sh shank3 Search Results


sh shank3  (Vector Biolabs)
95
Vector Biolabs sh shank3
(a, d) Schema of injection sites in the NAc with AAV-scr <t>Shank3</t> -GFP or AAV-sh Shank3 -luczsGreen in ≤P6 mice (e) or at P90 (h). (a’, d’) Representative images of injection sites (scale bar: 500 µm). (b, c, e, f) Left: time spent around the enclosures during the social preference test for mice injected at ≤P6 or at P90 ((b); t (12) = 6.092, p < 0.001, (c); t (9) = 0.409, p = 0.697, (e); t (9) = 3.806, p = 0.004, (f); t (6) = 6.970, p < 0.001). Right: juvenile preference index for mice injected at ≤P6 or at P90 (one-sample t-tests against chance level = 0.5: (b), scr Shank3 ; t (12) = 5.847, p < 0.001, (c), sh Shank3 ; t (9) = 0.273, p = 0.791; (e), scr Shank3 ; t (9) = 3.928, p = 0.003, (f), shShank3; t (6) = 7.996, p < 0.001). Error bars report SEM.
Sh Shank3, supplied by Vector Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 95 stars, based on 1 article reviews
sh shank3 - by Bioz Stars, 2026-03
95/100 stars
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shank3 polyclonal antibody, abby fluor 647 conjugated  (Bioss)
N/A
Major scaffold postsynaptic density protein which interacts with multiple proteins and complexes to orchestrate the dendritic spine and synapse formation, maturation and maintenance. Interconnects receptors of the postsynaptic membrane including NMDA-type and metabotropic glutamate receptors
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shank3 antibody (s69)  (Novus Biologicals)
N/A
The SHANK3 Antibody (S69) from Novus is a SHANK3 antibody to SHANK3. This antibody reacts with Human, Mouse, Rat. The SHANK3 antibody has been validated for the following applications: Western Blot, Immunohistochemistry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation,
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shank3 antibody [alexa fluor® 405]  (Novus Biologicals)
N/A
The SHANK3 Antibody [Alexa Fluor® 405] from Novus is a SHANK3 antibody to SHANK3. This antibody reacts with Human, Mouse. The SHANK3 antibody has been validated for the following applications: Western Blot, Immunoprecipitation.
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shank3 antibody [alexa fluor® 647]  (Novus Biologicals)
N/A
The SHANK3 Antibody [Alexa Fluor® 647] from Novus is a SHANK3 antibody to SHANK3. This antibody reacts with Human, Mouse. The SHANK3 antibody has been validated for the following applications: Western Blot, Immunoprecipitation.
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shank3 antibody [dylight 488]  (Novus Biologicals)
N/A
The SHANK3 Antibody [DyLight 488] from Novus is a SHANK3 antibody to SHANK3. This antibody reacts with Human, Mouse. The SHANK3 antibody has been validated for the following applications: Western Blot, Immunoprecipitation.
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shank1 shank3 antibody atto 565  (Rockland Immunochemicals)
N/A
Anti SHANK1 SHANK3 MOUSE Monoclonal Antibody ATTO 565 Conjugated 200 358 W72
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shank3 antibody  (Rockland Immunochemicals)
N/A
Anti SHANK3 MOUSE Monoclonal Antibody 200 301 G45
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shank3 nm 033517 human tagged orf clone  (OriGene)
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SHANK3 GFP tagged Homo sapiens SH3 and multiple ankyrin repeat domains 3 SHANK3
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shank 3 antibody  (Santa Cruz Biotechnology)
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Shank 3 Antibody C 4 is a mouse monoclonal IgG1 kappa light chain provided at 200 µg ml raised against amino acids 1431 1590 mapping near the C terminal of isoform 2 of Shank 3
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anti shank3 center region polyclonal antibody  (Creative BioMart)
N/A
At least three alternatively spliced transcript isoforms of SHANK3 are known to exist Seems to be an adapter protein in the postsynaptic density PSD of excitatory synapses that interconnects receptors of the postsynaptic membrane including
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shank3 mouse monoclonal antibody  (OriGene)
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SHANK3 mouse monoclonal antibody clone N367 51
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Image Search Results


(a, d) Schema of injection sites in the NAc with AAV-scr Shank3 -GFP or AAV-sh Shank3 -luczsGreen in ≤P6 mice (e) or at P90 (h). (a’, d’) Representative images of injection sites (scale bar: 500 µm). (b, c, e, f) Left: time spent around the enclosures during the social preference test for mice injected at ≤P6 or at P90 ((b); t (12) = 6.092, p < 0.001, (c); t (9) = 0.409, p = 0.697, (e); t (9) = 3.806, p = 0.004, (f); t (6) = 6.970, p < 0.001). Right: juvenile preference index for mice injected at ≤P6 or at P90 (one-sample t-tests against chance level = 0.5: (b), scr Shank3 ; t (12) = 5.847, p < 0.001, (c), sh Shank3 ; t (9) = 0.273, p = 0.791; (e), scr Shank3 ; t (9) = 3.928, p = 0.003, (f), shShank3; t (6) = 7.996, p < 0.001). Error bars report SEM.

Journal: bioRxiv

Article Title: Inhibition of TRPV4 rescues circuit and social deficits unmasked by acute inflammatory response in a Shank3 mouse model of Autism

doi: 10.1101/2021.10.13.464215

Figure Lengend Snippet: (a, d) Schema of injection sites in the NAc with AAV-scr Shank3 -GFP or AAV-sh Shank3 -luczsGreen in ≤P6 mice (e) or at P90 (h). (a’, d’) Representative images of injection sites (scale bar: 500 µm). (b, c, e, f) Left: time spent around the enclosures during the social preference test for mice injected at ≤P6 or at P90 ((b); t (12) = 6.092, p < 0.001, (c); t (9) = 0.409, p = 0.697, (e); t (9) = 3.806, p = 0.004, (f); t (6) = 6.970, p < 0.001). Right: juvenile preference index for mice injected at ≤P6 or at P90 (one-sample t-tests against chance level = 0.5: (b), scr Shank3 ; t (12) = 5.847, p < 0.001, (c), sh Shank3 ; t (9) = 0.273, p = 0.791; (e), scr Shank3 ; t (9) = 3.928, p = 0.003, (f), shShank3; t (6) = 7.996, p < 0.001). Error bars report SEM.

