Journal: Molecular Psychiatry

Article Title: ZFP804A mutant mice display sex-dependent schizophrenia-like behaviors

doi: 10.1038/s41380-020-00972-4

Figure Lengend Snippet: A PPI values for wild-type control and ZFP804A mutant mice at three different prepulse intensities (65, 73, and 82 dB) at the age of 8 months. Significantly reduced PPI is observed in female ZFP804A mutant mice compared with female controls ( P 65dB = 0.0044**; P 73dB = 0.0088**; P 82dB = 0.0004***), and with male ZFP804A mutant mice ( P 73dB = 0.0397*; P 82dB = 0.0205*). n = 9–10 in each group. Two-way ANOVA with Bonferroni correction analysis. At 65 dB, genotyping effect: F [1, 33] = 11.35, P = 0.0019**; sex effect: F [1, 33] = 4.170, P = 0.0492*; interaction: F [1, 33] = 3.848, P = 0.0583. At 73 dB, genotyping effect: F [1, 33] = 8.927, P = 0.0053**; gender factor: F [1, 33] = 4.706, P = 0.0374*; interaction: F [1, 33] = 3.928, P = 0.0559. At 82 dB, genotyping effect: F [1, 33] = 17.71, P = 0.0001***; sex effect: F [1, 33] = 4.658, P = 0.0383*; interaction: F [1, 33] = 5.574, P = 0.0243*. B Impaired PPI is restored in female ZFP804A mutant mice by the administration of clozapine ( P 73dB = 0.0025**; P 82dB = 0.0004***). n = 9–10 in each group. One-way ANOVA with Bonferroni correction analysis. At 65 dB, treatment effect: F [3, 35] = 3.168, P = 0.0363*. At 73 dB, treatment effect: F [3, 35] = 7.087, P = 0.0008***. At 82 dB, treatment effect: F [3, 35] = 7.721, P = 0.0004***. C PPI values are normal in ovariectomized female ZFP804A mutant mice compared with female control mice at three different prepulse intensities (65, 73, and 82 dB) at the age of 8 months. n = 8–9 in each group. Student’s t test analysis ( t 65dB = 0.1673, P 65dB = 0.8693; t 73dB = 0.9923, P 73dB = 0.3368; t 82dB = 0.8783, P 82dB = 0.3936). D Representative blots for protein detected by indicated antibodies in the hippocampal tissues from control and ZFP804A mutant mice at the age of 6 months. E Quantification of relative levels of proteins as normalized to GAPDH or β-Tubulin protein expression from male hippocampus at the age of 6 months. n = 3 per protein per genotype. One-sample t- test ( t D1R = 1.757, P D1R = 0.153; t D2R = 0.265, P D2R = 0.803; t D5R = 0.175, P D5R = 0.868; t 5HT1bR = 1.149, P 5HT1bR = 0.314; t 5HT2aR = 0.015, P 5HT2aR = 0.988; t 5HT2cR = 0.867, P 5HT2cR = 0.434; t 5HT3aR = 0.577, P 5HT3aR = 0.594; t 5HT6R = 0.119, P 5HT6R = 0.91; t 5HT7R = 0.061, P 5HT7R = 0.954; t GABAB1R = 0.481; P GABAB1R = 0.655; t GABAB2R = 0.213, P GABAB2R = 0.841). F Quantification of relative levels of proteins as normalized to GAPDH or β-Tubulin protein