anti lrrtm1 extracellular antibody  (Alomone Labs)


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    Alomone Labs anti lrrtm1 extracellular antibody
    Inhibitory presynapse development is unaltered in CA1 and DG of <t>LRRTM1/2-DKO.</t> ( 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 LRRTM1/2-DKO mice as compared with WT mice at 6 weeks postnatal. Scale bar is 5 µm. ( B and D ) Quantitation of GAD65 punctate integrated intensity per tissue area (Multiple t-test comparing LRRTM1/2-DKO and wild-type mice for CA1 and DG, n=3–5 mice each after averaging data from 6 sections per mouse). Data presented as mean ± SEM. Figure 1—figure supplement 3 source data related to 1B. Figure 1—figure supplement 3 source data related to 1D.
    Anti Lrrtm1 Extracellular Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti lrrtm1 extracellular antibody/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti lrrtm1 extracellular antibody - by Bioz Stars, 2022-11
    94/100 stars

    Images

    1) Product Images from "Distinct but overlapping roles of LRRTM1 and LRRTM2 in developing and mature hippocampal circuits"

    Article Title: Distinct but overlapping roles of LRRTM1 and LRRTM2 in developing and mature hippocampal circuits

    Journal: eLife

    doi: 10.7554/eLife.64742

    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 LRRTM1/2-DKO mice as compared with WT mice at 6 weeks postnatal. Scale bar is 5 µm. ( B and D ) Quantitation of GAD65 punctate integrated intensity per tissue area (Multiple t-test comparing LRRTM1/2-DKO and wild-type mice for CA1 and DG, n=3–5 mice each after averaging data from 6 sections per mouse). Data presented as mean ± SEM. Figure 1—figure supplement 3 source data related to 1B. Figure 1—figure supplement 3 source data related to 1D.
    Figure Legend Snippet: 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 LRRTM1/2-DKO mice as compared with WT mice at 6 weeks postnatal. Scale bar is 5 µm. ( B and D ) Quantitation of GAD65 punctate integrated intensity per tissue area (Multiple t-test comparing LRRTM1/2-DKO and wild-type mice for CA1 and DG, n=3–5 mice each after averaging data from 6 sections per mouse). Data presented as mean ± SEM. Figure 1—figure supplement 3 source data related to 1B. Figure 1—figure supplement 3 source data related to 1D.

    Techniques Used: Immunofluorescence, Marker, Mouse Assay, Quantitation Assay

    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 between genotypes in hippocampal dentate gyrus granule cell medial molecular layer. Scale bar is 1 µm. ( B ) Quantitation of dendritic spine density along CA1 radiatum and dentate gyrus medial molecular layer dendrites (Two-way ANOVA p
    Figure Legend Snippet: 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 between genotypes in hippocampal dentate gyrus granule cell medial molecular layer. Scale bar is 1 µm. ( B ) Quantitation of dendritic spine density along CA1 radiatum and dentate gyrus medial molecular layer dendrites (Two-way ANOVA p

    Techniques Used: Staining, Mouse Assay, Quantitation Assay

    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, top). AAV8-hSYN-Cre-GFP expression in the CA1 of dorsal hippocampus leads to reduced levels of LRRTM1 in the SR (*, bottom). ( B ) LRRTM2 is preferentially expressed in stratum lacunosum moleculare (SLM) of the CA1 and the outer molecular layer of dentate gyrus (top). AAV8-hSYN-Cre-GFP expression in the CA1 of dorsal hippocampus leads to reduced levels of LRRTM2 in the SLM (**) but not in the untargeted dentate gyrus OLM (control). Note that only native GFP is captured in ( A ) and ( B ). Some of the native GFP fluorescence was not captured due to low intensity exposure. Scale bar is 200 µm for all panels except GFP panel for ( A ) which is 100 µm.
    Figure Legend Snippet: 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, top). AAV8-hSYN-Cre-GFP expression in the CA1 of dorsal hippocampus leads to reduced levels of LRRTM1 in the SR (*, bottom). ( B ) LRRTM2 is preferentially expressed in stratum lacunosum moleculare (SLM) of the CA1 and the outer molecular layer of dentate gyrus (top). AAV8-hSYN-Cre-GFP expression in the CA1 of dorsal hippocampus leads to reduced levels of LRRTM2 in the SLM (**) but not in the untargeted dentate gyrus OLM (control). Note that only native GFP is captured in ( A ) and ( B ). Some of the native GFP fluorescence was not captured due to low intensity exposure. Scale bar is 200 µm for all panels except GFP panel for ( A ) which is 100 µm.