Article Snippet: Viruses used in this study: (1) purified scr Shank3 and sh Shank3 (AAV1-GFP-U6-scrmbshRNA; titer: 5.9×10 13 GC/mL and AAV5-ZacF-U6-luczsGreen-sh Shank3 ; titer: 7.4×10 13 GC/mL, VectorBioLab); (2) AAV5/hsyn-DIO-hM4D(Gi)-mCherry (AV44961, titer: 5.5×10 12 virus molecules/mL, UNC GTC vector core).

Techniques: Injection

(a) Schema of injection sites in the NAc with AAV-scrShank3-GFP or AAV-shShank3-luczsGreen in ≤P6 or P90 mice. Subsequently, the NAc was dissected and mRNA was extracted. (b) Real-time PCR analysis of NAc dissected from P6- or P90-injected mice confirm the downregulation of Shank3 in sh infected mice (two-way ANOVA followed by Bonferroni’s multiple comparisons test: Virus main effect F (1, 8) = 18.80, p=0.003 ). (c, h) Time spent in compartments for mice injected ≤ P6 ((c) Paired-samples t-tests for object- vs. social-containing chambers: scrShank3; t (12) = - 5.047, p < 0.001), shShank3; t (9) = - 0.645, p =0.535, (h) Paired-samples t-tests for object- vs. social-containing chambers: scrShank3; t (9) = 3.144, p = 0.012, shShank3; t (6) = 5.686, p = 0.001). (d) Total exploration time around the enclosures for mice injected neonatally (Mann-Whitney test, p = 0.784). (e) Time spent around the enclosure containing the social stimulus ( t (21) = 2.152, p = 0.043). (f) Time spent around the non-social target ( t (21) = -3.499, p = 0.002). (g) Distance moved in the apparatus ( t (21) = -0.483, p = 0.634). (i) Total exploration time around the enclosures for mice injected during adulthood ( t (15) = 1.347, p = 0.198). (j) Time spent around the enclosure containing the social stimulus ( t (15) = -1.541, p = 0.144). (k) Time spent around the non-social target ( t (15) = 1.347, p = 0.198). (l) Distance moved in the apparatus ( t (15) = -1.544, p = 0.143). Error bars report SEM.

Journal: bioRxiv

Article Title: Inhibition of TRPV4 rescues circuit and social deficits unmasked by acute inflammatory response in a Shank3 mouse model of Autism

doi: 10.1101/2021.10.13.464215

Figure Lengend Snippet: (a) Schema of injection sites in the NAc with AAV-scrShank3-GFP or AAV-shShank3-luczsGreen in ≤P6 or P90 mice. Subsequently, the NAc was dissected and mRNA was extracted. (b) Real-time PCR analysis of NAc dissected from P6- or P90-injected mice confirm the downregulation of Shank3 in sh infected mice (two-way ANOVA followed by Bonferroni’s multiple comparisons test: Virus main effect F (1, 8) = 18.80, p=0.003 ). (c, h) Time spent in compartments for mice injected ≤ P6 ((c) Paired-samples t-tests for object- vs. social-containing chambers: scrShank3; t (12) = - 5.047, p < 0.001), shShank3; t (9) = - 0.645, p =0.535, (h) Paired-samples t-tests for object- vs. social-containing chambers: scrShank3; t (9) = 3.144, p = 0.012, shShank3; t (6) = 5.686, p = 0.001). (d) Total exploration time around the enclosures for mice injected neonatally (Mann-Whitney test, p = 0.784). (e) Time spent around the enclosure containing the social stimulus ( t (21) = 2.152, p = 0.043). (f) Time spent around the non-social target ( t (21) = -3.499, p = 0.002). (g) Distance moved in the apparatus ( t (21) = -0.483, p = 0.634). (i) Total exploration time around the enclosures for mice injected during adulthood ( t (15) = 1.347, p = 0.198). (j) Time spent around the enclosure containing the social stimulus ( t (15) = -1.541, p = 0.144). (k) Time spent around the non-social target ( t (15) = 1.347, p = 0.198). (l) Distance moved in the apparatus ( t (15) = -1.544, p = 0.143). Error bars report SEM.

Article Snippet: Viruses used in this study: (1) purified scr Shank3 and sh Shank3 (AAV1-GFP-U6-scrmbshRNA; titer: 5.9×10 13 GC/mL and AAV5-ZacF-U6-luczsGreen-sh Shank3 ; titer: 7.4×10 13 GC/mL, VectorBioLab); (2) AAV5/hsyn-DIO-hM4D(Gi)-mCherry (AV44961, titer: 5.5×10 12 virus molecules/mL, UNC GTC vector core).

