Distinct but overlapping roles of LRRTM1 and LRRTM2 in developing and mature hippocampal circuits
- Neuroscience Research Program, Kleysen Institute for Advanced Medicine, Health Sciences Centre, Canada
- Department of Physiology and Pathophysiology, University of Manitoba, Canada
- Department of Psychiatry and Djavad Mowafaghian Centre for Brain Health, University of British Columbia, Canada
- Department of Biology, York University, Canada
- Department of Cellular and Physiological Sciences, University of British Columbia, Canada
- Department of Molecular Neurobiology, Max Planck Institute for Multidisciplinary Sciences, Germany
- Division of Pathogenetic Signaling, Department of Biochemistry and Molecular Biology, Kobe University Graduate School of Medicine, Japan
- Department of Gerontology, Laboratory of Molecular Life Science, Institute of Biomedical Research and Innovation, Foundation for Biomedical Research and Innovation at Kobe, Japan
- Department of Pharmacology, Gunma University Graduate School of Medicine, Japan
- Division of Neurology, Department of Medicine and Djavad Mowafaghian Centre for Brain Health, University of British Columbia, Canada
- Department of Pharmacology and Therapeutics, University of Manitoba, Canada
- The Children's Hospital Research Institute of Manitoba, Canada
- Program in Biomedical Engineering, University of Manitoba, Canada
Figures
Figure 1 with 3 supplements
LRRTM1 and LRRTM2 control excitatory presynapse development in the hippocampus.
(A) Confocal immunofluorescence images for VGlut1, GAD65 and the nuclear marker DAPI revealed normal hippocampal morphology and large-scale synaptic organization in LRRTM1/2-DKO mice compared with …
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Figure 1—source data 1
Source data related to Figure 1B, C.
- https://cdn.elifesciences.org/articles/64742/elife-64742-fig1-data1-v1.xlsx
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Figure 1—source data 2
Source data related to Figure 1D, E.
- https://cdn.elifesciences.org/articles/64742/elife-64742-fig1-data2-v1.xlsx
Figure 1—figure supplement 1
LRRTM1/2-DKO generation and confirmation of loss of LRRTM1 and LRRTM2.
(A) LRRTM1/2-DKO mice generation scheme. (B) Western blot images confirming complete loss of LRRTM1 in LRRTM1/2-DKO mice brain homogenate. (C) Western blot images confirming complete loss of LRRTM2 …
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Figure 1—figure supplement 1—source data 1
Figure 1—figure supplement 1 source blots.
- https://cdn.elifesciences.org/articles/64742/elife-64742-fig1-figsupp1-data1-v1.zip
Figure 1—figure supplement 2
Levels of synaptic proteins in crude synaptosomal brain fractions of LRRTM1/2-DKO mice.
(A) LRRTM1/2-DKO mice had unaltered levels of most synaptic proteins in crude synaptosome fractions as compared with wild type (WT) mice. A small but significant increase in synaptophysin levels was …
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Figure 1—figure supplement 2—source data 1
Figure 1—figure supplement 2 source blots.
- https://cdn.elifesciences.org/articles/64742/elife-64742-fig1-figsupp2-data1-v1.zip
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Figure 1—figure supplement 2—source data 2
Figure 1—figure supplement 2 source data.
- https://cdn.elifesciences.org/articles/64742/elife-64742-fig1-figsupp2-data2-v1.xlsx
Figure 1—figure supplement 3
Inhibitory presynapse development is unaltered in CA1 and DG of LRRTM1/2-DKO.
(A and C) High-resolution confocal images revealed no reduction in punctate immunofluorescence for the inhibitory presynapse marker GAD65 in the CA1 dendritic layers and DG molecular layers in …
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Figure 1—figure supplement 3—source data 1
Figure 1—figure supplement 3 source data related to 1B.
- https://cdn.elifesciences.org/articles/64742/elife-64742-fig1-figsupp3-data1-v1.xlsx
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Figure 1—figure supplement 3—source data 2
Figure 1—figure supplement 3 source data related to 1D.
