Journal: Journal of Neuroscience

Article Title: Prostaglandin E2-Induced Masculinization of Brain and Behavior Requires Protein Kinase A, AMPA/Kainate, and Metabotropic Glutamate Receptor Signaling

doi: 10.1523/jneurosci.3603-09.2009

Figure Lengend Snippet: Figure 4. Effect of neonatal administration of inhibitors for the AMPA/kainate and metabotropic glutamate receptors before PGE2 on measures of adult male sexual behavior. The mean SEM of the latency to mount (A), latency to intromission-like behavior (B), number of mounts (C), and number of intromission-like behaviors (D) collapsed across three behavioral trials. Neonatal females coadministered inhibitors of AMPA/kainate and metabotropic glutamate receptors (NBQX and LY341495, re- spectively)beforePGE2didnotexhibitmountingorintromission-likebehaviorsasoftenorinitiatemountingandintromission-like behaviors as quickly as animals treated with VEH PGE2 (*p 0.05 by Tukey’s HSD test), demonstrating a complete block of PGE2-induced masculinization by combined antagonism of AMPA/kainate and metabotropic glutamate receptors. In contrast, masculinized females neonatally treated with only one glutamate receptor inhibitor before PGE2 expressed high levels of sexual behavior in adulthood (*p 0.05 by Tukey’s HSD test, repeated measures ANOVAs; n 5–10).

Article Snippet: Kainate (2.5 g), the AMPA/kainate antagonist 2,3-dihydroxy-6-nitro-7-sulfonyl-benzo[f]quinoxaline (NBQX) (1 g), the type I and II mGluR agonist ( )-1-amino-1,3-cyclopentanedicarboxylic acid (ACPD) (2.28 g), and the type I–III mGluR antagonist LY341495 [(2S)-2-amino-2-[(1S,2S)-2-carboxycycloprop-1-yl]-3-(xanth-9-yl) propanoic acid] (4.65 g) (Tocris Bioscience) were injected subcutaneously in 0.1 ml of 0.9% w/v saline 45 min after the intracerebroventricular injection of 2.5 g of PGE2, 0.9% w/v PBS vehicle, or Ht31.

Techniques: Blocking Assay

Journal: Journal of Neuroscience

Article Title: Prostaglandin E2-Induced Masculinization of Brain and Behavior Requires Protein Kinase A, AMPA/Kainate, and Metabotropic Glutamate Receptor Signaling

doi: 10.1523/jneurosci.3603-09.2009

Figure Lengend Snippet: Figure6. Effectofforskolin,NBQX,andLY341495ontheformationofdendriticspine-likeprocessesonPOAculturedneurons. ForskolinandPGE2treatmentbothinducedathreefoldincreaseinthenumberofdendriticspine-likeprocessesperneurite(A)and per unit length of neurite (B) compared with vehicle treated, whereas cotreatment of NBQX and LY341495 with forskolin pre- ventedtheincreasesuchthatthesemeasureswereequivalenttothoseofvehicle-treatedcells.Photomicrographsofneuritesand dendritic spine-like processes after MAP-2 immunocytochemistry representing the means of the following treatment conditions: PGE2 (C), vehicle (D), forskolin (E), forskolin NBQX LY341495 (F), and NBQX LY341495 (G).

Article Snippet: Kainate (2.5 g), the AMPA/kainate antagonist 2,3-dihydroxy-6-nitro-7-sulfonyl-benzo[f]quinoxaline (NBQX) (1 g), the type I and II mGluR agonist ( )-1-amino-1,3-cyclopentanedicarboxylic acid (ACPD) (2.28 g), and the type I–III mGluR antagonist LY341495 [(2S)-2-amino-2-[(1S,2S)-2-carboxycycloprop-1-yl]-3-(xanth-9-yl) propanoic acid] (4.65 g) (Tocris Bioscience) were injected subcutaneously in 0.1 ml of 0.9% w/v saline 45 min after the intracerebroventricular injection of 2.5 g of PGE2, 0.9% w/v PBS vehicle, or Ht31.

