mouse anti trio  (Developmental Studies Hybridoma Bank)

Journal: Scientific Reports

Article Title: Elevated serotonin receptor 2A signaling restores learning and memory in a Fragile X syndrome model

doi: 10.1038/s41598-025-34492-4

Figure Lengend Snippet: Learning and memory regulation via Mushroom Body serotonin signaling. ( A ) Aversive olfactory conditioning apparatus used for Pavlovian learning/memory assays. The setup includes red light illumination, humidity control, vacuum airflow odorant delivery, copper-grid shock tube with electrical stimulator (training), a vertical elevator for animal delivery, and a T-maze with two choice tubes (testing). Animals are trained with odorant cues paired to electric shock in the shock tube, and then scored in the T-maze test tubes. ( B ) Schematic flowchart of the learning and memory assays. The conditioning consists of alternating exposures to CS + odor paired with 12 shocks (80 V, 1.5 s duration, 5 s interval), air interval, and then the CS- odor without shock, followed by either immediate or delayed (24 h) T-maze testing. Across independent trials, OCT and MCH were assigned as CS⁺ and CS⁻ in alternative experiments. Performance index (PI = (CS - – CS + )/(CS - + CS + )) reports learning/memory abilities. ( C ) Quantification of learning (left) and memory (right) PIs in 5 genotypes shown: genetic background control ( w 1118 ); global UH1 -Gal4 serotonin transporter (SERT) and serotonin receptor 2A (5HT 2A R) RNAi; FXS disease model ( dfmr1 null mutant) alone and with UH1 -Gal4/ + ( dfmr1 control). Data show individual trials (n = 16/genotype), mean ± SEM, and one-way ANOVA with Tukey’s multiple comparisons tests. For learning (left), there is a significant effect of genotype (F(4,75) = 16.54), with no significant difference between w 1118 and SERT RNAi ( p = 0.9999), while 5HT 2A R RNAi ( p = 2.588 × 10 –4 ) and dfmr1 null mutants ( p = 1.924 × 10 –6 ) show reduced performance, with the dfmr1 transgenic control not significantly different ( p = 0.9999). For memory, there is a significant effect of genotype (F(4, 75) = 36.21), with no significant difference between w 1118 and SERT RNAi ( p = 0.9759), whereas 5HT 2A R RNAi ( p = 6.347 × 10 –3 ) and dfmr1 null mutants ( p = 3.0 × 10 –11 ) show decreased performance. Again, dfmr1 null and dfmr1 control are not significantly different ( p = 0.9999). The significance is indicated as p > 0.05 (not significant; ns), p < 0.01 (**), p < 0.001 ( ***), and p < 0.0001 (****). ( D ) Drosophila brain confocal images labeled with anti-Trio to reveal the Mushroom Body. Left : Whole-brain at 40 × magnification shows Mushroom Body (MB) and optic lobes (OL). Right : Higher magnification at 63 × showing MB α′, β′, and γ lobes. Fluorescence intensity is shown on a 16-color LUT scale. ( E ) Serotonergic innervation of the MB in an anatomical reconstruction from FlyWire codex. Dorsal paired medial neurons (DPM, green) provide serotonergic input onto MB Kenyon cells (KC, magenta; small subset shown for clarity).

Article Snippet: Primary antibodies used: rabbit anti-5-HT (Immunostar, 20,080 1:1,000), rabbit anti-5HT 2A R (Abcam, ab140524, 1:100), and mouse anti-Trio (Developmental Studies Hybridoma Bank (DSHB), 9.4A, 1:100).

Techniques: Olfactory, Control, Mutagenesis, Transgenic Assay, Labeling, Fluorescence