    Techniques Used: Knock-Out, Expressing, Fluorescence

    Region-specific injection in CA1 of LRRTM1/2 floxed/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 LRRTM1/2 floxed/floxed mice to obtain a knockout of LRRTM1 and LRRTM2 in CA1 pyramidal neurons. Scale bar is 200 µm.
    Figure Legend Snippet: Region-specific injection in CA1 of LRRTM1/2 floxed/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 LRRTM1/2 floxed/floxed mice to obtain a knockout of LRRTM1 and LRRTM2 in CA1 pyramidal neurons. Scale bar is 200 µm.

    Techniques Used: Injection, Mouse Assay, Knock-Out

    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. Scale bar 5 µm. ( B and D ) Quantitation of GAD65 punctate integrated intensity per tissue area in CA1 and DG (Multiple t-test, n=3 mice each after averaging data from 6 sections per mouse). Data presented as mean ± SEM. Figure 6—figure supplement 2 source data.
    Figure Legend Snippet: 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. Scale bar 5 µm. ( B and D ) Quantitation of GAD65 punctate integrated intensity per tissue area in CA1 and DG (Multiple t-test, n=3 mice each after averaging data from 6 sections per mouse). Data presented as mean ± SEM. Figure 6—figure supplement 2 source data.

    Techniques Used: Mouse Assay, Immunofluorescence, Quantitation Assay

    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 observed (Student’s t-test, *p
    Figure Legend Snippet: 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 observed (Student’s t-test, *p

    Techniques Used: Mouse Assay

    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 intervals ( B ) and amplitudes ( C ) in wild-type and LRRTM1/2-DKO neurons. Insets display mean ± SEM for mEPSC frequency ( B ) and amplitude ( C ) in CA1 pyramidal cells, respectively. Double knockout of Lrrtm1 and Lrrtm2 decreased mEPSC frequency, Student’s t-test, **p=0.0090, without affecting mEPSC amplitude, p=0.6873, n=10–11 neurons per group, three mice each. ( E and F ) Cumulative distributions of mEPSC inter-event intervals ( E ) and amplitudes ( F ) in wild-type and LRRTM1/2-DKO DG granule cells. Insets display mean ± SEM for mEPSC frequency ( E ) and amplitude ( F ) in DG granule cells, respectively. Double knockout of Lrrtm1 and Lrrtm2 increased mEPSC frequency, Student’s t-test, ***p=0.0004, without affecting mEPSC amplitude, p=0.4731, n=11–15 neurons per group, three mice each. Source data for Figure 3B, C . Source data for Figure 3E, F .
    Figure Legend Snippet: 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 intervals ( B ) and amplitudes ( C ) in wild-type and LRRTM1/2-DKO neurons. Insets display mean ± SEM for mEPSC frequency ( B ) and amplitude ( C ) in CA1 pyramidal cells, respectively. Double knockout of Lrrtm1 and Lrrtm2 decreased mEPSC frequency, Student’s t-test, **p=0.0090, without affecting mEPSC amplitude, p=0.6873, n=10–11 neurons per group, three mice each. ( E and F ) Cumulative distributions of mEPSC inter-event intervals ( E ) and amplitudes ( F ) in wild-type and LRRTM1/2-DKO DG granule cells. Insets display mean ± SEM for mEPSC frequency ( E ) and amplitude ( F ) in DG granule cells, respectively. Double knockout of Lrrtm1 and Lrrtm2 increased mEPSC frequency, Student’s t-test, ***p=0.0004, without affecting mEPSC amplitude, p=0.4731, n=11–15 neurons per group, three mice each. Source data for Figure 3B, C . Source data for Figure 3E, F .

    Techniques Used: Mouse Assay, Double Knockout

    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 paired-pulse ratios at various inter-stimulus intervals for wild-type and LRRTM1/2-DKO mice (Multiple t-test, *p > 0.05, n=12 slices per group, three mice each). Figure 4—figure supplement 1 source data.
    Figure Legend Snippet: 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 paired-pulse ratios at various inter-stimulus intervals for wild-type and LRRTM1/2-DKO mice (Multiple t-test, *p > 0.05, n=12 slices per group, three mice each). Figure 4—figure supplement 1 source data.