expression from female hippocampus at the age of 6 months. n = 3 per protein per genotype. One-sample t- test ( t D1R = 0.831, P D1R = 0.452; t D2R = 0.808, P D2R = 0.464; t D5R = 0.05, P D5R = 0.962; t 5HT1bR = 1.113, P 5HT1bR = 0.328; t 5HT2aR = 0.721, P 5HT2aR = 0.510; t 5HT2cR = 0.272, P 5HT2cR = 0.799; t 5HT3aR = 0.913, P 5HT3aR = 0.412; t 5HT4R = 0.201, P 5HT6R = 0.014*; t 5HT7R = 0.102, P 5HT7R = 0.923; t GABAB1R = 6.138, P GABAB1R = 0.003**; t GABAB2R = 0.4129, P GABAB2R = 0.700). G Representative blots for protein detected by indicated antibodies in the cortical tissues from control and ZFP804A mutant mice at the age of 6 months. H Quantification of relative levels of proteins as normalized to GAPDH or β-Tubulin protein expression from male cortex at the age of 6 months. n = 3 per protein per genotype. One-sample t- test ( t D1R = 0.128, P D1R = 0.903; t D2R = 1.306, P D2R = 0.261; t D5R = 0.236, P D5R = 0.824; t 5HT1bR = 0.785, P 5HT1bR = 0.476; t 5HT2aR = 0.175, P 5HT2aR = 0.869; t 5HT2cR = 0.317, P 5HT2cR = 0.767; t 5HT3aR = 2.117, P 5HT3aR = 0.101; t 5HT6R = 1.369, P 5HT6R = 0.242; t 5HT7R = 1.280, P 5HT7R = 0.269; P GABAB1R = 0.650; t GABAB2R = 1.813, P GABAB2R = 0.144). I Quantification of relative levels of proteins as normalized to GAPDH or β-Tubulin protein expression from female cortex at the age of 6 months. n = 3 per protein per genotype. One-sample t- test ( t D1R = 1.484, P D1R = 0.211; t D2R = 6.086, P D2R = 0.003**; t D5R = 1.131, P D5R = 0.321; t 5HT1bR = 1.473, P 5HT1bR = 0.214; t 5HT2aR = 0.231, P 5HT2aR = 0.828; t 5HT2cR = 0.061, P 5HT2cR = 0.954; t 5HT3aR = 0.834, P 5HT3aR = 0.451; t 5HT6R = 0.277, P 5HT6R = 0.794; t 5HT7R = 0.062, P 5HT7R = 0.952; t GABAB1R = 0.748, P GABAB1R = 0.495; t GABAB2R = 1.160, P GABAB2R = 0.310). J None hippocampal CA1 LTD is induced by low-frequency stimulation (LFS) in male control and male ZFP804A mutant mice. n = 7 cells from 3 mice at the age of 2 months in each group. Student’s t test ( t = 0.3271, P = 0.7481). K Hippocampal CA1 LTD is induced by LFS in female ZFP804A mutant mice, but no in female control mice. n = 8–9 cells from 3 mice at the age of 2 months in each group. Student’s t test ( t = 4.245, P = 0.0007***). In addition, a significant difference of hippocampal CA1 LTD induced by LFS is also observed between control and ZFP804A mutant mice with the genders via two-way ANOVA with Bonferroni correction analysis. Genotyping effect: F [1, 30] = 5.081, P = 0.0316*; gender factor: F [1, 30] = 0.0909, P = 0.7651; interaction: F [1, 30] = 7.733, P = 0.0093**. M male mice, F female mice, Con wild-type controls, Mu ZFP804A mutant mice.