Journal: Scientific Reports

Article Title: Elevated serotonin receptor 2A signaling restores learning and memory in a Fragile X syndrome model

doi: 10.1038/s41598-025-34492-4

Figure Lengend Snippet: Tryptophan hydroxylase overexpression rectifies FXS learning/memory. Performance index quantification of learning ( A ) and memory ( B ) across five genotypes; genetic background control ( w 1118 ; green), FXS model ( dfmr1 ; red), and UAS- tryptophan hydroxylase ( Trhn ) overexpression in the dfmr1 background driven by UH1- (ubiquitous), elav- (neuronal), and repo- (glial) Gal4 lines (blue). Data show individual trials (n = 16 each), mean ± SEM, and one-way ANOVA with Tukey’s multiple comparisons tests. Learning performance shows a significant effect of genotype (F(4,75) = 17.93). For learning, a significant impairment occurs between w 1118 and dfmr1 ( p = 4.662 × 10 –7 ), and a significant improvement between dfmr1 alone and with UH1- Gal4 driven UAS- Trhn ( p = 3.367 × 10 –9 ). No significant differences occur in any other comparisons. Memory performance also shows a significant effect of genotype (F(4,75) = 10.58). For memory trials, a significant impairment occurs between w 1118 and dfmr1 ( p = 3.902 × 10 –6 ), and significant improvement between dfmr1 alone and with UH1- Gal4 driven UAS- Trhn ( p = 2.645 × 10 –6 ). No significant differences occur in any other comparisons. ( C ) Representative Mushroom Body lobe (anti-Trio, green outline) anti-serotonin (5-HT, grey scale) labeling in the same five above genotypes in untrained (top row) and trained (bottom row) conditions. Quantification of MB 5-HT fluorescence intensity from untrained ( D ) and T-maze trained ( E ) conditions. Data show individual brains (n = 10–15/condition), mean ± SEM, and one-way ANOVA with Tukey’s multiple comparisons tests. For the untrained basal condition, the overall effect of genotype is significant (F(4, 47) = 28.50), with a significant elevation occurring between dfmr1 nulls alone versus with UH1- Gal4 driven UAS- Trhn ( p = 1.477 × 10 –8 ). No significant differences occur in any other comparisons. For the T-maze trained condition, the overall effect of genotype is also significant (F(4, 45) = 25.75), with a significant increase between dfmr1 nulls alone versus with UH1- Gal4 driven UAS- Trhn ( p = 2.207 × 10 –7 ). No significant differences occur in any other comparisons. Significance is indicated as p > 0.05 (not significant; ns) and p < 0.0001 (****).

Article Snippet: Primary antibodies used: rabbit anti-5-HT (Immunostar, 20,080 1:1,000), rabbit anti-5HT 2A R (Abcam, ab140524, 1:100), and mouse anti-Trio (Developmental Studies Hybridoma Bank (DSHB), 9.4A, 1:100).

Techniques: Over Expression, Control, Labeling, Fluorescence

Journal: Scientific Reports

Article Title: Elevated serotonin receptor 2A signaling restores learning and memory in a Fragile X syndrome model

doi: 10.1038/s41598-025-34492-4

Figure Lengend Snippet: Serotonin transporter (SERT) knockdown restores FXS learning/memory. Learning ( A ) and memory ( B ) performance index quantifications across five genotypes; genetic background control ( w 1118 ; green), FXS model ( dfmr1 ; red), and UAS- serotonin transporter ( SERT ) RNAi in the dfmr1 mutant background driven by UH1- (ubiquitous), elav- (neuronal), and repo- (glial) Gal4 lines (blue). Data show individual trials (n = 16 each), mean ± SEM, and one-way ANOVA with Tukey’s multiple comparisons tests. Learning performance shows a significant effect of genotype (F(4,75) = 16.40). For learning, a significant impairment occurs between w 1118 and dfmr1 ( p = 8.993 × 10 –6 ), and a significant improvement between dfmr1 alone vs with UH1- Gal4 driven SERT RNAi ( p = 4.388 × 10 –7 ). No significant differences occur in any of the other comparisons. Memory performance also shows a significant effect of genotype (F(4,75) = 17.00). For memory, a significant impairment occurs between w 1118 and dfmr1 ( p = 6.184 × 10 –9 ), and significant improvement between dfmr1 alone and with UH1- Gal4 driven UAS- SERT RNAi ( p = 6.644 × 10 –7 ). No significant differences occur in any other comparisons. ( C ) Mushroom Body lobe outline (anti-Trio, green) and anti-serotonin (5-HT, grey scale) labeling in the same five genotypes in untrained (top row) and trained (bottom row) conditions. Quantification of the MB 5-HT fluorescence intensity in both the untrained ( D ) and trained ( E ) conditions. Data show individual brains (n = 10–15/condition), mean ± SEM, and one-way ANOVA with Tukey’s multiple comparisons tests. For the untrained condition, the overall effect of genotype is significant (F(4, 45) = 15.62), with a significant elevation occurs between dfmr1 alone and with UH1- Gal4 driven UAS- SERT RNAi ( p = 1.285 × 10 –4 ). No significant differences occur in any of the other comparisons. For the trained condition, the overall effect of genotype is also significant (F(4, 48) = 12.19), with a significant increase likewise occurring between dfmr1 alone and with UH1- Gal4 driven UAS- SERT RNAi ( p = 1.267 × 10 –4 ). No significant differences occur in any other comparisons. Significance is indicated as p > 0.05 (not significant; ns), p < 0.001 (***), and p < 0.0001 (****).