- https://cdn.elifesciences.org/articles/64742/elife-64742-fig1-figsupp3-data2-v1.xlsx
Figure 2
Excitatory synapses are reduced in the CA1 of LRRTM1/2-DKOs.
(A) Golgi staining revealed a reduced density of dendritic spines in CA1 radiatum in the LRRTM1/2-DKO mice as compared with wild-type (WT) mice at 6 weeks postnatal. No differences were observed …
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Figure 2—source data 1
Source data for Figure 2A, B.
- https://cdn.elifesciences.org/articles/64742/elife-64742-fig2-data1-v1.xlsx
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Figure 2—source data 2
Source data for Figure 2D,E.
- https://cdn.elifesciences.org/articles/64742/elife-64742-fig2-data2-v1.xlsx
Figure 3
Excitatory synapse function is differentially altered in the CA1 and DG of LRRTM1/2-DKO mice.
(A and D) Representative mEPSC recordings from wild-type (WT) and LRRTM1/2-DKO hippocampus CA1 pyramidal neurons and DG granule cells. (B and C) Cumulative distributions of mEPSC inter-event …
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Figure 3—source data 1
Source data for Figure 3B, C.
- https://cdn.elifesciences.org/articles/64742/elife-64742-fig3-data1-v1.xlsx
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Figure 3—source data 2
Source data for Figure 3E, F.
- https://cdn.elifesciences.org/articles/64742/elife-64742-fig3-data2-v1.xlsx
Figure 4 with 1 supplement
Impaired hippocampal LTP with hippocampal dependent memory in LRRTM1/2-DKO mice.
(A) The maintenance of long-term potentiation (LTP) is reduced in LRRTM1/2-DKO mouse CA1. Inset: representative fEPSP recordings from wild-type and LRRTM1/2-DKO stratum radiatum CA3-CA1 synapses in …
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Figure 4—source data 1
Source data for Figure 4A, B.
- https://cdn.elifesciences.org/articles/64742/elife-64742-fig4-data1-v1.xlsx
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Figure 4—source data 2
Figure 4D source data.
- https://cdn.elifesciences.org/articles/64742/elife-64742-fig4-data2-v1.xlsx
Figure 4—figure supplement 1
Presynaptic function in CA1 of LRRTM1/2-DKO mice.
(A) Representative traces of five pairs of pulses that were delivered to CA3-CA1 synapses at 20, 50, 100, 200, and 500 ms of LRRTM1/2-DKO and littermate wildtype mice. (B) Summary graph of …
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Figure 4—figure supplement 1—source data 1
Figure 4—figure supplement 1 source data.
- https://cdn.elifesciences.org/articles/64742/elife-64742-fig4-figsupp1-data1-v1.xlsx
Figure 5 with 1 supplement
Excitatory synapse function is impaired in the CA1 of P0-CA1-LRRTM1/2-cDKO mice.
(A) Representative mEPSC recordings from control and P0 LRRTM1/2-cDKO hippocampal CA1 pyramidal neurons. (B and C) Cumulative distributions of mEPSC inter-event intervals (B) and amplitudes (C) in …
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Figure 5—source data 1
Figure 5 source data for 5B and C.
- https://cdn.elifesciences.org/articles/64742/elife-64742-fig5-data1-v1.xlsx
Figure 5—figure supplement 1
Region-specific injection in CA1 of LRRTM1/2floxed/floxed mice in P0 pups to obtain P0-CA1-LRRTM1/2-cDKO mice.
Region specific injection in CA1 of dorsal hippocampus with AAV8-hSYN-Cre-eGFP of P0 LRRTM1/2floxed/floxed mice to obtain a knockout of LRRTM1 and LRRTM2 in CA1 pyramidal neurons. Scale bar is 200 µm.
Figure 6 with 2 supplements
Excitatory presynapse development is unaltered in CA1 dendritic layers and DG molecular layers of LRRTM1/2-cDKO.