    Techniques Used: Mouse Assay

    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 ) and amplitudes ( C ) in control and LRRTM1/2-DKO CA1 pyramidal neurons. Insets display mean ± SEM for mEPSC frequency ( B ) and amplitude ( C ) in CA1 pyramidal cells, respectively. Acute conditional deletion of Lrrtm1 and Lrrtm2 in CA1 decreased mEPSC amplitude (Student’s t-test, ***p=0.0004, without affecting mEPSC frequency, p=0.0651. n=16 neurons for three control mice and n=12 neurons for three LRRTM1/2-cDKO mice). ( E and F ) Cumulative distributions of mEPSC inter-event intervals ( E ) and amplitudes ( F ) in control and LRRTM1/2-cDKO neurons. Insets display mean ± SEM for mEPSC frequency ( E ) and amplitude ( F ) in DG granule cells, respectively. Acute conditional deletion of Lrrtm1 and Lrrtm2 in DG decreased mEPSC amplitude (Student’s t-test, ***p=0.0006, without affecting mEPSC frequency, p=0.1444, n=16 neurons for three control mice and n=14 neurons for three LRRTM1/2-cDKO mice). Figure 7 source data.
    Figure Legend Snippet: 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 ) and amplitudes ( C ) in control and LRRTM1/2-DKO CA1 pyramidal neurons. Insets display mean ± SEM for mEPSC frequency ( B ) and amplitude ( C ) in CA1 pyramidal cells, respectively. Acute conditional deletion of Lrrtm1 and Lrrtm2 in CA1 decreased mEPSC amplitude (Student’s t-test, ***p=0.0004, without affecting mEPSC frequency, p=0.0651. n=16 neurons for three control mice and n=12 neurons for three LRRTM1/2-cDKO mice). ( E and F ) Cumulative distributions of mEPSC inter-event intervals ( E ) and amplitudes ( F ) in control and LRRTM1/2-cDKO neurons. Insets display mean ± SEM for mEPSC frequency ( E ) and amplitude ( F ) in DG granule cells, respectively. Acute conditional deletion of Lrrtm1 and Lrrtm2 in DG decreased mEPSC amplitude (Student’s t-test, ***p=0.0006, without affecting mEPSC frequency, p=0.1444, n=16 neurons for three control mice and n=14 neurons for three LRRTM1/2-cDKO mice). Figure 7 source data.

    Techniques Used: Mouse Assay

    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 in LRRTM1/2-DKO mice brain homogenate. Figure 1—figure supplement 1 source blots.
    Figure Legend Snippet: 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 in LRRTM1/2-DKO mice brain homogenate. Figure 1—figure supplement 1 source blots.

    Techniques Used: Mouse Assay, Western Blot

    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 zone versus outer zone of open-field maze (two-way ANOVA, p=0.943), average speed (Student’s t test p=0.6738) and distance travelled (Student’s t test, p=0.9861) was unaltered in LRRTM1/2-cDKO mice, (n=10 for control and n=11 for LRRTM1/2-cDKO). ( E–H ) Anxiety index (time spent in open arms/time spent in closed arms) (Student’s t-test, p=0.0761), number of entries in open arms of elevated plus maze (Student’s t-test, p=0.9018), average speed (Student’s t-test, p=0.6921) and distance travelled (Student’s t-test, p=0.8009) was comparable between controls and LRRTM1/2-cDKO mice (n=10 for control and n=11 for LRRTM1/2-cDKO). Figure 9—figure supplement 1 source data.
    Figure Legend Snippet: 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 zone versus outer zone of open-field maze (two-way ANOVA, p=0.943), average speed (Student’s t test p=0.6738) and distance travelled (Student’s t test, p=0.9861) was unaltered in LRRTM1/2-cDKO mice, (n=10 for control and n=11 for LRRTM1/2-cDKO). ( E–H ) Anxiety index (time spent in open arms/time spent in closed arms) (Student’s t-test, p=0.0761), number of entries in open arms of elevated plus maze (Student’s t-test, p=0.9018), average speed (Student’s t-test, p=0.6921) and distance travelled (Student’s t-test, p=0.8009) was comparable between controls and LRRTM1/2-cDKO mice (n=10 for control and n=11 for LRRTM1/2-cDKO). Figure 9—figure supplement 1 source data.

