Fig. 5: Generation of Slitrk2-cKO mice. | Nature Communications

Fig. 5: Generation of Slitrk2-cKO mice.

From: SLITRK2 variants associated with neurodevelopmental disorders impair excitatory synaptic function and cognition in mice

Fig. 5: Generation of Slitrk2-cKO mice.The alternative text for this image may have been generated using AI.

a Strategy used to generate Slitrk2-cKO mice. LoxP sites were introduced at positions flanking the neomycin gene, FLP recombinant target (FRT), and exon 2–3 (E2 and E3) of the Slitrk2 gene. Black arrows indicate forward and reverse primers used for genotyping. Note that lacZ and neomycin cassettes are two separate markers. b PCR genotyping of Slitrk2-floxed mice (top) and Nestin-Cre;Slitrk2-cKO (Nestin-Slitrk2) mice (bottom). The band size for the Slitrk2-floxed allele was 245 bp. The 400-bp Cre-specific PCR product was detected only in Nestin-Slitrk2 mice. c Quantitative RT-PCR analyses of Slitrk mRNA levels in Nestin-Slitrk2 mice. Data are mean ± SEMs (n = 4 mice; *p < 0.05; two-tailed Mann–Whitney U test). d Body weights of Nestin-Slitrk2 and Slitrk2-floxed (Ctrl) mice at P30, P40, and P50. Data are means ± SEMs (‘n’ denotes the number of mice; P30: Ctrl, n = 15; Nestin-Slitrk2, n = 10; P40: Ctrl, n = 20; Nestin-Slitrk2, n = 14; P50: Ctrl, n = 20; Nestin-Slitrk2, n = 14; *p < 0.05, ****p < 0.0001; two-tailed Mann–Whitney U test). e, f Representative photographs of whole brains (e) and quantification of brain weights and size (f) of P42 Nestin-Slitrk2 and Slitrk2-floxed (Ctrl) mice. Data are means ± SEMs (‘n’ denotes the number of mice; Ctrl and Nestin-Slitrk2, n = 5; two-tailed Mann–Whitney U test). g Normal gross morphology of the Nestin-Slitrk2 brain at 7 wks, as revealed by Nissl staining. Scale bar: 1 mm (applies to both top and bottom). h Representative images of NeuN (a neuronal marker) staining in the Nestin-Slitrk2 brain. Normal numbers of neurons in hippocampal regions. Top, hippocampus; bottom, CA1 stratum pyramidale layer. Scale bar: 1 mm (top) and 20 µm (bottom). i Summary graphs for neuron density and cortical thickness. Data are means ± SEMs (‘n’ denotes the number of mice; neuron density: Ctrl and Nestin-Slitrk2, n = 4; cortical thickness: Ctrl and Nestin-Slitrk2, n = 6; two-tailed Mann–Whitney U test). j Representative images of GAD67 (GABAergic interneuron marker) staining in the hippocampal CA1 of Nestin-Slitrk2 mice. Scale bar: 0.1 mm. k Summary data for GAD67 staining from (j). Data are means ± SEMs (n = 3 mice each after averaging data from 3 sections/mouse; two-tailed Mann–Whitney U test). l, m Representative immunoblots (l) of hippocampal lysates, and summary data showing synaptic protein levels (m) in crude synaptosomal fractions of P42 control and Nestin-Slitrk2 brains, analyzed by semi-quantitative immunoblotting. Data are means ± SEMs (n = 4 mice/group; *p < 0.05, ***p < 0.001; two-tailed unpaired t-test). See Source data for raw data values and Supplementary Table 4 for statistical details.

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