Techniques: Injection, Real-time Polymerase Chain Reaction, Infection, Virus, MANN-WHITNEY

(a) Experimental design. Drd1a-dTomato mice were injected neonatally in the NAc with scr or sh Shank3 virus and whole-cell patch clamp recordings were performed during early adulthood. (a’) Representative images of the NAc of a D1R-tom+::sh Shank3 mouse (scale bar: 50 µm). (b) Example traces at 300 pA depolarizing current injection in D1R-tom+ MSNs infected with scr Shank3 (left) or with sh Shank3 (right). (c) Number of action potentials (nAPs) across increasing depolarizing current steps (0-500 pA) for D1R-tom+::scr Shank3 and sh Shank3 MSNs, in presence of picrotoxin and kynurenic acid. (Repeated measures ANOVA, virus main effect F (1,14) = 10.88, p = 0.005, current steps main effect F (10, 140) = 7.727, p < 0.001, scr Shank3 n = 8 cells, 3 mice, sh Shank3 n = 8 cells, 3 mice). (d) Example traces at 300 pA depolarizing current injection in D1R-tom-MSNs infected with scr Shank3 (left) or with sh Shank3 (right). (e) Number of action potential (nAPs) across increasing depolarizing current steps (0-500 pA) for D1R-tom-::scr Shank3 and sh Shank3 MSNs, in presence of picrotoxin and kynurenic acid. (Repeated measure (RM) two-way ANOVA, virus main effect F (1,18) = 0.098, p = 0.758, current steps main effect F (10 , 180) = 14.58, p < 0.001, n = 8 cells, 3 mice (sh), n=12 cells, 3 mice (scr)). (f) Experimental design. D1R-Cre positive (D1R:Cre + ) and negative (D1R:Cre - ) mice were injected neonatally in the NAc with scr or sh Shank3 virus and after P30 with AAV-hSyn-DIO-hM4Di-mCherry (DREADD). After 4 weeks, allowing for virus expression, the mice underwent social behaviour assessment in the three-chamber task. Mice were intraperitoneally injected with CNO 30 min before starting the test. (f’) Representative images of the NAc of a D1R:Cre + mouse infected with sh Shank3 and DREADD viruses (scale bar: 50 µm). (g, h, i, j ) Left: time around the target during the social preference test for D1R:Cre - ::scr Shank3 mice (g; t (7) = 5.453, p = 0.001); D1R:Cre + ::scr Shank3 mice (h; t (7) = 0.471, p = 0.652); D1R:Cre - ::sh Shank3 mice (i; t (6) = 0.264, p = 0.801) and D1R:Cre + ::sh Shank3 mice (j; t (8) = 3.443, p = 0.009). Right: juvenile preference index (one-sample t-tests against chance level = 0.5: D1R:Cre - ::scr Shank3 ; t (7) = 6.395, p < 0.001, D1R:Cre + ::scr Shank3 ; t (7) = 0.054, p = 0.958, D1R:Cre - ::sh Shank3 ; t (6) = 0.334, p = 0.750, D1R:Cre + ::sh Shank3 ; t (8) = 3.706, p = 0.006). Error bars report SEM.

Journal: bioRxiv

Article Title: Inhibition of TRPV4 rescues circuit and social deficits unmasked by acute inflammatory response in a Shank3 mouse model of Autism

doi: 10.1101/2021.10.13.464215

Figure Lengend Snippet: (a) Experimental design. Drd1a-dTomato mice were injected neonatally in the NAc with scr or sh Shank3 virus and whole-cell patch clamp recordings were performed during early adulthood. (a’) Representative images of the NAc of a D1R-tom+::sh Shank3 mouse (scale bar: 50 µm). (b) Example traces at 300 pA depolarizing current injection in D1R-tom+ MSNs infected with scr Shank3 (left) or with sh Shank3 (right). (c) Number of action potentials (nAPs) across increasing depolarizing current steps (0-500 pA) for D1R-tom+::scr Shank3 and sh Shank3 MSNs, in presence of picrotoxin and kynurenic acid. (Repeated measures ANOVA, virus main effect F (1,14) = 10.88, p = 0.005, current steps main effect F (10, 140) = 7.727, p < 0.001, scr Shank3 n = 8 cells, 3 mice, sh Shank3 n = 8 cells, 3 mice). (d) Example traces at 300 pA depolarizing current injection in D1R-tom-MSNs infected with scr Shank3 (left) or with sh Shank3 (right). (e) Number of action potential (nAPs) across increasing depolarizing current steps (0-500 pA) for D1R-tom-::scr Shank3 and sh Shank3 MSNs, in presence of picrotoxin and kynurenic acid. (Repeated measure (RM) two-way ANOVA, virus main effect F (1,18) = 0.098, p = 0.758, current steps main effect F (10 , 180) = 14.58, p < 0.001, n = 8 cells, 3 mice (sh), n=12 cells, 3 mice (scr)). (f) Experimental design. D1R-Cre positive (D1R:Cre + ) and negative (D1R:Cre - ) mice were injected neonatally in the NAc with scr or sh Shank3 virus and after P30 with AAV-hSyn-DIO-hM4Di-mCherry (DREADD). After 4 weeks, allowing for virus expression, the mice underwent social behaviour assessment in the three-chamber task. Mice were intraperitoneally injected with CNO 30 min before starting the test. (f’) Representative images of the NAc of a D1R:Cre + mouse infected with sh Shank3 and DREADD viruses (scale bar: 50 µm). (g, h, i, j ) Left: time around the target during the social preference test for D1R:Cre - ::scr Shank3 mice (g; t (7) = 5.453, p = 0.001); D1R:Cre + ::scr Shank3 mice (h; t (7) = 0.471, p = 0.652); D1R:Cre - ::sh Shank3 mice (i; t (6) = 0.264, p = 0.801) and D1R:Cre + ::sh Shank3 mice (j; t (8) = 3.443, p = 0.009). Right: juvenile preference index (one-sample t-tests against chance level = 0.5: D1R:Cre - ::scr Shank3 ; t (7) = 6.395, p < 0.001, D1R:Cre + ::scr Shank3 ; t (7) = 0.054, p = 0.958, D1R:Cre - ::sh Shank3 ; t (6) = 0.334, p = 0.750, D1R:Cre + ::sh Shank3 ; t (8) = 3.706, p = 0.006). Error bars report SEM.

