primary anti pde2a antibody  (Santa Cruz Biotechnology)

Journal: Translational Psychiatry

Article Title: Phosphodiesterase 2A inhibition corrects the aberrant behavioral traits observed in genetic and environmental preclinical models of Autism Spectrum Disorder

doi: 10.1038/s41398-022-01885-2

Figure Lengend Snippet: Fmr1- Δ exon 8 rats emit less USVs when removed from the nest at PNDs 5 ( A ) and 9 ( B ), and this communicative deficit is reversed upon BAY607550 injection (PND 5: WT-VEH, n = 8; WT-BAY, n = 8; KO-VEH, n = 10; KO-BAY, n = 10; PND 9: WT-VEH, n = 8; WT-BAY, n = 8; KO-VEH, n = 9; KO-BAY, n = 9). Fmr1- Δ exon 8 juvenile and adult rats display reduced sociability in the three-chamber test, as they show a lower discrimination index ( C , D ). At both ages, the altered phenotype displayed by Fmr1- Δ exon 8 rats is rescued by PDE2A inhibition (PND 35: WT-VEH, n = 7; WT-BAY, n = 8; KO-VEH, n = 8; KO-BAY, n = 8; PND 90: WT-VEH, n = 11; WT-BAY, n = 8; KO-VEH, n = 11; KO-BAY, n = 12). Data represent mean ± S.E.M. * p < 0.05, ** p < 0.01, *** p < 0.001 vs WT-VEH group, # p < 0.05, ## p < 0.01, ### p < 0.001 vs Fmr1- Δ exon 8 -VEH group (Student’s–Newman–Keuls post hoc test).

Article Snippet: Brain obtained from PND14 VPA and control rats were fixed in 4% ( w / v ) paraformaldehyde for 48 h. Brain slices of 50 μm were obtained by vibratome (Microm HM650V – Thermo Scientific TM, , Illkirch, France) and Immunofluorescence was performed as described [ ], using polyclonal anti-PDE2A primary antibody (# PD2A-101AP, 1:200, FabGennix, Frisco, TX 7) followed by a treatment with 70% True Black (Biotium, USA) diluted in ethanol.

Techniques: Injection, Inhibition

Journal: Translational Psychiatry

Article Title: Phosphodiesterase 2A inhibition corrects the aberrant behavioral traits observed in genetic and environmental preclinical models of Autism Spectrum Disorder

doi: 10.1038/s41398-022-01885-2

Figure Lengend Snippet: A cGMP degradation level was measured in forebrain from VPA and control rats (SAL) at PND 14. Levels of cGMP degradation catalyzed by total extracts in the absence or presence of BAY607550 are shown (Kruskal–Wallis analysis followed by Dunn’s multiple comparisons test; SAL n = 3; SAL + BAY n = 3; VPA n = 3; VPA + BAY n = 3). B PDE2A-specific cGMP degradation measured as the difference of values obtained from forebrain extracts in the absence and presence of BAY607550 (unpaired Student’s t -test, SAL n = 3; VPA n = 3). C At PND 14, mRNA was purified from forebrain of VPA- and SAL-exposed rats. Pde2a mRNA expression level was quantified by RT-qPCR and compared between the two samples (unpaired Student’s t -test, SAL n = 4; VPA n = 4) * p < 0.05; *** p < 0.001. D – F PDE2A expression was quantified by immunofluorescence on brain slices obtained from VPA- and SAL-exposed rats at PND 14. The intensity of immunofluorescence was measured in D cortex, E hippocampal CA3 and F CA1 regions. In upper panels ( D – F ), an example of PDE2A immunofluorescence in the corresponding brain region in SAL- and VPA-treated rats is shown. The intensity of immunofluorescence is indicated as arbitrary unit (A.U.). In lower panels ( D – F ), each graph shows the measure of the intensity of the fluorescence per cell in SAL- and VPA-treated rats. Data represent mean ± S.E.M of the immunofluorescence/cell. Unpaired Student’s t -test. No significant difference was measured. Cortex: 1202 cells analyzed from 8 SAL-treated rat brains and 970 cells analyzed from 8 VPA-treated rat brains; CA3 hippocampal regions: 541 cells analyzed from 8 SAL-treated rat brains and 790 analyzed from 8 VPA-treated rat brains; CA1: 398 cells analyzed from 7 SAL-treated rat brains and 899 cell analyzed from 10 VPA-treated rat brains.

