Presynaptic PTPσ regulates postsynaptic NMDA receptor function through direct adhesion-independent mechanisms
- Department of Biological Sciences, KAIST, Republic of Korea
- Center for Synaptic Brain Dysfunctions, Institute for Basic Science (IBS), Republic of Korea
- Department of Anatomy and Neurobiology, School of Dentistry, Kyungpook National University, Republic of Korea
- Department of Anatomy and Division of Brain Korea 21, Biomedical Science, College of Medicine, Korea University, Republic of Korea
Figures
Figure 1 with 1 supplement
Normal spontaneous and evoked basal synaptic transmission but suppressed NMDAR-dependent synaptic transmission and plasticity in the Emx1-Cre;Ptprsfl/fl hippocampus.
(A) Normal mEPSCs in CA1 pyramidal neurons from Emx1-Cre;Ptprsfl/fl mice (P18–22). (n = 27 cells from six mice [WT] and 28, 7 [cKO], ns, not significant, Mann-Whitney test). (B) Normal mIPSCs in CA1 …
Figure 1—figure supplement 1
Generation and basic characterization of global Ptprs-mutant mice and Emx1-Cre;Ptprsfl/fl mice.
(A) Conditional KO strategy for the production of Emx1-Cre;Ptprsfl/fl cKO mice. (B) PCR genotyping of Ptprsfl/fl and Emx1-Cre alleles. (C) Reduced levels of PTPσ proteins in hippocampal lysates of …
Figure 2
Deletion of presynaptic, but not postsynaptic, PTPσ suppresses TBS-LTP at SC-CA1 synapses.
(A) Diagram depicting deletion of presynaptic PTPσ in the CA3 region or postsynaptic PTPσ in the CA1 region, induced by local injection of AAV1-hSyn-Cre-eGFP (Cre recombinase fused to EGFP); mice …
Figure 3 with 1 supplement
Re-expression of presynaptic PTP σ rescues postsynaptic LTP in the hippocampus through mechanisms independent of the extracellular region of PTPσ.
(A) Diagram depicting re-expression of WT and mutant PTPσ proteins in the CA3 region of the hippocampus by local injection of AAV(php.eB)-eIF1a-Ptprs (WT and mutants), followed by measurement of …
Figure 3—figure supplement 1
PTPσ adhesions with known postsynaptic partners, and identification of the amino acid residues involved in the interactions.
(A) PTPσ adhesions with known postsynaptic partners based on reported complex structures and molecular modeling of the SALM5-PTPσ complex. (B) Amino acid residues involved in the interactions …
Figure 4 with 1 supplement
Proteins with altered pTyr levels in the Emx1-Cre;Ptprsfl/fl cortex and hippocampus.
(A) Diagram showing procedures for the identification of proteins with altered pTyr levels. (B) A volcano plot showing proteins with significantly up- or downregulated pTyr levels (fold change >1.5, …
Figure 4—figure supplement 1
SynGO analysis of the functions of proteins with altered pTyr Levels in the Emx1-Cre;Ptprsfl/fl cortex and hippocampus.
(A–C) Sunburst plots from SynGO analyses of the function of proteins with altered pTyr levels (total, upregulated, and downregulated) from the Emx1-Cre;Ptprsfl/fl cortex and hippocampus. Note that …
Figure 5 with 1 supplement
Suppressed novelty recognition in Emx1-Cre;Ptprsfl/fl mice.
(A) Impaired novel-object recognition in Emx1-Cre;Ptprsfl/fl mice (2–3 months). (n = 16 mice for WT and cKO, *p<0.05, ns, not significant, Student’s t-test). (B) Normal social approach, but …
Figure 5—figure supplement 1
Emx1-Cre;Ptprsfl/fl mice show moderately changed locomotion and anxiety-like behavior, but normal motor coordination, repetitive behavior, and prepulse inhibition.
(A) Moderately decreased locomotor activity in Emx1-Cre;Ptprsfl/fl mice (2 months) in Laboras cages, representing a familiar environment, as indicated by distance moved. (n = 10 mice [WT] and 15 …
Figure 6
Presynaptic PTPσ-dependent regulation of postsynaptic NMDARs is important for novelty recognition in Emx1-Cre;Ptprsfl/fl mice.