Article Snippet: Viruses used in this study: (1) purified scr Shank3 and sh Shank3 (AAV1-GFP-U6-scrmbshRNA; titer: 5.9×10 13 GC/mL and AAV5-ZacF-U6-luczsGreen-sh Shank3 ; titer: 7.4×10 13 GC/mL, VectorBioLab); (2) AAV5/hsyn-DIO-hM4D(Gi)-mCherry (AV44961, titer: 5.5×10 12 virus molecules/mL, UNC GTC vector core).

Techniques: Injection, Virus, Patch Clamp, Infection, Expressing

(a) Experimental design. Drd1a-dTomato mice were injected neonatally in the NAc with scr or sh Shank3 virus. At P30 NAc was dissected and FACsorted in 4 different cell populations (D1R-tom+, D1R-tom-, D1R-tom+::AAV and D1R-tom-::AAV). For each cell population we carried out bulk RNA sequencing. (b) Worst-case scenario selected altered genes in scr vs sh testing clearly discriminated infected cells, both D1R+ and D1R- in PCA analysis. (c) While non-infected samples do not share common genes significantly altered in scr vs sh testing, infected D1R+ and D1R- share a core set of 68 altered genes. (d) Overall GO:Term analysis of infected D1R+ significantly altered genes highlights the relevance of inflammatory mechanisms, as well as cell adhesion-, localization- and movement-related functions. (e) D1R-tom+ altered genes include genes expressing proteins directly involved in electrophysiological properties, including the Transient receptor potential vanilloid 4 ( Trpv4 ). (f) Experimental design. Drd1a-dTomato mice were injected neonatally in the NAc with scr or sh Shank3 virus and whole-cell patch clamp recordings were performed during early adulthood. (g) Right: example traces from 300 pA depolarizing current injection in D1R+ MSNs infected with scr Shank3 treated with vehicle (up), D1R+ MSNs infected with sh Shank3 treated with vehicle (middle) and D1R+ MSNs infected with sh Shank3 treated with HC-067047 (down). Left: number of action potentials (nAPs) across increasing depolarizing current steps (0-500 pA) for D1R-tom+::scr Shank3 and sh Shank3 MSNs in the presence of Trpv4 antagonist (HC-067047). (Repeated measures ANOVA, drug main effect F (2, 31) = 5.883, p = 0.007, current steps main effect F (10 , 310) = 24.15, p < 0.001, drug by current steps interaction F (20 , 310) = 1.685, p = 0.035 n = 12 cells, 4 mice (sh Shank3 -Veh), n=10 cells, 3 mice (sh Shank3 -Trpv4), n = 12 cells, 4 mice (scr Shank3 -Veh)). (h) Experimental design. C57BL6/j mice were injected neonatally in the NAc with scr or sh Shank3 virus and at P50-60 were bilaterally cannulated above the NAc. After 7 days, mice underwent the three-chamber social interaction assay. Scr Shank3 were infused with vehicle (aCSF/DMSO 0.3%). On the other hand, sh Shank3 mice were infused with either vehicle (aCSF/DMSO 0.3%) or HC-067047 (2µg in aCSF/DMSO 0.3%) 10 min before to start the test. (h’) Representative image of the injection site and cannula placement above the NAc (scale bar: 250 µm). (i, j, k) Left: time around the target during the social preference test for mice infected with scr Shank3 and infused with vehicle (i, t (5) = 6.304, p = 0.002), mice infected with sh Shank3 and infused with vehicle (j, t (7) = 0.869, p = 0.414) or with HC-067047 (k, t (7) = 4.324, p = 0.004). Right: juvenile preference index for mice infused either with vehicle or with HC-067047 (one-sample t-tests against chance level = 0.5: (f); t (5) = 6.459, p = 0.001, (g); t (7) = 1.02, p = 0.342, (h); t (7) = 6.078, p = 0.001). (l) Juvenile preference index comparison between sh Shank3 -vehicle and sh Shank3 -HC-067047 ( t (7) = 2.6, p = 0.035). Error bars report SEM.

Journal: bioRxiv

Article Title: Inhibition of TRPV4 rescues circuit and social deficits unmasked by acute inflammatory response in a Shank3 mouse model of Autism