Article Snippet: The following primary antibodies were used: rat anti-D1R (1:500, #D2944, Sigma), rabbit anti-D2R (1:300, #55084-1-AP, Proteintech), rabbit anti-D5R (1:1000, #ab181623, Abcam), rabbit anti-5-HT1bR (1:100, #ab13869, Abcam), rabbit anti-5-HT2aR (1:100, #24288, Immuno Star), rabbit anti-5-HT2cR (1:100, #24505, Immuno Star), rabbit anti-5-HT3aR (1:500, #NB100-56351, NOVUS), rabbit anti-5-HT6R (1:500, #24507, Immuno Star), rabbit anti-5-HT7R (1:200, #13830-1-AP, Proteintech), rabbit anti-GABAB1R (1:1000, #ab16604, Abcam), rabbit anti-GABAB2R (1:1000, #ab75838, Abcam), and mouse anti-GAPDH (1:2000, #LF206, EpiZyme) or rabbit anti-β-Tubulin (1:1000, #LF202, EpiZyme) was used as an internal control.

Techniques: Mutagenesis, Expressing

Journal: Molecular Psychiatry

Article Title: ZFP804A mutant mice display sex-dependent schizophrenia-like behaviors

doi: 10.1038/s41380-020-00972-4

Figure Lengend Snippet: A PPI values for wild-type control and ZFP804A mutant mice at three different prepulse intensities (65, 73, and 82 dB) at the age of 8 months. Significantly reduced PPI is observed in female ZFP804A mutant mice compared with female controls ( P 65dB = 0.0044**; P 73dB = 0.0088**; P 82dB = 0.0004***), and with male ZFP804A mutant mice ( P 73dB = 0.0397*; P 82dB = 0.0205*). n = 9–10 in each group. Two-way ANOVA with Bonferroni correction analysis. At 65 dB, genotyping effect: F [1, 33] = 11.35, P = 0.0019**; sex effect: F [1, 33] = 4.170, P = 0.0492*; interaction: F [1, 33] = 3.848, P = 0.0583. At 73 dB, genotyping effect: F [1, 33] = 8.927, P = 0.0053**; gender factor: F [1, 33] = 4.706, P = 0.0374*; interaction: F [1, 33] = 3.928, P = 0.0559. At 82 dB, genotyping effect: F [1, 33] = 17.71, P = 0.0001***; sex effect: F [1, 33] = 4.658, P = 0.0383*; interaction: F [1, 33] = 5.574, P = 0.0243*. B Impaired PPI is restored in female ZFP804A mutant mice by the administration of clozapine ( P 73dB = 0.0025**; P 82dB = 0.0004***). n = 9–10 in each group. One-way ANOVA with Bonferroni correction analysis. At 65 dB, treatment effect: F [3, 35] = 3.168, P = 0.0363*. At 73 dB, treatment effect: F [3, 35] = 7.087, P = 0.0008***. At 82 dB, treatment effect: F [3, 35] = 7.721, P = 0.0004***. C PPI values are normal in ovariectomized female ZFP804A mutant mice compared with female control mice at three different prepulse intensities (65, 73, and 82 dB) at the age of 8 months. n = 8–9 in each group. Student’s t test analysis ( t 65dB = 0.1673, P 65dB = 0.8693; t 73dB = 0.9923, P 73dB = 0.3368; t 82dB = 0.8783, P 82dB = 0.3936). D Representative blots for protein detected by indicated antibodies in the hippocampal tissues from control and ZFP804A mutant mice at the age of 6 months. E Quantification of relative levels of proteins as normalized to GAPDH or β-Tubulin protein expression from male hippocampus at the age of 6 months. n = 3 per protein per genotype. One-sample t- test ( t D1R = 1.757, P D1R = 0.153; t D2R = 0.265, P D2R = 0.803; t D5R = 0.175, P D5R = 0.868; t 5HT1bR = 1.149, P 5HT1bR = 0.314; t 5HT2aR = 0.015, P 5HT2aR = 0.988; t 5HT2cR = 0.867, P 5HT2cR = 0.434; t 5HT3aR = 0.577, P 5HT3aR = 0.594; t 5HT6R = 0.119, P 5HT6R = 0.91; t 5HT7R = 0.061, P 5HT7R = 0.954; t GABAB1R = 0.481; P GABAB1R = 0.655; t GABAB2R = 0.213, P GABAB2R = 0.841). F Quantification of relative levels of proteins as normalized to GAPDH or β-Tubulin protein