    Techniques Used: Mouse Assay

    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 LRRTM1/2-cDKO mice CA1 when compared to control mice (Multiple t-test, n=7–8 slices per group, 3–4 mice each group). Figure 8—figure supplement 1 source data.
    Figure Legend Snippet: 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 LRRTM1/2-cDKO mice CA1 when compared to control mice (Multiple t-test, n=7–8 slices per group, 3–4 mice each group). Figure 8—figure supplement 1 source data.

    Techniques Used: Mouse Assay

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    Alomone Labs anti lrrtm1 extracellular antibody
    Inhibitory presynapse development is unaltered in CA1 and DG of <t>LRRTM1/2-DKO.</t> ( 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 LRRTM1/2-DKO mice as compared with WT mice at 6 weeks postnatal. Scale bar is 5 µm. ( B and D ) Quantitation of GAD65 punctate integrated intensity per tissue area (Multiple t-test comparing LRRTM1/2-DKO and wild-type mice for CA1 and DG, n=3–5 mice each after averaging data from 6 sections per mouse). Data presented as mean ± SEM. Figure 1—figure supplement 3 source data related to 1B. Figure 1—figure supplement 3 source data related to 1D.
    Anti Lrrtm1 Extracellular Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti lrrtm1 extracellular antibody/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti lrrtm1 extracellular antibody - by Bioz Stars, 2022-11
    94/100 stars
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    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 LRRTM1/2-DKO mice as compared with WT mice at 6 weeks postnatal. Scale bar is 5 µm. ( B and D ) Quantitation of GAD65 punctate integrated intensity per tissue area (Multiple t-test comparing LRRTM1/2-DKO and wild-type mice for CA1 and DG, n=3–5 mice each after averaging data from 6 sections per mouse). Data presented as mean ± SEM. Figure 1—figure supplement 3 source data related to 1B. Figure 1—figure supplement 3 source data related to 1D.

    Journal: eLife

    Article Title: Distinct but overlapping roles of LRRTM1 and LRRTM2 in developing and mature hippocampal circuits

    doi: 10.7554/eLife.64742

    Figure Lengend Snippet: 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 LRRTM1/2-DKO mice as compared with WT mice at 6 weeks postnatal. Scale bar is 5 µm. ( B and D ) Quantitation of GAD65 punctate integrated intensity per tissue area (Multiple t-test comparing LRRTM1/2-DKO and wild-type mice for CA1 and DG, n=3–5 mice each after averaging data from 6 sections per mouse). Data presented as mean ± SEM. Figure 1—figure supplement 3 source data related to 1B. Figure 1—figure supplement 3 source data related to 1D.

    Article Snippet: The sections were washed with TBSTr (50 mM Tris, pH 7.4, 1.5% NaCl and 0.3% TritonX-100) for 20 min. For immunofluorescence, the slices were then incubated in primary antibodies (LRRTM1 rabbit; Alomone Labs; ANR-141) and (anti-LRRTM2 antibody (510KSCN), described in ) diluted in TBSTr containing 10% normal goat serum for 18–24 hr at 4 °C.

    Techniques: Immunofluorescence, Marker, Mouse Assay, Quantitation Assay

    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 between genotypes in hippocampal dentate gyrus granule cell medial molecular layer. Scale bar is 1 µm. ( B ) Quantitation of dendritic spine density along CA1 radiatum and dentate gyrus medial molecular layer dendrites (Two-way ANOVA p

    Journal: eLife

    Article Title: Distinct but overlapping roles of LRRTM1 and LRRTM2 in developing and mature hippocampal circuits

    doi: 10.7554/eLife.64742

    Figure Lengend Snippet: 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 between genotypes in hippocampal dentate gyrus granule cell medial molecular layer. Scale bar is 1 µm. ( B ) Quantitation of dendritic spine density along CA1 radiatum and dentate gyrus medial molecular layer dendrites (Two-way ANOVA p

    Article Snippet: The sections were washed with TBSTr (50 mM Tris, pH 7.4, 1.5% NaCl and 0.3% TritonX-100) for 20 min. For immunofluorescence, the slices were then incubated in primary antibodies (LRRTM1 rabbit; Alomone Labs; ANR-141) and (anti-LRRTM2 antibody (510KSCN), described in ) diluted in TBSTr containing 10% normal goat serum for 18–24 hr at 4 °C.