Techniques: Immunocytochemistry

Journal: bioRxiv

Article Title: SRC family kinase inhibition rescues molecular and behavioral phenotypes, but not protein interaction network dynamics, in a mouse model of Fragile X syndrome

doi: 10.1101/2023.06.20.545800

Figure Lengend Snippet: A) Experimental design. B) Principal component graph of median fluorescent intensity (MFI) matrices from N=4 cultures of FMR1 -/y or WT neurons treated with aCSF, NMDA or DHPG. Each colored dot represents an independent agonist-treated cell culture. C) Topological overlap matrix (TOM) plot showing clustering of individual protein interactions into modules. Each point on the dendrograms represents an individual interaction, each pixel of the plot indicates the correlation between the interaction on the X and Y axis (dark = higher correlation coefficient), and the colored boxes outline each module of interest. D) Module-trait correlation table shows the correlation coefficient (top number) and p-value (bottom number) of the correlation between the eigenvector describing each module (indicated by colored bars at left), and the binary-coded hypothesis listed below the table. E) Interactions that were both individually statistically significant by a Bonferroni-corrected adaptive non-parametric test corrected for multiple comparisons comparing treatment groups (ANC, see methods), and were a member of a module of interest, are represented by a row-scaled heatmap (blue-low abundance, orange-high abundance). Interaction labels are colored based on their assigned module. F) Example of an interaction in the NMDA-responsive blue module, Homer1_SAPAP. MFI, median fluorescent intensity. Asterisk represents statistically significant difference measured by ANC. G) Averaged scaled value of all interactions in the NMDA-responsive blue module. Asterisks represent statistically significant difference by one-way ANOVA followed by Sidak’s multiple comparison test, p<0.05. H) Example of an interaction in the DHPG-responsive brown module, CAMKII_Fyn. Asterisk represents statistically significant difference by ANC, p<0.05. I) Averaged scaled value of all interactions in the DHPG-responsive brown module. Asterisks represent statistically significant difference by one-way ANOVA followed by Sidak’s multiple comparison test, p<0.05. J) Example of an interaction in the genotype-correlated yellow module, PI3K_PI3K. Asterisk represents statistically significant difference by ANC, p<0.05. K) Averaged scaled value of all interactions in the genotype-correlated yellow module. Asterisks represent statistically significant difference by one-way ANOVA followed by Sidak’s multiple comparison test, p<0.05. N = 4 cultures from 4 different mice per condition.

Article Snippet: Type I mGluR agonist DHPG (Tocris, 0805; 100 µM) was dissolved in HEPES-aCSF in the presence of non-mGluR glutamate receptor blockers (CNQX, Tocris, 0190, 40 µM; Nimodipine, Tocris, 0600, 1 µM; D(-)-2-Amino-5 phosphonopentanoic acid (APV), Sigma, A5282, 50 µM).

Techniques: Cell Culture, Comparison

Journal: bioRxiv

Article Title: SRC family kinase inhibition rescues molecular and behavioral phenotypes, but not protein interaction network dynamics, in a mouse model of Fragile X syndrome

doi: 10.1101/2023.06.20.545800

Figure Lengend Snippet: A) Experimental design. B-G) Average scaled values of protein interaction modules that respond to NMDA (blue, B-D) and DHPG (brown, E-G) in WT and FMR1 -/y brain slices at P7 (B,E), P17 (C,F) and P60 (D,G). Asterisks represent statistically significant difference by one-way ANOVA followed by Sidak’s multiple comparison test, p<0.05. H-J) Node-edge diagrams show protein interactions that changed in response to DHPG stimulation (compared to aCSF) in WT (H) or FMR1 -/y slices (I) at P17, or interactions that differed comparing FMR1 -/y _aCSF vs. WT_aCSF at p17 (J). Nodes represent proteins, edges represent protein-protein interactions that were significantly different in the given comparison; red = increased, blue = decreased. The relative magnitude of the change is reflected in edge thickness. All interactions shown were both statistically significant by a Bonferroni-corrected adaptive non-parametric test corrected for multiple comparisons (ANC), and were a member of a significant module. N = 4 animals per age/condition.