(A) Representative image of dorsal hippocampus (injection site) with infected CA1 and dentate gyrus neurons (green) from AAV8-hSYN-Cre-eGFP. Below, zoomed in images of the dorsal CA1 and dentate …
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Figure 6—source data 1
Figure 6 source data for 6C.
- https://cdn.elifesciences.org/articles/64742/elife-64742-fig6-data1-v1.xlsx
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Figure 6—source data 2
Figure 6 source data for 6DG.
- https://cdn.elifesciences.org/articles/64742/elife-64742-fig6-data2-v1.xlsx
Figure 6—figure supplement 1
LRRTM1 and LRRTM2 knockout confirmation.
Representative images of LRRTM1 (A) and LRRTM2 (B) in the dorsal hippocampus showing their expression in the CA1 dendritic layers. (A) LRRTM1 is preferentially expressed in stratum radiatum (SR, …
Figure 6—figure supplement 2
Inhibitory presynapse development is unaltered in CA1 and DG of LRRTM1/2-cDKO mice.
(A and C) High-resolution immunofluorescence images of GAD65 punctate was unaltered in the dendritic layers of CA1 and molecular layers of DG in LRRTM1/2-cDKO mice as compared with control mice. …
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Figure 6—figure supplement 2—source data 1
Figure 6—figure supplement 2 source data.
- https://cdn.elifesciences.org/articles/64742/elife-64742-fig6-figsupp2-data1-v1.xlsx
Figure 7
Excitatory synapse function is impaired in the CA1 and DG of LRRTM1/2-cDKO mice.
(A and D) Representative mEPSC recordings from control and LRRTM1/2-DKO hippocampus CA1 pyramidal neurons and DG granule cells. (B and C) Cumulative distributions of mEPSC inter-event intervals (B) …
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Figure 7—source data 1
Figure 7 source data.
- https://cdn.elifesciences.org/articles/64742/elife-64742-fig7-data1-v1.xlsx
Figure 8 with 1 supplement
Impaired plasticity in dorsal hippocampal CA1 and dentate gyrus regions in LRRTM1/2-cDKO mice.
(A) Epifluoroscent image of dorsal CA1 with stimulating and recording electrodes. Scale bar 50 µm. (B) fEPSP recordings from control and LRRTM1/2-cDKO stratum radiatum CA3-CA1 synapses in acute …
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Figure 8—source data 1
Figure 8 source data.
- https://cdn.elifesciences.org/articles/64742/elife-64742-fig8-data1-v1.xlsx
Figure 8—figure supplement 1
Presynaptic function is unaltered in CA1 of LRRTM1/2-cDKO mice.
(A) Representative traces of five pairs of pulses, which were delivered, to CA3-CA1 synapses at different time intervals of LRRTM1/2-cDKO and control mice. (C) Paired-pulse ratios were unaltered in …
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Figure 8—figure supplement 1—source data 1
Figure 8—figure supplement 1 source data.
- https://cdn.elifesciences.org/articles/64742/elife-64742-fig8-figsupp1-data1-v1.xlsx
Figure 9 with 1 supplement
Contextual memory is impaired in LRRTM1/2-cDKO mice.
(A) Experimental scheme of three-chambered social approach. (B and C) Three chambered social approach test was used to test social memory between controls and LRRTM1/2-cDKO mice. (B) Social novelty …
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Figure 9—source data 1
Figure 9 source data.
- https://cdn.elifesciences.org/articles/64742/elife-64742-fig9-data1-v1.xlsx
Figure 9—figure supplement 1
Behavioral tests performed on LRRTM1/2-cDKO mice showed no differences in motor skills and anxiety-like behavior.
(A) Latency to fall on rotatrod test was comparable between controls and LRRTM1/2-cDKO mice, (Student’s t test, p=0.4202, n=10 for control and n=11 for LRRTM1/2-cDKO). (B–D) Time spent in the inner …
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Figure 9—figure supplement 1—source data 1
Figure 9—figure supplement 1 source data.
- https://cdn.elifesciences.org/articles/64742/elife-64742-fig9-figsupp1-data1-v1.xlsx