Article Snippet: Primary antibodies used: rabbit anti-5-HT (Immunostar, 20,080 1:1,000), rabbit anti-5HT 2A R (Abcam, ab140524, 1:100), and mouse anti-Trio (Developmental Studies Hybridoma Bank (DSHB), 9.4A, 1:100).

Techniques: Knockdown, Control, Mutagenesis, Labeling, Fluorescence

Journal: Scientific Reports

Article Title: Elevated serotonin receptor 2A signaling restores learning and memory in a Fragile X syndrome model

doi: 10.1038/s41598-025-34492-4

Figure Lengend Snippet: 5HT 2A R knockdown only weakly exacerbates FXS model learning/memory. Performance index quantifications of learning ( A ) and memory ( B ) across five genotypes; the genetic background control ( w 1118 ; green), FXS model ( dfmr1 ; red), and UAS- 5HT 2A receptor ( 5HT 2A R ) RNAi in the dfmr1 null background driven by UH1- (ubiquitous), elav- (neuronal), and repo- (glial) Gal4 lines (blue). Data show all individual trials (n = 16 each), mean ± SEM, and one-way ANOVA with Tukey’s multiple comparisons tests. Learning performance shows a significant effect of genotype (F(4,75) = 21.02). For learning, significant differences occur between w 1118 and dfmr1 ( p = 7.460 × 10 –5 ), w 1118 and UH1 -Gal4 driven 5HT 2A R RNAi in the dfmr1 null mutant ( p = 4.617 × 10 –10 ), and between this condition and repo -Gal4 driven 5HT 2A R RNAi in the dfmr1 mutant ( p = 9.559 × 10 –3 ). A significant difference also occurs between dfmr1 and UH1 -Gal4 driven 5HT 2A R RNAi in the dfmr1 mutant ( p = 0.01548). No significant differences occur in the other comparisons. Memory performance also shows a significant effect of genotype (F(4,75) = 14.04). For memory, significant differences occur between w 1118 and dfmr1 ( p = 8.599 × 10 –5 ), and between w 1118 and UH1 -Gal4 driven 5HT 2A R RNAi in the dfmr1 mutant ( p = 7.094 × 10 –7 ). No other significant differences occur. ( C ) Representative Mushroom Body lobe (anti-Trio, green outline) anti-serotonin receptor 2A (5HT 2A R, grey scale) labeling in the same five genotypes in untrained (top row) and trained (bottom row) conditions. Quantification of MB 5-HT fluorescence intensity from untrained ( D ) and trained ( E ) conditions. Data show individual brains (n = 10–15 per condition), mean ± SEM, and one-way ANOVA with Tukey’s multiple comparisons tests. For the basal untrained condition, the overall effect of genotype is significant (F(4, 53) = 21.92), with a significant difference occurs between w 1118 and dfmr1 ( p = 7.858 × 10 –3 ), w 1118 and UH1 -Gal4 driven 5HT 2A R RNAi in the dfmr1 mutant ( p = 7.621 × 10 –9 ), and dfmr1 and UH1 -Gal4 driven 5HT 2A R RNAi in the dfmr1 mutant ( p = 5.732 × 10 –3 ). No significant differences occur in the other comparisons. For the trained condition, the overall effect of genotype is significant (F(4, 52) = 21.58), with a significant 5HT 2A R loss occurring between w 1118 and dfmr1 null ( p = 9.630 × 10 –3 ), w 1118 and UH1 -Gal4 driven 5HT 2A R RNAi in the dfmr1 null mutant ( p = 5.083 × 10 –9 ), and dfmr1 and UH1 -Gal4 driven 5HT 2A R RNAi in the dfmr1 null mutant ( p = 2.454 × 10 –3 ). No other significant differences occur. Significance is indicated as p > 0.05 (not significant, ns), p < 0.05 (*), p < 0.01 (**), and p < 0.0001 (****).