doi: 10.1101/2021.10.13.464215

Figure Lengend Snippet: (a) Experimental design. Drd1a-dTomato mice were injected neonatally in the NAc with scr or sh Shank3 virus. At P30 NAc was dissected and FACsorted in 4 different cell populations (D1R-tom+, D1R-tom-, D1R-tom+::AAV and D1R-tom-::AAV). For each cell population we carried out bulk RNA sequencing. (b) Worst-case scenario selected altered genes in scr vs sh testing clearly discriminated infected cells, both D1R+ and D1R- in PCA analysis. (c) While non-infected samples do not share common genes significantly altered in scr vs sh testing, infected D1R+ and D1R- share a core set of 68 altered genes. (d) Overall GO:Term analysis of infected D1R+ significantly altered genes highlights the relevance of inflammatory mechanisms, as well as cell adhesion-, localization- and movement-related functions. (e) D1R-tom+ altered genes include genes expressing proteins directly involved in electrophysiological properties, including the Transient receptor potential vanilloid 4 ( Trpv4 ). (f) Experimental design. Drd1a-dTomato mice were injected neonatally in the NAc with scr or sh Shank3 virus and whole-cell patch clamp recordings were performed during early adulthood. (g) Right: example traces from 300 pA depolarizing current injection in D1R+ MSNs infected with scr Shank3 treated with vehicle (up), D1R+ MSNs infected with sh Shank3 treated with vehicle (middle) and D1R+ MSNs infected with sh Shank3 treated with HC-067047 (down). Left: number of action potentials (nAPs) across increasing depolarizing current steps (0-500 pA) for D1R-tom+::scr Shank3 and sh Shank3 MSNs in the presence of Trpv4 antagonist (HC-067047). (Repeated measures ANOVA, drug main effect F (2, 31) = 5.883, p = 0.007, current steps main effect F (10 , 310) = 24.15, p < 0.001, drug by current steps interaction F (20 , 310) = 1.685, p = 0.035 n = 12 cells, 4 mice (sh Shank3 -Veh), n=10 cells, 3 mice (sh Shank3 -Trpv4), n = 12 cells, 4 mice (scr Shank3 -Veh)). (h) Experimental design. C57BL6/j mice were injected neonatally in the NAc with scr or sh Shank3 virus and at P50-60 were bilaterally cannulated above the NAc. After 7 days, mice underwent the three-chamber social interaction assay. Scr Shank3 were infused with vehicle (aCSF/DMSO 0.3%). On the other hand, sh Shank3 mice were infused with either vehicle (aCSF/DMSO 0.3%) or HC-067047 (2µg in aCSF/DMSO 0.3%) 10 min before to start the test. (h’) Representative image of the injection site and cannula placement above the NAc (scale bar: 250 µm). (i, j, k) Left: time around the target during the social preference test for mice infected with scr Shank3 and infused with vehicle (i, t (5) = 6.304, p = 0.002), mice infected with sh Shank3 and infused with vehicle (j, t (7) = 0.869, p = 0.414) or with HC-067047 (k, t (7) = 4.324, p = 0.004). Right: juvenile preference index for mice infused either with vehicle or with HC-067047 (one-sample t-tests against chance level = 0.5: (f); t (5) = 6.459, p = 0.001, (g); t (7) = 1.02, p = 0.342, (h); t (7) = 6.078, p = 0.001). (l) Juvenile preference index comparison between sh Shank3 -vehicle and sh Shank3 -HC-067047 ( t (7) = 2.6, p = 0.035). Error bars report SEM.

Article Snippet: Viruses used in this study: (1) purified scr Shank3 and sh Shank3 (AAV1-GFP-U6-scrmbshRNA; titer: 5.9×10 13 GC/mL and AAV5-ZacF-U6-luczsGreen-sh Shank3 ; titer: 7.4×10 13 GC/mL, VectorBioLab); (2) AAV5/hsyn-DIO-hM4D(Gi)-mCherry (AV44961, titer: 5.5×10 12 virus molecules/mL, UNC GTC vector core).

Techniques: Injection, Virus, RNA Sequencing, Infection, Expressing, Patch Clamp, Comparison

(a) Differential expression analysis of AAV-scrShank3 vs AAV-ShShank3 shows small indirect transcriptional effect in non-infected samples, while infected samples display the stronger transcriptomic alterations. In both D1R+ (b) and D1R- (c) SFARI associated genes are altered, supporting the link between Shank3 downregulation with an autism-related phenotype.

Journal: bioRxiv

Article Title: Inhibition of TRPV4 rescues circuit and social deficits unmasked by acute inflammatory response in a Shank3 mouse model of Autism

doi: 10.1101/2021.10.13.464215

Figure Lengend Snippet: (a) Differential expression analysis of AAV-scrShank3 vs AAV-ShShank3 shows small indirect transcriptional effect in non-infected samples, while infected samples display the stronger transcriptomic alterations. In both D1R+ (b) and D1R- (c) SFARI associated genes are altered, supporting the link between Shank3 downregulation with an autism-related phenotype.

Article Snippet: Viruses used in this study: (1) purified scr Shank3 and sh Shank3 (AAV1-GFP-U6-scrmbshRNA; titer: 5.9×10 13 GC/mL and AAV5-ZacF-U6-luczsGreen-sh Shank3 ; titer: 7.4×10 13 GC/mL, VectorBioLab); (2) AAV5/hsyn-DIO-hM4D(Gi)-mCherry (AV44961, titer: 5.5×10 12 virus molecules/mL, UNC GTC vector core).

Techniques: Quantitative Proteomics, Infection

(a) Behavioural task paradigm. (b, c) Left: Time spent around the target during social preference test for Shank3 +/+ and Shank3 +/- mice (paired-samples t-tests for object- vs. social: Shank3 +/+ : t (9) = 5.167, p < 0.001; Shank3 +/- : t (12) = 3.026, p = 0.011). Right: juvenile preference index (one-sample t-tests against chance level = 0.5: Shank3 +/+ : t (9) = 5.617, p < 0.001; Shank3 +/- : t (12) = 3.146, p = 0.008). (d) Time spent in the juvenile, object or in center chamber during social preference test for Shank3 +/+ , Shank3 +/- and Shank3 -/- mice (Paired-samples t-tests for object- vs. social-containing chambers: Shank3 +/+ : t (9) = 3.269, p = 0.009; Shank3 +/- : t (12) = 2.705, p = 0.019). (e) Distance moved during social preference test (Unpaired-samples t-tests: t (21) = 0.9307, p = 0.363).

Journal: bioRxiv

Article Title: Inhibition of TRPV4 rescues circuit and social deficits unmasked by acute inflammatory response in a Shank3 mouse model of Autism

doi: 10.1101/2021.10.13.464215

Figure Lengend Snippet: (a) Behavioural task paradigm. (b, c) Left: Time spent around the target during social preference test for Shank3 +/+ and Shank3 +/- mice (paired-samples t-tests for object- vs. social: Shank3 +/+ : t (9) = 5.167, p < 0.001; Shank3 +/- : t (12) = 3.026, p = 0.011). Right: juvenile preference index (one-sample t-tests against chance level = 0.5: Shank3 +/+ : t (9) = 5.617, p < 0.001; Shank3 +/- : t (12) = 3.146, p = 0.008). (d) Time spent in the juvenile, object or in center chamber during social preference test for Shank3 +/+ , Shank3 +/- and Shank3 -/- mice (Paired-samples t-tests for object- vs. social-containing chambers: Shank3 +/+ : t (9) = 3.269, p = 0.009; Shank3 +/- : t (12) = 2.705, p = 0.019). (e) Distance moved during social preference test (Unpaired-samples t-tests: t (21) = 0.9307, p = 0.363).

