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. Author manuscript; available in PMC: 2010 Mar 12.
Published in final edited form as: Neuron. 2009 Mar 12;61(5):734–749. doi: 10.1016/j.neuron.2009.01.017

Figure 8. Normal Brain Morphology but Altered VGLUT1 Immunofluorescence in LRRTM1 -/- Mice.

Figure 8

A. Targeting strategy for Lrrtm1 locus. Part of the wild-type Lrrtm1 locus as well as the elements of the replacing cassette are shown. The locations of the restriction enzyme cleavage sites used in the Southern blotting analysis are indicated: B, BamHI; E, EcoRI; IRES, internal ribosome entry site. B. Southern blotting results with a correctly targeted ES cell clone (+/-) and with a wild-type (+/+) ES cell clone are shown. Fragment lengths expected are 6 kb for wild type and 3.6 kb for a correctly targeted allele with the 5′ probe (EcoRI digest), and 10.5 kb for wild type and 5.4 kb for a correctly targeted allele with the 3′ probe (BamHI and SpeI digest). C. Genotyping results for wild-type (+/+) mouse and mice heterozygous (+/-) or homozygous (-/-) for Lrrtm1 deletion. D. No differences were detected in overall cellular or synaptic distribution in the LRRTM1 -/- mouse brain compared with wild type. DAPI stain for nuclei shows normal cellular organization in the hippocampal formation. Immunofluorescence patterns for the presynaptic active zone protein bassoon are indistinguishable between LRRTM -/- and wild type hippocampus. Scale bar = 200 μm.

E-G. Quantitative analysis of immunofluorescence in the hippocampal formation at 4-5 weeks. E. Sample confocal images, here from CA1 stratum radiatum. Scale bar = 10 μm. F. There was no difference in bassoon immunopositive puncta area between LRRTM1 -/- and wild type in any region measured. Measures were made from the same multi-channel images as for panel G. G. Puncta area for the vesicle-associated protein VGLUT1 was selectively increased in CA1 stratum radiatum (Rad) and oriens (Ori) (t-test p<0.01, duplicate experiments of 2 LRRTM1 -/- and 3 wild type mice), but unchanged in LRRTM1 -/- relative to wild type mice in CA1 stratum lacunosum moleculare (SLM) and CA3 stratum lucidum (Luc). Mean values for VGLUT1 puncta area in wild type mice were 1.01 μm2 CA3 Luc, 0.50 μm2 CA1 SLM, 0.49 μm2 CA1 Rad, 0.50 μm2 CA1 Ori. Panels A-C are reproduced from a doctoral dissertation (Lauren, 2007).