    Techniques: Staining, Mouse Assay, Quantitation Assay

    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, top). AAV8-hSYN-Cre-GFP expression in the CA1 of dorsal hippocampus leads to reduced levels of LRRTM1 in the SR (*, bottom). ( B ) LRRTM2 is preferentially expressed in stratum lacunosum moleculare (SLM) of the CA1 and the outer molecular layer of dentate gyrus (top). AAV8-hSYN-Cre-GFP expression in the CA1 of dorsal hippocampus leads to reduced levels of LRRTM2 in the SLM (**) but not in the untargeted dentate gyrus OLM (control). Note that only native GFP is captured in ( A ) and ( B ). Some of the native GFP fluorescence was not captured due to low intensity exposure. Scale bar is 200 µm for all panels except GFP panel for ( A ) which is 100 µm.

    Journal: eLife

    Article Title: Distinct but overlapping roles of LRRTM1 and LRRTM2 in developing and mature hippocampal circuits

    doi: 10.7554/eLife.64742

    Figure Lengend Snippet: 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, top). AAV8-hSYN-Cre-GFP expression in the CA1 of dorsal hippocampus leads to reduced levels of LRRTM1 in the SR (*, bottom). ( B ) LRRTM2 is preferentially expressed in stratum lacunosum moleculare (SLM) of the CA1 and the outer molecular layer of dentate gyrus (top). AAV8-hSYN-Cre-GFP expression in the CA1 of dorsal hippocampus leads to reduced levels of LRRTM2 in the SLM (**) but not in the untargeted dentate gyrus OLM (control). Note that only native GFP is captured in ( A ) and ( B ). Some of the native GFP fluorescence was not captured due to low intensity exposure. Scale bar is 200 µm for all panels except GFP panel for ( A ) which is 100 µm.

    Article Snippet: The sections were washed with TBSTr (50 mM Tris, pH 7.4, 1.5% NaCl and 0.3% TritonX-100) for 20 min. For immunofluorescence, the slices were then incubated in primary antibodies (LRRTM1 rabbit; Alomone Labs; ANR-141) and (anti-LRRTM2 antibody (510KSCN), described in ) diluted in TBSTr containing 10% normal goat serum for 18–24 hr at 4 °C.

    Techniques: Knock-Out, Expressing, Fluorescence

    Region-specific injection in CA1 of LRRTM1/2 floxed/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 LRRTM1/2 floxed/floxed mice to obtain a knockout of LRRTM1 and LRRTM2 in CA1 pyramidal neurons. Scale bar is 200 µm.

    Journal: eLife

    Article Title: Distinct but overlapping roles of LRRTM1 and LRRTM2 in developing and mature hippocampal circuits

    doi: 10.7554/eLife.64742

    Figure Lengend Snippet: Region-specific injection in CA1 of LRRTM1/2 floxed/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 LRRTM1/2 floxed/floxed mice to obtain a knockout of LRRTM1 and LRRTM2 in CA1 pyramidal neurons. Scale bar is 200 µm.

    Article Snippet: The sections were washed with TBSTr (50 mM Tris, pH 7.4, 1.5% NaCl and 0.3% TritonX-100) for 20 min. For immunofluorescence, the slices were then incubated in primary antibodies (LRRTM1 rabbit; Alomone Labs; ANR-141) and (anti-LRRTM2 antibody (510KSCN), described in ) diluted in TBSTr containing 10% normal goat serum for 18–24 hr at 4 °C.

    Techniques: Injection, Mouse Assay, Knock-Out

    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. Scale bar 5 µm. ( B and D ) Quantitation of GAD65 punctate integrated intensity per tissue area in CA1 and DG (Multiple t-test, n=3 mice each after averaging data from 6 sections per mouse). Data presented as mean ± SEM. Figure 6—figure supplement 2 source data.

