Journal: Psychopharmacology

Article Title: Late development of OCD-like phenotypes in Dlgap1 knockout mice

doi: 10.1007/s00213-024-06668-9

Figure Lengend Snippet: Experimental scheme and generation of knockout mice. ( a ) Timeline of observation for skin lesions and behavioral tests. Each triplet of KO, HT and WT littermates was longitudinally observed for skin lesions. At 2 months of age, mice were subjected to modified SHIRPA. If we observed skin lesions with score 2 in the KO mouse (i.e., onset), all members of the triplet were subjected to an open-field test, random assignment to a treatment group, two-week administration of vehicle or fluvoxamine, and 24-hour monitoring of self-grooming behavior, according to the schedule illustrated in the panel. ( b ) Knockout region and primers for genotyping. Red line indicates the knockout region. Forward_2 (F2) and reverse_1 (R1) primers that bind to the regions indicated by the corresponding arrows were used to detect the wild-type allele. Similarly, forward_1 (F1) and R1 primers were used to detect the knockout allele. ( c ) PCR analysis of mouse tail DNA with either of the two pairs of primers (P1: F1/R1 or P2: F2/R1). One of the following three patterns was detected to determine a genotype: WT, detection of a 513 bp band with P2; KO, detection of a 521 bp band with P1; HT, detection of both bands. ( d ) Immunoblots of brain synaptosome fraction probed with anti-DLGAP1 antibody. Specific bands of known sizes (approximately 100 kDa and 130 kDa: Rasmussen et al., 2017) were detected for the samples from WT and HT mice, but not for the sample from a KO mouse. β-tubulin was used as a loading control.

Article Snippet: Membranes were blocked with 5% non-fat dried milk in tris-buffered saline-0.1%/Tween-20 (TBS-0.1/T) for 1 h at room temperature, and then incubated overnight at 4oC with either of the following primary antibodies: anti-DLGAP1 antibody (1:1000 dilution; GeneTex, Irvine, USA Cat#GTX133264) or anti-beta III tubulin antibody (1:1000 dilution; Abcam, Cambridge, UK; Cat# ab18207).

Techniques: Knock-Out, Modification, Western Blot, Control