Article Snippet: Brain obtained from PND14 VPA and control rats were fixed in 4% ( w / v ) paraformaldehyde for 48 h. Brain slices of 50 μm were obtained by vibratome (Microm HM650V – Thermo Scientific TM, , Illkirch, France) and Immunofluorescence was performed as described [ ], using polyclonal anti-PDE2A primary antibody (# PD2A-101AP, 1:200, FabGennix, Frisco, TX 7) followed by a treatment with 70% True Black (Biotium, USA) diluted in ethanol.

Techniques: Control, Purification, Expressing, Quantitative RT-PCR, Immunofluorescence, Fluorescence

Journal: Translational Psychiatry

Article Title: Phosphodiesterase 2A inhibition corrects the aberrant behavioral traits observed in genetic and environmental preclinical models of Autism Spectrum Disorder

doi: 10.1038/s41398-022-01885-2

Figure Lengend Snippet: PDE2A inhibition by BAY607550 normalizes the altered USV profile displayed by VPA-exposed pups both PNDs 5 ( A ) and 9 ( B ) (PND 5: SAL-VEH, n = 7; SAL-BAY, n = 6; VPA-VEH, n = 10; VPA-BAY, n = 10, PND 9: SAL-VEH, n = 7; SAL-BAY, n = 6; VPA-VEH, n = 10; VPA-BAY, n = 10). Moreover, administration of BAY607550 rescues the lower discrimination index displayed by VPA-exposed rats in the three-chamber test ( C ) (SAL-VEH, n = 8; SAL-BAY, n = 5; VPA-VEH, n = 7; VPA-BAY, n = 8). At adulthood, VPA-exposed rats present impaired consolidation of aversive memories, that is reversed following administration of BAY607550 ( D ) (SAL-VEH, n = 31; SAL-BAY, n = 30; VPA-VEH, n = 16; VPA-BAY, n = 16). Data represent mean ± S.E.M. * p < 0.05 vs SAL-VEH group, # p < 0.05, ## p < 0.01 vs Fmr1- Δ exon 8 -VEH group (Student’s–Newman–Keuls post hoc test); ** p < 0.01, *** p < 0.001 vs acquisition time (Student’s–Newman–Keuls post hoc test).

Article Snippet: Brain obtained from PND14 VPA and control rats were fixed in 4% ( w / v ) paraformaldehyde for 48 h. Brain slices of 50 μm were obtained by vibratome (Microm HM650V – Thermo Scientific TM, , Illkirch, France) and Immunofluorescence was performed as described [ ], using polyclonal anti-PDE2A primary antibody (# PD2A-101AP, 1:200, FabGennix, Frisco, TX 7) followed by a treatment with 70% True Black (Biotium, USA) diluted in ethanol.

Techniques: Inhibition

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Phosphodiesterase 2 inhibition preferentially promotes NO/guanylyl cyclase/cGMP signaling to reverse the development of heart failure.

doi: 10.1073/pnas.1800996115

Figure Lengend Snippet: Fig. 8. PDE2 inhibition augments cardiac cGMP, but not cAMP, levels in vivo and promotes the antihypertrophic effect of NO, but not ANP, in isolated cardiomyocytes. (A) Cardiomyocyte area in cells isolated from neonatal WT mice treated with Ang II (100 nM) for 24 h in the absence and presence of the NO donor DETA (10 μM), ANP (1 μM), or BNP (1 μM) under basal conditions or following treatment with BAY 60-7550 (100 nM). (B) Cyclic nucleotide levels in whole-heart homogenates from sham mice or animals subjected to AAC for 6 wk (6w) in the absence and presence of BAY 60-7550 (10 mg·kg−1·d−1 p.o., initiated at 3 wk). (C) Immunoblot analysis of PDE2A expression in whole-heart homogenates from mice subjected to AAC for 6w in the absence and presence of BAY 60-7550 (10 mg·kg−1·d−1 p.o., initiated at 3 wk). Data are expressed as mean ± SEM with analysis by one-way ANOVA with a Bonferroni post hoc test. *P < 0.05 versus Ang II + DETA (A); **P < 0.01 versus sham versus ACC, #P < 0.05 versus AAC (B); *P < 0.05 versus sham (C) (n = 6).

Article Snippet: PDE2A protein expression was determined by immunoblot using primary anti-PDE2A antibody (1:500; Santa Cruz Biotechnology) and secondary horseradish peroxidase-conjugated anti-goat IgG antibody (1:10,000; Santa Cruz Biotechnology).

Techniques: Inhibition, In Vivo, Isolation, Western Blot, Expressing