    Journal: eLife

    Article Title: Distinct but overlapping roles of LRRTM1 and LRRTM2 in developing and mature hippocampal circuits

    doi: 10.7554/eLife.64742

    Figure Lengend Snippet: 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. Scale bar 5 µm. ( B and D ) Quantitation of GAD65 punctate integrated intensity per tissue area in CA1 and DG (Multiple t-test, n=3 mice each after averaging data from 6 sections per mouse). Data presented as mean ± SEM. Figure 6—figure supplement 2 source data.

    Article Snippet: The sections were washed with TBSTr (50 mM Tris, pH 7.4, 1.5% NaCl and 0.3% TritonX-100) for 20 min. For immunofluorescence, the slices were then incubated in primary antibodies (LRRTM1 rabbit; Alomone Labs; ANR-141) and (anti-LRRTM2 antibody (510KSCN), described in ) diluted in TBSTr containing 10% normal goat serum for 18–24 hr at 4 °C.

    Techniques: Mouse Assay, Immunofluorescence, Quantitation Assay

    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 observed (Student’s t-test, *p

    Journal: eLife

    Article Title: Distinct but overlapping roles of LRRTM1 and LRRTM2 in developing and mature hippocampal circuits

    doi: 10.7554/eLife.64742

    Figure Lengend Snippet: 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 observed (Student’s t-test, *p

    Article Snippet: The sections were washed with TBSTr (50 mM Tris, pH 7.4, 1.5% NaCl and 0.3% TritonX-100) for 20 min. For immunofluorescence, the slices were then incubated in primary antibodies (LRRTM1 rabbit; Alomone Labs; ANR-141) and (anti-LRRTM2 antibody (510KSCN), described in ) diluted in TBSTr containing 10% normal goat serum for 18–24 hr at 4 °C.

    Techniques: Mouse Assay

    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 intervals ( B ) and amplitudes ( C ) in wild-type and LRRTM1/2-DKO neurons. Insets display mean ± SEM for mEPSC frequency ( B ) and amplitude ( C ) in CA1 pyramidal cells, respectively. Double knockout of Lrrtm1 and Lrrtm2 decreased mEPSC frequency, Student’s t-test, **p=0.0090, without affecting mEPSC amplitude, p=0.6873, n=10–11 neurons per group, three mice each. ( E and F ) Cumulative distributions of mEPSC inter-event intervals ( E ) and amplitudes ( F ) in wild-type and LRRTM1/2-DKO DG granule cells. Insets display mean ± SEM for mEPSC frequency ( E ) and amplitude ( F ) in DG granule cells, respectively. Double knockout of Lrrtm1 and Lrrtm2 increased mEPSC frequency, Student’s t-test, ***p=0.0004, without affecting mEPSC amplitude, p=0.4731, n=11–15 neurons per group, three mice each. Source data for Figure 3B, C . Source data for Figure 3E, F .

    Journal: eLife

    Article Title: Distinct but overlapping roles of LRRTM1 and LRRTM2 in developing and mature hippocampal circuits

    doi: 10.7554/eLife.64742

    Figure Lengend Snippet: 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 intervals ( B ) and amplitudes ( C ) in wild-type and LRRTM1/2-DKO neurons. Insets display mean ± SEM for mEPSC frequency ( B ) and amplitude ( C ) in CA1 pyramidal cells, respectively. Double knockout of Lrrtm1 and Lrrtm2 decreased mEPSC frequency, Student’s t-test, **p=0.0090, without affecting mEPSC amplitude, p=0.6873, n=10–11 neurons per group, three mice each. ( E and F ) Cumulative distributions of mEPSC inter-event intervals ( E ) and amplitudes ( F ) in wild-type and LRRTM1/2-DKO DG granule cells. Insets display mean ± SEM for mEPSC frequency ( E ) and amplitude ( F ) in DG granule cells, respectively. Double knockout of Lrrtm1 and Lrrtm2 increased mEPSC frequency, Student’s t-test, ***p=0.0004, without affecting mEPSC amplitude, p=0.4731, n=11–15 neurons per group, three mice each. Source data for Figure 3B, C . Source data for Figure 3E, F .

    Article Snippet: The sections were washed with TBSTr (50 mM Tris, pH 7.4, 1.5% NaCl and 0.3% TritonX-100) for 20 min. For immunofluorescence, the slices were then incubated in primary antibodies (LRRTM1 rabbit; Alomone Labs; ANR-141) and (anti-LRRTM2 antibody (510KSCN), described in ) diluted in TBSTr containing 10% normal goat serum for 18–24 hr at 4 °C.

