gaba a receptor antagonist dmcm  (Tocris)

Journal: Frontiers in Cellular Neuroscience

Article Title: GABA B Receptor-Mediated Impairment of Intermediate Progenitor Maturation During Postnatal Hippocampal Neurogenesis of Newborn Rats

doi: 10.3389/fncel.2021.651072

Figure Lengend Snippet: Representative hippocampal paraffin sections (A) of control animals, DMCM hydrochloride (DMCM) in doses of either 2 μg/kg, 10 μg/kg or 50 μg/kg, and CGP 35348 in doses of either 0.4 mg/kg, 2 mg/kg or 10 mg/kg treated rat pups at P11 co-labeled with DAPI, Nestin, and PCNA. Application of GABA B receptor antagonist CGP 2 mg/kg decreased Nestin positive progenitor cells in the dentate gyrus (DG). Application of CGP 10 mg/kg decreased the number of proliferating Nestin/PCNA double positive cells. Quantification of (B) Nestin and Nestin/PCNA double positive cells in sum of the DG in comparison to control group (100% white bars). Data are expressed relative to the control group as mean ± SEM of n = 10 each group. The 100% values are for Nestin+ 83.5 cell counts and for Nestin+PCNA+ 13.3 cell counts. * p < 0.05 and ** p < 0.01 vs. control (Brown-Forsythe test for Nestin+, Kruskal–Wallis test for Nestin+PCNA+). Expressions of (C) glial fibrillary acidic protein (GFAP) , Scl1a3 , Hes5 , and Sox2 are not affected by the application of DMCM or CGP. Pax6 expression is diminished in CGP treated animals. The relative mRNA expressions of markers were measured by quantitative real-time PCR in rat brain homogenates with DMCM 50 μg/kg (gray bars) or CGP 10 mg/kg (black bars) application relative to control (white bars). Bars represent the relative mRNA quantification based on internal standard HPRT . Data shown as mean ± SEM, n = 9–10. ** p < 0.01 vs. control (Brown-Forsythe test for GFAP , one-way analysis of variance (ANOVA) for Scl1a3 , Hes5 , Sox2 , and Pax6 ).

Article Snippet: Rat pups were cross-gender randomly assigned into a control group with 0.9% saline and six verum groups with GABA A receptor antagonist DMCM (4-Ethyl-6,7-dimethoxy-9H-pyrido[3,4-b]indole-3-carboxylic acid methyl ester) hydrochloride administered at three dosages (2 μg/kg, 10 μg/kg, or 50 μg/kg body weight; Tocris, cat. no. 3083, Wiesbaden-Nordenstadt, Germany), and with GABA B receptor antagonist CPG 35348 at 0.4 mg/kg, 2 mg/kg, or 10 mg/kg body weight (Tocris, cat. no. 1245), respectively.

Techniques: Control, Labeling, Comparison, Expressing, Real-time Polymerase Chain Reaction

Journal: Frontiers in Cellular Neuroscience

Article Title: GABA B Receptor-Mediated Impairment of Intermediate Progenitor Maturation During Postnatal Hippocampal Neurogenesis of Newborn Rats

doi: 10.3389/fncel.2021.651072

Figure Lengend Snippet: Representative hippocampal paraffin sections (A) of control animals, DMCM hydrochloride (DMCM) in doses of either 2 μg/kg, 10 μg/kg or 50 μg/kg, and CGP 35348 in doses of either 0.4 mg/kg, 2 mg/kg or 10 mg/kg treated rat pups at P11 co-labeled with DAPI, NeuroD1, and PCNA. Application of GABA B receptor antagonist CGP decreased NeuroD1 positive progenitor cells in the DG and NeuroD1/PCNA double positive cells in the group with the highest dose of 10 mg/kg. Quantification of (B) NeuroD1 and NeuroD1/PCNA double positive cells in sum of the DG in comparison to control group (100% white bars). Data are expressed relative to the control group as mean ± SEM of n = 10 each group. The 100% values are for NeuroD1+ 308.9 cell counts and for NeuroD1+PCNA+ 7.7 cell counts. * p < 0.05 and **** p < 0.0001 vs. control (one-way ANOVA for NeuroD1+, Brown-Forsythe test for NeuroD1+PCNA+). Expressions of (C) Ascl1 and NeuroD1 are reduced in CGP treated animals and expression of Tbr2 is increased in DMCM treated animals. Ngn2 does not get affected by GABA receptor antagonists. The relative mRNA expressions of markers were measured by quantitative real-time PCR in rat brain homogenates with DMCM 50 μg/kg (gray bars) or CGP 10 mg/kg (black bars) application relative to control (white bars). Bars represent the relative mRNA quantification based on internal standard HPRT . Data shown as mean ± SEM, n = 9–10. * p < 0.05 and ** p < 0.01 vs. control (one-way ANOVA for Ascl1 and Tbr2 , Kruskal–Wallis test for NeuroD1 , Brown-Forsythe test for Ngn2 ).

