Journal: iScience

Article Title: Multimodal single-cell profiling reveals neuronal vulnerability and pathological cell states in focal cortical dysplasia

doi: 10.1016/j.isci.2024.111337

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Article Snippet: Mouse monoclonal anti-MAP2 , Thermo Fisher , Cat#13-1500; RRID: AB_2533001.

Techniques: Recombinant, Gene Expression, Software

Journal: Nature Communications

Article Title: The human VGLUT3-pT8I mutation elicits uneven striatal DA signaling, food or drug maladaptive consumption in male mice

doi: 10.1038/s41467-024-49371-1

Figure Lengend Snippet: a - d , Immunofluorescent detection of VGLUT3 ( a, a’, c, c’, c” ) or VGLUT3-p.T8I ( b, b’, d, d’, d” ) (green) and microtubule-associated protein (MAP2, a, a’, b, b’) or bassoon ( c’, c”, d’, d” ) (red) in hippocampal neuronal culture of WT mice. e , Alignment of human (H) or mouse (M) VGLUT1, VGLUT2 and VGLUT3 amino acid sequences. f , Genotyping of WT (VGLUT3 +/+ ) mice, heterozygous mice (VGLUT3 +/T8I ) or homozygous mice (VGLUT3 T8I/T8I ). g , Schematic representation of the targeting strategy and genotyping strategy of mouse VGLUT3 by PCR. Mice were genotyped with primers (arrows) flanking exon 1 (mf, mr, ex1), in intron 1 (ef) and in the LoxP sites in 3’ of the autoexcision cassette (er). Asterisks represent the targeted site in exon 1. h , Top: Immunoautoradiographic (IAR) regional distribution of VGLUT3 and VGLUT3-p.T8I on coronal sections from the brain of WT mice (black, n = 7 mice) and VGLUT3 T8I/T8I mice (purple, n = 8 mice). Bottom: Densitometric quantification of VGLUT3 and VGLUT3-p.T8I on mouse brain sections. Data are presented as mean values ± SEM. Statistical analysis performed with two-sided unpaired t-test. i-m’ , Immunofluorescent detection of VGLUT3 ( i - m ) or VGLUT3-p.T8I ( i’ - m’ ) in the hippocampus and in the striatum of WT mice or VGLUT3 T8I/T8I mice. n - o , Immunofluorescent detection with STED microscopy of VGLUT3 or VGLUT3-p.T8I (purple) and VAChT (green) in preparations of striatal synaptic vesicles. p , Automatic batch analysis of nearest neighbor distances (NNDs) between VGLUT3- or VGLUT3-p.T8I- and VAChT-immunofluorescent spots on striatal isolated synaptic vesicles. Statistical analysis two-sided Kolmogorov-Smirnov test, p = 0.218. q , Percentage of VGLUT3 or VGLUT3-p.T8I immunofluorescent spots displaying a NND with their closest VAChT-positive spot above and below 95 nm ( p = 0.439, two-sided Chi-squared test). Source data are provided as a Source Data file. Abbreviations: CA1-3, fields of the hippocampus pyramidal layer; Cx, cortex; DG, dentate gyrus; DMS, dorsomedial striatum; DLS, dorsolateral striatum; DR, dorsal raphe nucleus; st, striatum; DSt, dorsal striatum; Hi, hippocampus; IAR, immunoautoradiography; MnR, median raphe nucleus; NAc, nucleus accumbens. Bar: 10 µm in a and b , 5 µm in a’ and b’ , 2 µm in c - d” , 10 µm in i - l’ , 0.1 µm in n and o .

Article Snippet: Coverslips seedings cells or free-floating brain sections were incubated with anti-human VGLUT3 rabbit polyclonal antiserum (1:1000) , anti-VGLUT3 rabbit polyclonal antiserum (1:2000, Synaptic Systems), anti-rodent VGLUT1 rabbit polyclonal antiserum (1:2000) , anti-rodent MAP2 mouse monoclonal antiserum (1:1000, Sigma), anti-rodent bassoon mouse monoclonal antiserum (1:2000, Abcam), or PSD-95 mouse monoclonal antiserum (1:2000, Abcam).

Techniques: Microscopy, Isolation