    Techniques: Mouse Assay, Double Knockout

    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 paired-pulse ratios at various inter-stimulus intervals for wild-type and LRRTM1/2-DKO mice (Multiple t-test, *p > 0.05, n=12 slices per group, three mice each). Figure 4—figure supplement 1 source data.

    Journal: eLife

    Article Title: Distinct but overlapping roles of LRRTM1 and LRRTM2 in developing and mature hippocampal circuits

    doi: 10.7554/eLife.64742

    Figure Lengend Snippet: 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 paired-pulse ratios at various inter-stimulus intervals for wild-type and LRRTM1/2-DKO mice (Multiple t-test, *p > 0.05, n=12 slices per group, three mice each). Figure 4—figure supplement 1 source data.

    Article Snippet: The sections were washed with TBSTr (50 mM Tris, pH 7.4, 1.5% NaCl and 0.3% TritonX-100) for 20 min. For immunofluorescence, the slices were then incubated in primary antibodies (LRRTM1 rabbit; Alomone Labs; ANR-141) and (anti-LRRTM2 antibody (510KSCN), described in ) diluted in TBSTr containing 10% normal goat serum for 18–24 hr at 4 °C.

    Techniques: Mouse Assay

    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 ) and amplitudes ( C ) in control and LRRTM1/2-DKO CA1 pyramidal neurons. Insets display mean ± SEM for mEPSC frequency ( B ) and amplitude ( C ) in CA1 pyramidal cells, respectively. Acute conditional deletion of Lrrtm1 and Lrrtm2 in CA1 decreased mEPSC amplitude (Student’s t-test, ***p=0.0004, without affecting mEPSC frequency, p=0.0651. n=16 neurons for three control mice and n=12 neurons for three LRRTM1/2-cDKO mice). ( E and F ) Cumulative distributions of mEPSC inter-event intervals ( E ) and amplitudes ( F ) in control and LRRTM1/2-cDKO neurons. Insets display mean ± SEM for mEPSC frequency ( E ) and amplitude ( F ) in DG granule cells, respectively. Acute conditional deletion of Lrrtm1 and Lrrtm2 in DG decreased mEPSC amplitude (Student’s t-test, ***p=0.0006, without affecting mEPSC frequency, p=0.1444, n=16 neurons for three control mice and n=14 neurons for three LRRTM1/2-cDKO mice). Figure 7 source data.

    Journal: eLife

    Article Title: Distinct but overlapping roles of LRRTM1 and LRRTM2 in developing and mature hippocampal circuits

    doi: 10.7554/eLife.64742

    Figure Lengend Snippet: 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 ) and amplitudes ( C ) in control and LRRTM1/2-DKO CA1 pyramidal neurons. Insets display mean ± SEM for mEPSC frequency ( B ) and amplitude ( C ) in CA1 pyramidal cells, respectively. Acute conditional deletion of Lrrtm1 and Lrrtm2 in CA1 decreased mEPSC amplitude (Student’s t-test, ***p=0.0004, without affecting mEPSC frequency, p=0.0651. n=16 neurons for three control mice and n=12 neurons for three LRRTM1/2-cDKO mice). ( E and F ) Cumulative distributions of mEPSC inter-event intervals ( E ) and amplitudes ( F ) in control and LRRTM1/2-cDKO neurons. Insets display mean ± SEM for mEPSC frequency ( E ) and amplitude ( F ) in DG granule cells, respectively. Acute conditional deletion of Lrrtm1 and Lrrtm2 in DG decreased mEPSC amplitude (Student’s t-test, ***p=0.0006, without affecting mEPSC frequency, p=0.1444, n=16 neurons for three control mice and n=14 neurons for three LRRTM1/2-cDKO mice). Figure 7 source data.

    Article Snippet: The sections were washed with TBSTr (50 mM Tris, pH 7.4, 1.5% NaCl and 0.3% TritonX-100) for 20 min. For immunofluorescence, the slices were then incubated in primary antibodies (LRRTM1 rabbit; Alomone Labs; ANR-141) and (anti-LRRTM2 antibody (510KSCN), described in ) diluted in TBSTr containing 10% normal goat serum for 18–24 hr at 4 °C.