Article Snippet: Rat pups were cross-gender randomly assigned into a control group with 0.9% saline and six verum groups with GABA A receptor antagonist DMCM (4-Ethyl-6,7-dimethoxy-9H-pyrido[3,4-b]indole-3-carboxylic acid methyl ester) hydrochloride administered at three dosages (2 μg/kg, 10 μg/kg, or 50 μg/kg body weight; Tocris, cat. no. 3083, Wiesbaden-Nordenstadt, Germany), and with GABA B receptor antagonist CPG 35348 at 0.4 mg/kg, 2 mg/kg, or 10 mg/kg body weight (Tocris, cat. no. 1245), respectively.

Techniques: Control, Labeling, Comparison, Expressing, Real-time Polymerase Chain Reaction

Journal: Frontiers in Cellular Neuroscience

Article Title: GABA B Receptor-Mediated Impairment of Intermediate Progenitor Maturation During Postnatal Hippocampal Neurogenesis of Newborn Rats

doi: 10.3389/fncel.2021.651072

Figure Lengend Snippet: Representative hippocampal paraffin sections (A) of control animals, DMCM hydrochloride (DMCM) in doses of either 2 μg/kg, 10 μg/kg or 50 μg/kg, and CGP 35348 in doses of either 0.4 mg/kg, 2 mg/kg or 10 mg/kg treated rat pups at P11 co-labeled with DAPI, NeuN, and PCNA. Application of GABA receptor antagonists did not affect cell counts for postmitotic NeuN+ neurons at the DG. Application of CGP 2 mg/kg led to an increased number of proliferating PCNA+ cells. Quantification of (B) NeuN and PCNA positive cells in sum of the DG in comparison to control group (100% white bars). Data are expressed relative to the control group as mean ± SEM of n = 10 each group. The 100% values are for NeuN+ 143.0 cell counts and for PCNA+ 81.8 cell counts. * p < 0.05 vs. control (Brown-Forsythe test). Expressions of (C) Tbr1 and NeuroD2 are reduced and expression of CycD2 is increased in CGP treated animals. Expression of Tbr1 is increased in DMCM treated animals. GABA receptor antagonists do not affect the expression of Prox1 . The relative mRNA expressions of markers were measured by quantitative real-time PCR in rat brain homogenates with DMCM 50 μg/kg (gray bars) or CGP 10 mg/kg (black bars) application relative to control (white bars). Bars represent the relative mRNA quantification based on internal standard HPRT . Data shown as mean ± SEM, n = 9–10. ** p < 0.01 and *** p < 0.001 vs. control (Brown-Forsythe test for CycD2 and NeuroD2 , one-way ANOVA for Tbr1 ).

Article Snippet: Rat pups were cross-gender randomly assigned into a control group with 0.9% saline and six verum groups with GABA A receptor antagonist DMCM (4-Ethyl-6,7-dimethoxy-9H-pyrido[3,4-b]indole-3-carboxylic acid methyl ester) hydrochloride administered at three dosages (2 μg/kg, 10 μg/kg, or 50 μg/kg body weight; Tocris, cat. no. 3083, Wiesbaden-Nordenstadt, Germany), and with GABA B receptor antagonist CPG 35348 at 0.4 mg/kg, 2 mg/kg, or 10 mg/kg body weight (Tocris, cat. no. 1245), respectively.

Techniques: Control, Labeling, Comparison, Expressing, Real-time Polymerase Chain Reaction

Journal: Frontiers in Cellular Neuroscience

Article Title: GABA B Receptor-Mediated Impairment of Intermediate Progenitor Maturation During Postnatal Hippocampal Neurogenesis of Newborn Rats

doi: 10.3389/fncel.2021.651072

Figure Lengend Snippet: Expression of neurotrophins BDNF , NGF , and NT-3 is reduced in CGP treated animals. The relative mRNA expression of markers was measured by quantitative real-time PCR in rat brain homogenates with DMCM 50 μg/kg (gray bars) or CGP 10 mg/kg (black bars) application relative to control (white bars). Bars represent the relative mRNA quantification based on internal standard HPRT . Data shown as mean ± SEM, n = 9–10. * p < 0.05, ** p < 0.01, and *** p < 0.001 vs. control (Brown-Forsythe test for BDNF , one-way ANOVA for NGF and NT-3 ).

Article Snippet: Rat pups were cross-gender randomly assigned into a control group with 0.9% saline and six verum groups with GABA A receptor antagonist DMCM (4-Ethyl-6,7-dimethoxy-9H-pyrido[3,4-b]indole-3-carboxylic acid methyl ester) hydrochloride administered at three dosages (2 μg/kg, 10 μg/kg, or 50 μg/kg body weight; Tocris, cat. no. 3083, Wiesbaden-Nordenstadt, Germany), and with GABA B receptor antagonist CPG 35348 at 0.4 mg/kg, 2 mg/kg, or 10 mg/kg body weight (Tocris, cat. no. 1245), respectively.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Control