Article Snippet: Viruses used in this study: (1) purified scr Shank3 and sh Shank3 (AAV1-GFP-U6-scrmbshRNA; titer: 5.9×10 13 GC/mL and AAV5-ZacF-U6-luczsGreen-sh Shank3 ; titer: 7.4×10 13 GC/mL, VectorBioLab); (2) AAV5/hsyn-DIO-hM4D(Gi)-mCherry (AV44961, titer: 5.5×10 12 virus molecules/mL, UNC GTC vector core).

Techniques:

(a) Experimental design. Shank3 +/+ and Shank3 +/- were intraperitoneally injected with LPS or vehicle and 24 hrs later they were subjected to 3-chamber task. (b, c, d, e) Left: Time spent around the target (Paired-samples t-tests for object- vs. social: (b); t (7) = 7.686, p < 0.001, (c); t (8) = 4.199, p = 0.003, (d); t (8) = 3.462, p = 0.009, (e); t (9) = 0.935, p = 0.374). Right: juvenile preference index (one-sample t-tests against chance level = 0.5: (b); t (7) = 7.2, p < 0.001, (c); t (8) = 5.262, p < 0.001, (d); t (8) = 3.734, p = 0.006, (e); t (9) = 0.9747, p = 0.355). (f) Experimental design. Shank3 +/+ and Shank3 +/- were intraperitoneally injected with LPS and 7 days later were subjected to 3-chamber task. (g, h) Left: Time spent around the target (Paired-samples t-tests for object- vs. social: (g); t (6) = 5.979, p = 0.001, (h); t (9) = 2.759, p = 0.022). Right: juvenile preference index (one-sample t-tests against chance level = 0.5: (g); t (6) = 6.054, p < 0.001, (h); t (9) = 2.463, p = 0.036). (i) mRNA expression analysis of IL-1β , TNF-α and Trpv4 genes after LPS challenge in Shank3 +/+ (IL-1 β one way ANOVA followed by Sidak’s multiple comparisons test, F (2,9) =10.33, p=0.005 ; TNF-α Kruskal-Wallis statistic 7.538, p=0.012; Trpv4 one way ANOVA followed by Sidak’s multiple comparisons test, F (2,9) =2.768, p = 0.116 ). (j) mRNA expression analysis of IL-1β , TNF-α and Trpv4 genes after LPS challenge in Shank3 +/- (IL-1β Kruskal-Wallis statistic 9.002, p=0.002 ; TNF-α one way ANOVA followed by Sidak’s multiple comparisons test, F (2.10) =10.27, p=0.004; Trpv4 one way ANOVA followed by Sidak’s multiple comparisons test: F (2,9) = 31.26, p < 0.001 ). (k) Experimental design. Shank3 +/- were crossed with Drd1a-tdTomato mice labelling specifically D1R-MSNs in Shank3 +/- background. Ex-vivo patch clamp recordings were made 24 hrs after the LPS injection. (l) Whole-cell recording of Trpv4 current after LPS challenge in Shank3 +/+- and Shank3 +/- mice (Repeated measures ANOVA, voltage steps main effect F (1.974,43.29) = 16.15, p=0.001 , genotype by voltage steps interaction F (120 , 880) = 1.451, p = 0.002; n = 5 cells, 2 mice ( Shank3 +/+ ), n=5 cells, 2 mice ( Shank3 +/+ +LPS), n = 7 cells, 2 mice ( Shank3 -/+ ) n = 9 cells, 2 mice ( Shank3 -/+ +LPS)) (m) Example traces from 300 pA depolarizing current injection in D1R+ MSNs of Shank3 +/- mice after vehicle IP injection and treated with vehicle (left), D1R+ MSNs of Shank3 +/- mice after LPS challenge and treated with vehicle (middle), and in D1R+ MSNs of Shank3 +/- mice after LPS challenge and treated with HC-067047. (n) Number of action potentials (nAPs) across increasing depolarizing current steps (0-500 pA) for D1R-tom+:: Shank3 +/- MSNs after LPS challenge (Repeated measures ANOVA, LPS challenge main effect F (2.30) = 3.034, p = 0.063, current steps main effect F (10 , 300) = 28.08, p < 0.001, LPS challenge by current steps interaction F (20 , 300) = 2.042, p = 0.006, n = 10 cells, 4 mice (D1R-tom+:: Shank3 +/- + veh), n = 13 cells, 3 mice (D1R-tom+:: Shank3 +/- + LPS), n = 10 cells, 3 mice (D1R-tom+:: Shank3 +/- + LPS + HC067047)). Error bars report SEM.

