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. 2016 Dec 13;5:e22757. doi: 10.7554/eLife.22757

Figure 4. Genetic ablation of Nrxn3 AS4 insertion in mice.

(A) Targeting strategy for conditional ablation of Nrxn3 AS4 insertion (Nrxn3ex21ΔPV conditional knock-out). See Experimental Procedures for details. (B) Analysis of Nrxn1,2 and 3 AS4 mRNA in control and in Nrxn3ex21Δ (n = 3 mice for each genotype, mean+SEM, unpaired t-test p<0.0001). (C) Selected Reaction Monitoring MS/MS transitions for an endogenous AS4-specific peptide detected in wild-type and Nrxn3Δex21 knock-out forebrain extracts. Synaptic proteins were enriched in Triton X-100-insoluble fractions from adult mice. The colored lines show signal intensities for several transitions derived from the same peptide. (D) Expression level of Nrx using pan-Nrx antibodies on Triton X-100-insoluble fractions from control (CTRL), Nrxn3ex21Δ knock-out (KO) and Nrxn3ex21Δ/+ heterozygous (HET) mice was determined by Western blotting. (E) Immunolabelling of PV+ interneurons in the CA1 region of hippocampus of control and and Nrxn1/3ex21ΔPV PV conditional double knock-out. PV+ interneurons were identified using anti-PV (magenta in overlay) and Hoechst (green in overlay, to counterstain cell nuclei). Quantifications of PV+ cell density in CA1 per 100 mm2 at postnatal day 24–26. Single dots in the graph represent means of PV+ cells per animal (n = 4 mice for each genotype, mean).

DOI: http://dx.doi.org/10.7554/eLife.22757.010

Figure 4.

Figure 4—figure supplement 1. Analysis of weight and Mendelian frequencies in Nrxn3ex21ΔPV and Nrxn1/3ex21ΔPV conditional knock-out mice.

Figure 4—figure supplement 1.

(A) Weight of control (cre negative) and conditional single (Nrxn3ex21ΔPV, cSKO) and conditional double knock-out mice (Nrxn1/3ex21ΔPV, cDKO) at postnatal day 25. Each dot represents a single animal, mean ± SEM. (B) Single and double conditional knock-out mice are born at Mendelian frequencies (expected frequency was set to 1, for Nrxn three conditional knock-out: 253 mice from 40 independent litters. For Nrxn1/3 conditional knock-out 71 mice from 10 independent litters).
DOI: http://dx.doi.org/10.7554/eLife.22757.011