Article Snippet: Type I mGluR agonist DHPG (Tocris, 0805; 100 µM) was dissolved in HEPES-aCSF in the presence of non-mGluR glutamate receptor blockers (CNQX, Tocris, 0190, 40 µM; Nimodipine, Tocris, 0600, 1 µM; D(-)-2-Amino-5 phosphonopentanoic acid (APV), Sigma, A5282, 50 µM).

Techniques: Comparison

Journal: bioRxiv

Article Title: SRC family kinase inhibition rescues molecular and behavioral phenotypes, but not protein interaction network dynamics, in a mouse model of Fragile X syndrome

doi: 10.1101/2023.06.20.545800

Figure Lengend Snippet: A) Experimental design. B) The averaged scaled value of the NMDA-responsive blue module was significantly reduced following NMDA in all groups. FMR1 -/y showed significant module pre-activation, which was significantly exacerbated by SCB. C) The averaged scaled value of the DHPG-responsive brown module was significantly increased following DHPG in WT slices. In FMR1 -/y slices, the module did not respond to DHPG with or without SCB pretreatment. D) Experimental design. E) Interactions that were both individually statistically significant by a Bonferroni-corrected adaptive non-parametric test corrected for multiple comparisons (ANC) comparing treatment groups, and were a member of a CNA module that correlated with genotype, are represented by a row-scaled heatmap (blue-low abundance, orange-high abundance). Dotted lines divide interactions that were abnormal in FMR1 -/y slices and returned to WT levels by SCB and MPEP (top), those that were changed by SCB and MPEP (middle), and those that remained abnormal despite SCB/MPEP treatment (bottom). F,G) Two interactions were ‘normalized’ by SCB and MPEP treatment, including NMDAR2A_Shank3 (F) and SynGAP_Shank1 (G). MFI, median fluorescent intensity. Asterisks represents statistically significant difference by ANC, p<0.05. N = 4 WT and 4 FMR1 -/y animals; slices from each FMR1 -/y animal were treated with aCSF, SCB and MPEP to directly compare the effects of treatment.

Article Snippet: Type I mGluR agonist DHPG (Tocris, 0805; 100 µM) was dissolved in HEPES-aCSF in the presence of non-mGluR glutamate receptor blockers (CNQX, Tocris, 0190, 40 µM; Nimodipine, Tocris, 0600, 1 µM; D(-)-2-Amino-5 phosphonopentanoic acid (APV), Sigma, A5282, 50 µM).

Techniques: Activation Assay

Journal: Journal of neurochemistry

Article Title: Synaptic activity induces input-specific rearrangements in a targeted synaptic protein interaction network

doi: 10.1111/jnc.14466

Figure Lengend Snippet: (A–D). QMI maps showing network biosignatures for A. 100 μM glutamate, B. 1μM glutamate, C. 100μM NMDA+10μm Glycine, D. 100μm DHPG for N=4–8 biological replicates. (E) Principal component analysis showing separation of NMDA and mGluR treatment along opposing axes, with glutamate intermediate. (F) Topological overlap matrix with significant modules boxed as in Figure 4. (G) Module-trait matrix showing Turquoise module correlated with NMDA stimulation, and the Brown module correlated with DHPG.

Article Snippet: Type I mGluR agonist DHPG (Tocris, 0805; 100μM) was dissolved in HEPES-aCSF in the presence of non-mGluR glutamate receptor blockers (CNQX, Tocris, 0190, 40μM; Nimodipine, Tocris, 0600, 1μM; APV, Sigma, A5282, 50μM).

Techniques: