Journal: Molecular Brain

Article Title: Exploration of schizophrenia-related behavioral and molecular abnormalities in a mutant mouse model with a mutation in the TVV motif of the ErbB4 gene

doi: 10.1186/s13041-025-01238-2

Figure Lengend Snippet: ErbB4 mutation disrupts key proteins of postsynaptic signaling. ErbB4 is increased in homo mice while phosphorylated ErbB4 is decreased and less activated compared to abundance of total protein amount ( A , B , Student’s t-test, p < 0.0001, p < 0.001, and p < 0.0001 respectively). In contrast, the total amount of PSD-95 is decreased (Student’s t-test, p < 0.05), but the remaining pool is highly activated in mutant mice ( A , B , Student’s t-test, p < 0.05). Mutation also caused hypofunction of NMDAR2A subunit ( A , C ). NMDAR2A is highly decreased (Student’s t-test, p < 0.001) but hyperactivated in mutant homo mice (Student’s t-test, p < 0.01). In contrast, NMDAR1 subunit was not affected which highlights the mutation caused NMDAR subunit specific defects. The GABAergic system is also selectively disrupted ( A , D ). GAD65 a global brain GABAergic maker and VGLUT1 were not affected (Student’s t-test, p > 0.05) but the GAD67 isoform is specifically highly decreased (Student’s t-test, p < 0.01) in mutant homo mice. 60 µg protein, N = 3 and n = 6 in all experiments, * = p < 0.05, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001. * = p < 0.05, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001, N = number of mice, and n = number of samples

Article Snippet: The membranes were blocked with quick blocking buffer (P0252 Beyotime) for 15 min and incubated at 4 °C overnight with the following primary antibodies: rabbit anti-ErbB4 (1:1000, Cat No. 4795, CST), mouse anti-PSD-95 (1:1000, Cat No. MAB1598, Sigma Aldrich), mouse anti-GAPDH (1:5000, Cat No. HC301, TransGen), rabbit anti-phospho-phospho-PSD-95 (1:1000, Cat. No. 45737, CST), rabbit anti-phospho-ErbB4 (1:1000, Cat No. 4757, CST), rabbit anti-NMDA receptor 2 A (1:1000, Cat. No. 4205, CST), rabbit anti-phospho-NMDA receptor 2 A (1:1000, Cat. No. 4206, CST), rabbit anti-GAD67 (1: 2000, Cat. No. ab239372, Abcam), rabbit anti-GAD67 (1: 1000, Cat. No. ab97739, Abcam), mouse anti-NMDAζ1 (1:300, Cat. No. sc-518053, Santa Cruz Biotechnology), and mouse VGLUT1 (1:500, Cat. No. NBP2-59329, Novus Bio).

Techniques: Mutagenesis