimulus became available (shading: SEM; N = 16: 7 males, 9 females). (B). Average z-score during the 20 seconds following sensory investigation for female, male, predator, and control stimuli. Error bars: SEM; asterisks: p-value<0.000001.

Article Snippet: We injected a conditional GCaMP expressing AAV virus (AAV9:FLEX:GCaMP6s; Addgene Plasmid #:100845; ) in the AOB (Bregma 3.5, Lateral 1.0, Depth 1.0) of Tbet-cre mice to target M/T cells in the AOB.

Techniques: Control

Journal: bioRxiv

Article Title: Neural responses to social cues in the accessory olfactory bulb are altered by context and experience

doi: 10.1101/2023.11.28.569126

Figure Lengend Snippet: AOB response to social stimuli depends on context. (A) Left, average GCaMP signal (z-score normalized) across presentations of female stimuli during experiments that included predator stimuli (red) or did not include predator stimuli (grey). Right, mean GCaMP response to female stimuli during predator and no predator experiments (t-test comparison p value = 0.011, error bars: SEM). (B) Pseudocolored plot of the average response to female stimuli for each of 7 presentations during the predator condition (top) or no predator condition (bottom). (C) Effect of repeated presentations of female stimuli on the average GCaMP response (0-20 seconds) for predator (red) and no predator (gray; error bars: SEM). (D) Left, average GCaMP signal (z-score normalized) across presentations of male stimuli during experiments that included predator stimuli (blue) or did not include predator stimuli (grey). Right, mean GCaMP response to male stimuli during predator and no predator experiments (t-test comparison p value = 0.018; error bars: SEM). (E) Pseudocolored plot of the average response to male stimuli for each of 7 presentations during the predator condition (top) or no predator condition (bottom). (F) Effect of repeated presentations of male stimuli on the average GCaMP response (0-20 seconds) for predator (blue) and no predator (gray; error bars: SEM).

Article Snippet: We injected a conditional GCaMP expressing AAV virus (AAV9:FLEX:GCaMP6s; Addgene Plasmid #:100845; ) in the AOB (Bregma 3.5, Lateral 1.0, Depth 1.0) of Tbet-cre mice to target M/T cells in the AOB.

Techniques: Comparison

Journal: bioRxiv

Article Title: Neural responses to social cues in the accessory olfactory bulb are altered by context and experience

doi: 10.1101/2023.11.28.569126

Figure Lengend Snippet: AOB responses convey stimulus novelty on multiple time scales. (A) GCaMP signal (black line; z-score normalized) during an initial baseline period followed by 18 presentations of stimulus A and 3 presentations of stimulus B presented in a standardized order shown by the boxes below. Vertical dashed red lines indicate investigation events to the less frequently presented stimulus. (B) Pseudocolor plot representing the average GCaMP signal aligned to the start of a sensory investigation. The stimulus order is as shown in panel A and changes between the presented stimulus are represented by white space. Purple asterisks represent novel trials. (C) Left, average GCaMP signal across presentations to rare (stimulus B: red) or frequent (stimulus A: gray) age and sex matched social stimuli. Right, mean GCaMP response to rare stimuli and frequent stimuli. (D) Average AOB GCaMP response to frequent (grey) and rare (red) stimuli during the 20 seconds following stimulus investigation (error bars: SEM). (E) Left, average GCaMP signal across presentations to novel (first presentation of either stimulus A or B: purple; asterisks in A,B, D) or familiar (subsequent presentations of either stimulus A or B: gray) age and sex matched social stimuli. Right, mean GCaMP response to novel (N) stimuli and familiar (F) stimuli.

Article Snippet: We injected a conditional GCaMP expressing AAV virus (AAV9:FLEX:GCaMP6s; Addgene Plasmid #:100845; ) in the AOB (Bregma 3.5, Lateral 1.0, Depth 1.0) of Tbet-cre mice to target M/T cells in the AOB.

Techniques:

Journal: bioRxiv

Article Title: Neural responses to social cues in the accessory olfactory bulb are altered by context and experience

doi: 10.1101/2023.11.28.569126

Figure Lengend Snippet: Analysis pipeline for fiber photometry signals and behavior. (A) GCaMP and isosbestic signal are collected and synced with behavior (left). GCaMP and isosbestic signals are separated utilizing lock-in sampling and amplification (middle). GCaMP and isosbestic signals are compared via regression normalization allowing the isolation of calcium dependent AOB signals from movement. (B) Scored behavioral states and events are synced with calcium-dependent AOB signal z-scores.

Article Snippet: We injected a conditional GCaMP expressing AAV virus (AAV9:FLEX:GCaMP6s; Addgene Plasmid #:100845; ) in the AOB (Bregma 3.5, Lateral 1.0, Depth 1.0) of Tbet-cre mice to target M/T cells in the AOB.

Techniques: Sampling, Amplification, Isolation