expression from female hippocampus at the age of 6 months. n = 3 per protein per genotype. One-sample t- test ( t D1R = 0.831, P D1R = 0.452; t D2R = 0.808, P D2R = 0.464; t D5R = 0.05, P D5R = 0.962; t 5HT1bR = 1.113, P 5HT1bR = 0.328; t 5HT2aR = 0.721, P 5HT2aR = 0.510; t 5HT2cR = 0.272, P 5HT2cR = 0.799; t 5HT3aR = 0.913, P 5HT3aR = 0.412; t 5HT4R = 0.201, P 5HT6R = 0.014*; t 5HT7R = 0.102, P 5HT7R = 0.923; t GABAB1R = 6.138, P GABAB1R = 0.003**; t GABAB2R = 0.4129, P GABAB2R = 0.700). G Representative blots for protein detected by indicated antibodies in the cortical tissues from control and ZFP804A mutant mice at the age of 6 months. H Quantification of relative levels of proteins as normalized to GAPDH or β-Tubulin protein expression from male cortex at the age of 6 months. n = 3 per protein per genotype. One-sample t- test ( t D1R = 0.128, P D1R = 0.903; t D2R = 1.306, P D2R = 0.261; t D5R = 0.236, P D5R = 0.824; t 5HT1bR = 0.785, P 5HT1bR = 0.476; t 5HT2aR = 0.175, P 5HT2aR = 0.869; t 5HT2cR = 0.317, P 5HT2cR = 0.767; t 5HT3aR = 2.117, P 5HT3aR = 0.101; t 5HT6R = 1.369, P 5HT6R = 0.242; t 5HT7R = 1.280, P 5HT7R = 0.269; P GABAB1R = 0.650; t GABAB2R = 1.813, P GABAB2R = 0.144). I Quantification of relative levels of proteins as normalized to GAPDH or β-Tubulin protein expression from female cortex at the age of 6 months. n = 3 per protein per genotype. One-sample t- test ( t D1R = 1.484, P D1R = 0.211; t D2R = 6.086, P D2R = 0.003**; t D5R = 1.131, P D5R = 0.321; t 5HT1bR = 1.473, P 5HT1bR = 0.214; t 5HT2aR = 0.231, P 5HT2aR = 0.828; t 5HT2cR = 0.061, P 5HT2cR = 0.954; t 5HT3aR = 0.834, P 5HT3aR = 0.451; t 5HT6R = 0.277, P 5HT6R = 0.794; t 5HT7R = 0.062, P 5HT7R = 0.952; t GABAB1R = 0.748, P GABAB1R = 0.495; t GABAB2R = 1.160, P GABAB2R = 0.310). J None hippocampal CA1 LTD is induced by low-frequency stimulation (LFS) in male control and male ZFP804A mutant mice. n = 7 cells from 3 mice at the age of 2 months in each group. Student’s t test ( t = 0.3271, P = 0.7481). K Hippocampal CA1 LTD is induced by LFS in female ZFP804A mutant mice, but no in female control mice. n = 8–9 cells from 3 mice at the age of 2 months in each group. Student’s t test ( t = 4.245, P = 0.0007***). In addition, a significant difference of hippocampal CA1 LTD induced by LFS is also observed between control and ZFP804A mutant mice with the genders via two-way ANOVA with Bonferroni correction analysis. Genotyping effect: F [1, 30] = 5.081, P = 0.0316*; gender factor: F [1, 30] = 0.0909, P = 0.7651; interaction: F [1, 30] = 7.733, P = 0.0093**. M male mice, F female mice, Con wild-type controls, Mu ZFP804A mutant mice.

Article Snippet: The following primary antibodies were used: rat anti-D1R (1:500, #D2944, Sigma), rabbit anti-D2R (1:300, #55084-1-AP, Proteintech), rabbit anti-D5R (1:1000, #ab181623, Abcam), rabbit anti-5-HT1bR (1:100, #ab13869, Abcam), rabbit anti-5-HT2aR (1:100, #24288, Immuno Star), rabbit anti-5-HT2cR (1:100, #24505, Immuno Star), rabbit anti-5-HT3aR (1:500, #NB100-56351, NOVUS), rabbit anti-5-HT6R (1:500, #24507, Immuno Star), rabbit anti-5-HT7R (1:200, #13830-1-AP, Proteintech), rabbit anti-GABAB1R (1:1000, #ab16604, Abcam), rabbit anti-GABAB2R (1:1000, #ab75838, Abcam), and mouse anti-GAPDH (1:2000, #LF206, EpiZyme) or rabbit anti-β-Tubulin (1:1000, #LF202, EpiZyme) was used as an internal control.

Techniques: Mutagenesis, Expressing