Journal: bioRxiv

Article Title: Inhibition of TRPV4 rescues circuit and social deficits unmasked by acute inflammatory response in a Shank3 mouse model of Autism

doi: 10.1101/2021.10.13.464215

Figure Lengend Snippet: (a) Experimental design. Shank3 +/+ and Shank3 +/- were intraperitoneally injected with LPS or vehicle and 24 hrs later they were subjected to 3-chamber task. (b, c, d, e) Left: Time spent around the target (Paired-samples t-tests for object- vs. social: (b); t (7) = 7.686, p < 0.001, (c); t (8) = 4.199, p = 0.003, (d); t (8) = 3.462, p = 0.009, (e); t (9) = 0.935, p = 0.374). Right: juvenile preference index (one-sample t-tests against chance level = 0.5: (b); t (7) = 7.2, p < 0.001, (c); t (8) = 5.262, p < 0.001, (d); t (8) = 3.734, p = 0.006, (e); t (9) = 0.9747, p = 0.355). (f) Experimental design. Shank3 +/+ and Shank3 +/- were intraperitoneally injected with LPS and 7 days later were subjected to 3-chamber task. (g, h) Left: Time spent around the target (Paired-samples t-tests for object- vs. social: (g); t (6) = 5.979, p = 0.001, (h); t (9) = 2.759, p = 0.022). Right: juvenile preference index (one-sample t-tests against chance level = 0.5: (g); t (6) = 6.054, p < 0.001, (h); t (9) = 2.463, p = 0.036). (i) mRNA expression analysis of IL-1β , TNF-α and Trpv4 genes after LPS challenge in Shank3 +/+ (IL-1 β one way ANOVA followed by Sidak’s multiple comparisons test, F (2,9) =10.33, p=0.005 ; TNF-α Kruskal-Wallis statistic 7.538, p=0.012; Trpv4 one way ANOVA followed by Sidak’s multiple comparisons test, F (2,9) =2.768, p = 0.116 ). (j) mRNA expression analysis of IL-1β , TNF-α and Trpv4 genes after LPS challenge in Shank3 +/- (IL-1β Kruskal-Wallis statistic 9.002, p=0.002 ; TNF-α one way ANOVA followed by Sidak’s multiple comparisons test, F (2.10) =10.27, p=0.004; Trpv4 one way ANOVA followed by Sidak’s multiple comparisons test: F (2,9) = 31.26, p < 0.001 ). (k) Experimental design. Shank3 +/- were crossed with Drd1a-tdTomato mice labelling specifically D1R-MSNs in Shank3 +/- background. Ex-vivo patch clamp recordings were made 24 hrs after the LPS injection. (l) Whole-cell recording of Trpv4 current after LPS challenge in Shank3 +/+- and Shank3 +/- mice (Repeated measures ANOVA, voltage steps main effect F (1.974,43.29) = 16.15, p=0.001 , genotype by voltage steps interaction F (120 , 880) = 1.451, p = 0.002; n = 5 cells, 2 mice ( Shank3 +/+ ), n=5 cells, 2 mice ( Shank3 +/+ +LPS), n = 7 cells, 2 mice ( Shank3 -/+ ) n = 9 cells, 2 mice ( Shank3 -/+ +LPS)) (m) Example traces from 300 pA depolarizing current injection in D1R+ MSNs of Shank3 +/- mice after vehicle IP injection and treated with vehicle (left), D1R+ MSNs of Shank3 +/- mice after LPS challenge and treated with vehicle (middle), and in D1R+ MSNs of Shank3 +/- mice after LPS challenge and treated with HC-067047. (n) Number of action potentials (nAPs) across increasing depolarizing current steps (0-500 pA) for D1R-tom+:: Shank3 +/- MSNs after LPS challenge (Repeated measures ANOVA, LPS challenge main effect F (2.30) = 3.034, p = 0.063, current steps main effect F (10 , 300) = 28.08, p < 0.001, LPS challenge by current steps interaction F (20 , 300) = 2.042, p = 0.006, n = 10 cells, 4 mice (D1R-tom+:: Shank3 +/- + veh), n = 13 cells, 3 mice (D1R-tom+:: Shank3 +/- + LPS), n = 10 cells, 3 mice (D1R-tom+:: Shank3 +/- + LPS + HC067047)). Error bars report SEM.

Article Snippet: Viruses used in this study: (1) purified scr Shank3 and sh Shank3 (AAV1-GFP-U6-scrmbshRNA; titer: 5.9×10 13 GC/mL and AAV5-ZacF-U6-luczsGreen-sh Shank3 ; titer: 7.4×10 13 GC/mL, VectorBioLab); (2) AAV5/hsyn-DIO-hM4D(Gi)-mCherry (AV44961, titer: 5.5×10 12 virus molecules/mL, UNC GTC vector core).

Techniques: Injection, Expressing, Ex Vivo, Patch Clamp

LPS challenge induces social deficits in Shank3 +/- mice after 24 hours: (a) Time spent in the juvenile, object or in the center chamber during social preference test for Shank3 +/+ and Shank3 +/- previously injected with vehicle or LPS (Paired-samples t-tests for object- vs. social-containing chambers: Shank3 +/+ + veh: t (7) = 4.838, p = 0.002, Shank3 +/+ + LPS: t (8) = 3.87, p = 0.005, Shank3 +/- + veh: t (8) = 4.526, p = 0.002, Shank3 +/- + LPS: t (9) = 0.3939, p = 0.703). (b) Distance moved during social preference test (two-way ANOVA followed by Bonferroni’s multiple comparisons test: LPS treatment main effect F (1,32) =58.03, p<0.001 ). 7 days after LPS challenge the sociability and the distance moved are not impaired anymore: (c) Time spent in the juvenile, object or in the center chamber during social preference test for Shank3 +/+ and Shank3 +/- previously injected LPS (Paired-samples t-tests for object- vs. social-containing chambers: Shank3 +/+ + LPS: t (7) = 5.083, p = 0.002, Shank3 +/- + LPS: t (10) = 2.536, p = 0.032. (d) Distance moved during social preference test (unpaired t-test t (15) =0.3116, p=0.76). Error bars report SEM.