(A) D-cycloserine (DCS) treatment (20 mg/kg; i.p.) rescues social-novelty recognition in Emx1-Cre;Ptprsfl/fl mice (2–3 months) without affecting social approach. Note that DCS has no effect on …
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Gene (Mus musculus) | Ptprs | 19280 in ncbi | For iteration of next studies. | |
| Strain, strain background (Mus musculus; C57BL/6J) | Emx1-Cre;Ptprsfl/fl;Ptprs–l– mice | ES-Cell from KOMP: Ptrpstm1a(KOMP)Mbp | RRID: MGI_5797751 | gKO/cKO mice used in this study |
| Genetic reagent (Mus musculus) | Emx1-Cre | JAX | #005628 | Cre-expressing line used in this study |
| Transfected virus (Mus musculus) | pAAV1-hSyn-Cre-eGFP | Addgene | #105540 | Experimental virus for Cre injection (Figure 2 and Figure 6) |
| Transfected virus (Mus musculus) | pAAV1-hSyn-ΔCre-eGFP | Addgene | #105539 | Control virus for Cre injection (Figure 2 and Figure 6) |
| Transfected construct (Mus musculus) | pAAV-nEFCas9 | Addgene | #87115 | Vector for WT/Mut Ptprs expression (Figure 3). |
| Transfected construct (Mus musculus) | Ptprs WT gene | Li et al., 2015 | Ptprs(meA-/meB-) | Detailed sequence is added in Supplementary file 1. This used for mutagenesis and WT Ptprs rescue injection |
| Transfected construct (Mus musculus) | pAV-pU6-shGluN1#1-GFP | Vigene | #SH836303 | Experimental virus for Cre injection (Figure 6) |
| Transfected construct (Mus musculus) | pAV-pU6-shscrambled-GFP | Vigene | #SH836303 | Control virus for Cre injection (Figure 6) |
| Comparative phosphor-proteomic analysis | PhosphoSCAN service | Cell Signalling Technology | Phospho-tyrosine(pY-1000) | Used in Figure 4 |
| Antibody | anti-PTPσ (Guinea Pig polyclonal) | This paper | #2135 for N-term epitope, #2138 for C-term epitope | WB(1:500) |
| Antibody | anti-PSD93/SynGAP1/NGL3/SALM5/GluA1); (Rabbit,mouse polyclonal) | Home-made; used in previous studies from our group. | #1634(PSD93);#1682(SynGAP1);#2020(NGL3)#1943(SALM50;#1193(GluA1) | WB(1:500) |
| Antibody | anti-tubulin (mouse monoclonal) | DSHB | 12G10 | WB(1:5000) |
| Antibody | anti-GluN1(mouse))/GluN2A(rabbit)) monoclonal | Millipore | Mab363(GluN1);07-632(GluN2A) | WB(1:500) |
| Antibody | anti-GluN2B(mouse)/PSD95(mouse) monoclonal | NeuromAb | 73-101(GluN2B);75-028(PSD95) | WB(1:500) |
| Antibody | anti-Shank3(Rabbit;poly)/Synaptophysin(Mouse;mono) | Santa Cruz | H160(Shank3);D4(Synaptophysin) | WB(1:500) |
| Antibody | anti-linprinα3/caskin1 (Rabbit polyclonal) | Synaptic Systems | 169 102(Liprin α3); 185 003(caskin1) | WB(1:500) |
| Antibody | anti-liprinα2(Rabbit)/Trio(Mouse) polyclonal | Abcam | Ab155411(liprinα2);194365(Trio) | WB(1:500) |
| Antibody | anti-βcatenin(Mouse monoclonal) | BD Science | 610154 | WB(1:500) |
| Antibody | anti-N-Cadherin (Mouse monoclonal) | Thermo | 33–3900 | WB(1:500) |
| Antibody | anti-TrkC (Rabbit monoclonal) | Cell Signaling Technology | 3376 | WB(1:500) |
| Software, algorithm | GraphPad Prism 7.0 | GraphPad | Ver 7.0 | Used for all statistics used in the current study. |
| Software, algorithm | DAVID analysis | David.ncifcrf.gov | DAVID analysis | Used for statistics in proteomics |
Additional files
-
Supplementary file 1
Amino acid sequences of PTPσ mutants used in the present study.
- https://cdn.elifesciences.org/articles/54224/elife-54224-supp1-v2.docx
-
Supplementary file 2
Details of the results of pTyr analysis.
- https://cdn.elifesciences.org/articles/54224/elife-54224-supp2-v2.xlsx
-
Supplementary file 3
Statistical details.
- https://cdn.elifesciences.org/articles/54224/elife-54224-supp3-v2.xlsx
-
Transparent reporting form
- https://cdn.elifesciences.org/articles/54224/elife-54224-transrepform-v2.docx