    Techniques: Mouse Assay

    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 in LRRTM1/2-DKO mice brain homogenate. Figure 1—figure supplement 1 source blots.

    Journal: eLife

    Article Title: Distinct but overlapping roles of LRRTM1 and LRRTM2 in developing and mature hippocampal circuits

    doi: 10.7554/eLife.64742

    Figure Lengend Snippet: 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 in LRRTM1/2-DKO mice brain homogenate. Figure 1—figure supplement 1 source blots.

    Article Snippet: The sections were washed with TBSTr (50 mM Tris, pH 7.4, 1.5% NaCl and 0.3% TritonX-100) for 20 min. For immunofluorescence, the slices were then incubated in primary antibodies (LRRTM1 rabbit; Alomone Labs; ANR-141) and (anti-LRRTM2 antibody (510KSCN), described in ) diluted in TBSTr containing 10% normal goat serum for 18–24 hr at 4 °C.

    Techniques: Mouse Assay, Western Blot

    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 zone versus outer zone of open-field maze (two-way ANOVA, p=0.943), average speed (Student’s t test p=0.6738) and distance travelled (Student’s t test, p=0.9861) was unaltered in LRRTM1/2-cDKO mice, (n=10 for control and n=11 for LRRTM1/2-cDKO). ( E–H ) Anxiety index (time spent in open arms/time spent in closed arms) (Student’s t-test, p=0.0761), number of entries in open arms of elevated plus maze (Student’s t-test, p=0.9018), average speed (Student’s t-test, p=0.6921) and distance travelled (Student’s t-test, p=0.8009) was comparable between controls and LRRTM1/2-cDKO mice (n=10 for control and n=11 for LRRTM1/2-cDKO). Figure 9—figure supplement 1 source data.

    Journal: eLife

    Article Title: Distinct but overlapping roles of LRRTM1 and LRRTM2 in developing and mature hippocampal circuits

    doi: 10.7554/eLife.64742

    Figure Lengend Snippet: 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 zone versus outer zone of open-field maze (two-way ANOVA, p=0.943), average speed (Student’s t test p=0.6738) and distance travelled (Student’s t test, p=0.9861) was unaltered in LRRTM1/2-cDKO mice, (n=10 for control and n=11 for LRRTM1/2-cDKO). ( E–H ) Anxiety index (time spent in open arms/time spent in closed arms) (Student’s t-test, p=0.0761), number of entries in open arms of elevated plus maze (Student’s t-test, p=0.9018), average speed (Student’s t-test, p=0.6921) and distance travelled (Student’s t-test, p=0.8009) was comparable between controls and LRRTM1/2-cDKO mice (n=10 for control and n=11 for LRRTM1/2-cDKO). Figure 9—figure supplement 1 source data.

    Article Snippet: The sections were washed with TBSTr (50 mM Tris, pH 7.4, 1.5% NaCl and 0.3% TritonX-100) for 20 min. For immunofluorescence, the slices were then incubated in primary antibodies (LRRTM1 rabbit; Alomone Labs; ANR-141) and (anti-LRRTM2 antibody (510KSCN), described in ) diluted in TBSTr containing 10% normal goat serum for 18–24 hr at 4 °C.

    Techniques: Mouse Assay

    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 LRRTM1/2-cDKO mice CA1 when compared to control mice (Multiple t-test, n=7–8 slices per group, 3–4 mice each group). Figure 8—figure supplement 1 source data.

    Journal: eLife

    Article Title: Distinct but overlapping roles of LRRTM1 and LRRTM2 in developing and mature hippocampal circuits

    doi: 10.7554/eLife.64742

    Figure Lengend Snippet: 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 LRRTM1/2-cDKO mice CA1 when compared to control mice (Multiple t-test, n=7–8 slices per group, 3–4 mice each group). Figure 8—figure supplement 1 source data.

    Article Snippet: The sections were washed with TBSTr (50 mM Tris, pH 7.4, 1.5% NaCl and 0.3% TritonX-100) for 20 min. For immunofluorescence, the slices were then incubated in primary antibodies (LRRTM1 rabbit; Alomone Labs; ANR-141) and (anti-LRRTM2 antibody (510KSCN), described in ) diluted in TBSTr containing 10% normal goat serum for 18–24 hr at 4 °C.

    Techniques: Mouse Assay