Article Snippet: Primary antibodies used: rabbit anti-5-HT (Immunostar, 20,080 1:1,000), rabbit anti-5HT 2A R (Abcam, ab140524, 1:100), and mouse anti-Trio (Developmental Studies Hybridoma Bank (DSHB), 9.4A, 1:100).

Techniques: Knockdown, Control, Mutagenesis, Labeling, Fluorescence

Journal: Scientific Reports

Article Title: Elevated serotonin receptor 2A signaling restores learning and memory in a Fragile X syndrome model

doi: 10.1038/s41598-025-34492-4

Figure Lengend Snippet: 5HT 2A R overexpression rectifies FXS model learning/memory performance. Learning ( A ) and memory ( B ) performance index quantifications across five genotypes; genetic background control ( w 1118 ; green), FXS model ( dfmr1 ; red), and UAS- 5HT 2A receptor ( 5HT 2A R ) overexpression in the dfmr1 background driven by UH1- (ubiquitous), elav- (neuronal), and repo- (glial) Gal4 lines (blue). Data show individual trials (n = 16 each), mean ± SEM, and one-way ANOVA with Tukey’s multiple comparisons tests. Learning performance shows a significant effect of genotype (F(4,75) = 21.30). For learning, a significant impairment occurs between w 1118 and dfmr1 ( p = 4.112 × 10 –7 ), improvement compared to dfmr1 with UH1 -Gal4 driven UAS- 5HT 2A R in the dfmr1 null ( p = 2.404 × 10 –5 ), and decrease from the UH1- to repo -Gal4 conditions ( p = 8.799 × 10 –3 ). Memory performance also shows a significant effect of genotype (F(4,75) = 7.881). For memory, a significant loss again occurs between w 1118 and dfmr1 ( p = 2.133 × 10 –5 ), which is restored comparing dfmr1 to UH1 -Gal4 driven UAS- 5HT 2A R in the dfmr1 null ( p = 2.762 × 10 –4 ). No significant differences occur in the other comparisons. ( C ) Mushroom Body lobe (anti-Trio, green outline) anti-serotonin receptor 2A (5HT 2A R, grey scale) labeling in the same five genotypes in the untrained (top row) and trained (bottom row) conditions. Quantification of the MB 5-HT 2A R fluorescence intensity in the untrained ( D ) and trained ( E ) conditions. Data show individual brains (n = 10–15 per condition), mean ± SEM, and one-way ANOVA with Tukey’s multiple comparisons tests. For the untrained condition, the overall effect of genotype is significant (F(4, 51) = 16.85), with a significant decrease occurs between w 1118 and dfmr1 ( p = 3.495 × 10 –3 ), w 1118 and UH1 -Gal4 driven UAS- 5HT 2A R in dfmr1 ( p = 2.067 × 10 –2 ), and between dfmr1 and UH1 -Gal4 driven UAS- 5HT 2A R in dfmr1 ( p = 4.722 × 10 –8 ). For the trained condition, the overall effect of genotype is significant (F(4, 50) = 16.48), with a significant loss between w 1118 and dfmr1 ( p = 1.423 × 10 –2 ), w 1118 and UH1 -Gal4 driven UAS- 5HT 2A R in dfmr1 ( p = 6.309 × 10 –3 ), and between dfmr1 and UH1 -Gal4 driven UAS- 5HT 2A R in the dfmr1 null ( p = 3.127 × 10 –8 ). No significant differences occur in other comparisons. Significance is indicated as p > 0.05 (not significant, ns), p < 0.05 (*), p < 0.01 (**), p < 0.001 (***), and p < 0.0001 (****).

Article Snippet: Primary antibodies used: rabbit anti-5-HT (Immunostar, 20,080 1:1,000), rabbit anti-5HT 2A R (Abcam, ab140524, 1:100), and mouse anti-Trio (Developmental Studies Hybridoma Bank (DSHB), 9.4A, 1:100).

Techniques: Over Expression, Control, Labeling, Fluorescence