Journal: bioRxiv

Article Title: Inhibition of TRPV4 rescues circuit and social deficits unmasked by acute inflammatory response in a Shank3 mouse model of Autism

doi: 10.1101/2021.10.13.464215

Figure Lengend Snippet: LPS challenge induces social deficits in Shank3 +/- mice after 24 hours: (a) Time spent in the juvenile, object or in the center chamber during social preference test for Shank3 +/+ and Shank3 +/- previously injected with vehicle or LPS (Paired-samples t-tests for object- vs. social-containing chambers: Shank3 +/+ + veh: t (7) = 4.838, p = 0.002, Shank3 +/+ + LPS: t (8) = 3.87, p = 0.005, Shank3 +/- + veh: t (8) = 4.526, p = 0.002, Shank3 +/- + LPS: t (9) = 0.3939, p = 0.703). (b) Distance moved during social preference test (two-way ANOVA followed by Bonferroni’s multiple comparisons test: LPS treatment main effect F (1,32) =58.03, p<0.001 ). 7 days after LPS challenge the sociability and the distance moved are not impaired anymore: (c) Time spent in the juvenile, object or in the center chamber during social preference test for Shank3 +/+ and Shank3 +/- previously injected LPS (Paired-samples t-tests for object- vs. social-containing chambers: Shank3 +/+ + LPS: t (7) = 5.083, p = 0.002, Shank3 +/- + LPS: t (10) = 2.536, p = 0.032. (d) Distance moved during social preference test (unpaired t-test t (15) =0.3116, p=0.76). Error bars report SEM.

Article Snippet: Viruses used in this study: (1) purified scr Shank3 and sh Shank3 (AAV1-GFP-U6-scrmbshRNA; titer: 5.9×10 13 GC/mL and AAV5-ZacF-U6-luczsGreen-sh Shank3 ; titer: 7.4×10 13 GC/mL, VectorBioLab); (2) AAV5/hsyn-DIO-hM4D(Gi)-mCherry (AV44961, titer: 5.5×10 12 virus molecules/mL, UNC GTC vector core).

Techniques: Injection

(a) Experimental design. Adult Shank3 +/- mice were intraperitoneally injected with LPS and 24 hours later were subjected to the behavioural task. 30 minutes before the test, mice were infused (in the NAc) either with Trpv4 antagonist (HC-067047) or vehicle. (b, c) Left: Time spent around the target for Shank3 +/ mice after LPS challenge and vehicle or HC-067047 infusion in the NAc (Paired-samples t-tests for object- vs. social: (b); t (6) = 0.408, p = 0.687, (c); t (6) = 2.787, p = 0.032). Right: juvenile preference index (one-sample t-tests against chance level = 0.5: (b); t (6) = 0.629, p = 0.4388, (c); t (6) = 2.852, p = 0.029). Error bars report SEM.

Journal: bioRxiv

Article Title: Inhibition of TRPV4 rescues circuit and social deficits unmasked by acute inflammatory response in a Shank3 mouse model of Autism

doi: 10.1101/2021.10.13.464215

Figure Lengend Snippet: (a) Experimental design. Adult Shank3 +/- mice were intraperitoneally injected with LPS and 24 hours later were subjected to the behavioural task. 30 minutes before the test, mice were infused (in the NAc) either with Trpv4 antagonist (HC-067047) or vehicle. (b, c) Left: Time spent around the target for Shank3 +/ mice after LPS challenge and vehicle or HC-067047 infusion in the NAc (Paired-samples t-tests for object- vs. social: (b); t (6) = 0.408, p = 0.687, (c); t (6) = 2.787, p = 0.032). Right: juvenile preference index (one-sample t-tests against chance level = 0.5: (b); t (6) = 0.629, p = 0.4388, (c); t (6) = 2.852, p = 0.029). Error bars report SEM.

Article Snippet: Viruses used in this study: (1) purified scr Shank3 and sh Shank3 (AAV1-GFP-U6-scrmbshRNA; titer: 5.9×10 13 GC/mL and AAV5-ZacF-U6-luczsGreen-sh Shank3 ; titer: 7.4×10 13 GC/mL, VectorBioLab); (2) AAV5/hsyn-DIO-hM4D(Gi)-mCherry (AV44961, titer: 5.5×10 12 virus molecules/mL, UNC GTC vector core).

Techniques: Injection

(a) Time spent in compartments for Shank3 +/- mice after LPS challenge and with vehicle or HC-067047 infusion in the NAc (Paired-samples t-tests for object- vs. social-containing chambers: Shank3 +/- + veh: t (6) = 1.3229, p = 0.232, Shank3 +/- + HC-067047: t (6) = 1.17, p = 0.286). (b) Distance moved during social preference test (unpaired t-test: t (12) = 0.3708, p = 0.717). Error bars report SEM.

Journal: bioRxiv

Article Title: Inhibition of TRPV4 rescues circuit and social deficits unmasked by acute inflammatory response in a Shank3 mouse model of Autism

doi: 10.1101/2021.10.13.464215

Figure Lengend Snippet: (a) Time spent in compartments for Shank3 +/- mice after LPS challenge and with vehicle or HC-067047 infusion in the NAc (Paired-samples t-tests for object- vs. social-containing chambers: Shank3 +/- + veh: t (6) = 1.3229, p = 0.232, Shank3 +/- + HC-067047: t (6) = 1.17, p = 0.286). (b) Distance moved during social preference test (unpaired t-test: t (12) = 0.3708, p = 0.717). Error bars report SEM.

Article Snippet: Viruses used in this study: (1) purified scr Shank3 and sh Shank3 (AAV1-GFP-U6-scrmbshRNA; titer: 5.9×10 13 GC/mL and AAV5-ZacF-U6-luczsGreen-sh Shank3 ; titer: 7.4×10 13 GC/mL, VectorBioLab); (2) AAV5/hsyn-DIO-hM4D(Gi)-mCherry (AV44961, titer: 5.5×10 12 virus molecules/mL, UNC GTC vector core).

Techniques: