PMID- 28871037 OWN - NLM STAT- MEDLINE DCOM- 20171023 LR - 20200225 IS - 1529-2401 (Electronic) IS - 0270-6474 (Print) IS - 0270-6474 (Linking) VI - 37 IP - 41 DP - 2017 Oct 11 TI - Identification of Protein Tyrosine Phosphatase Receptor Type O (PTPRO) as a Synaptic Adhesion Molecule that Promotes Synapse Formation. PG - 9828-9843 LID - 10.1523/JNEUROSCI.0729-17.2017 [doi] AB - The proper formation of synapses-specialized unitary structures formed between two neurons-is critical to mediating information flow in the brain. Synaptic cell adhesion molecules (CAMs) are thought to participate in the initiation of the synapse formation process. However, in vivo functional analysis demonstrates that most well known synaptic CAMs regulate synaptic maturation and plasticity rather than synapse formation, suggesting that either CAMs work synergistically in the process of forming synapses or more CAMs remain to be found. By screening for unknown CAMs using a co-culture system, we revealed that protein tyrosine phosphatase receptor type O (PTPRO) is a potent CAM that induces the formation of artificial synapse clusters in co-cultures of human embryonic kidney 293 cells and hippocampal neurons cultured from newborn mice regardless of gender. PTPRO was enriched in the mouse brain and localized to postsynaptic sites at excitatory synapses. The overexpression of PTPRO in cultured hippocampal neurons increased the number of synapses and the frequency of miniature EPSCs (mEPSCs). The knock-down (KD) of PTPRO expression in cultured neurons by short hairpin RNA (shRNA) reduced the number of synapses and the frequencies of the mEPSCs. The effects of shRNA KD were rescued by expressing either full-length PTPRO or a truncated PTPRO lacking the cytoplasmic domain. Consistent with these results, the N-terminal extracellular domain of PTPRO was required for its synaptogenic activity in the co-culture assay. Our data show that PTPRO is a synaptic CAM that serves as a potent initiator of the formation of excitatory synapses.SIGNIFICANCE STATEMENT The formation of synapses is critical for the brain to execute its function and synaptic cell adhesion molecules (CAMs) play essential roles in initiating the formation of synapses. By screening for unknown CAMs using a co-culture system, we revealed that protein tyrosine phosphatase receptor type O (PTPRO) is a potent CAM that induces the formation of artificial synapse clusters. Using loss-of-function and gain-of-function approaches, we show that PTPRO promotes the formation of excitatory synapses. The N-terminal extracellular domain of PTPRO was required for its synaptogenic activity in cultured hippocampal neurons and the co-culture assay. Together, our data show that PTPRO is a synaptic CAM that serves as a potent initiator of synapse formation. CI - Copyright (c) 2017 the authors 0270-6474/17/379828-16$15.00/0. FAU - Jiang, Wei AU - Jiang W AD - Key Laboratory of Cognitive Science, Hubei Key Laboratory of Medical Information Analysis and Tumor Diagnosis and Treatment, Laboratory of Membrane Ion Channels and Medicine, College of Biomedical Engineering, South-Central University for Nationalities, Wuhan 430074, China. AD - State Key Laboratory of Membrane Biology, PKU-IDG/McGovern Institute for Brain Research, School of Life Sciences, Peking University, Beijing 100871, China, and. FAU - Wei, Mengping AU - Wei M AD - State Key Laboratory of Membrane Biology, PKU-IDG/McGovern Institute for Brain Research, School of Life Sciences, Peking University, Beijing 100871, China, and. FAU - Liu, Mengna AU - Liu M AD - State Key Laboratory of Membrane Biology, PKU-IDG/McGovern Institute for Brain Research, School of Life Sciences, Peking University, Beijing 100871, China, and. FAU - Pan, Yunlong AU - Pan Y AD - State Key Laboratory of Membrane Biology, PKU-IDG/McGovern Institute for Brain Research, School of Life Sciences, Peking University, Beijing 100871, China, and. FAU - Cao, Dong AU - Cao D AD - Laboratory Animal Center, Peking University, 5 Yiheyuan Road, Beijing 100871, China. FAU - Yang, Xiaofei AU - Yang X AD - Key Laboratory of Cognitive Science, Hubei Key Laboratory of Medical Information Analysis and Tumor Diagnosis and Treatment, Laboratory of Membrane Ion Channels and Medicine, College of Biomedical Engineering, South-Central University for Nationalities, Wuhan 430074, China, sunlittlefly@hotmail.com ch.zhang@pku.edu.cn. FAU - Zhang, Chen AU - Zhang C AUID- ORCID: 0000-0003-1864-312X AD - State Key Laboratory of Membrane Biology, PKU-IDG/McGovern Institute for Brain Research, School of Life Sciences, Peking University, Beijing 100871, China, and sunlittlefly@hotmail.com ch.zhang@pku.edu.cn. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20170904 PL - United States TA - J Neurosci JT - The Journal of neuroscience : the official journal of the Society for Neuroscience JID - 8102140 RN - 0 (Neural Cell Adhesion Molecules) RN - EC 3.1.3.48 (Ptpro protein, mouse) RN - EC 3.1.3.48 (Receptor-Like Protein Tyrosine Phosphatases, Class 3) SB - IM MH - Animals MH - Animals, Newborn MH - Coculture Techniques MH - Excitatory Postsynaptic Potentials/physiology MH - Gene Knockdown Techniques MH - HEK293 Cells MH - Hippocampus/cytology/growth & development MH - Humans MH - Male MH - Mice MH - Mice, Inbred C57BL MH - Neural Cell Adhesion Molecules/genetics/*physiology MH - Patch-Clamp Techniques MH - Receptor-Like Protein Tyrosine Phosphatases, Class 3/genetics/*physiology MH - Synapses/*physiology PMC - PMC6596594 OTO - NOTNLM OT - PTPRO OT - co-culture OT - electrophysiology OT - morphologic OT - synapse formation OT - synaptic cell adhesion molecules EDAT- 2017/09/06 06:00 MHDA- 2017/10/24 06:00 PMCR- 2018/04/11 CRDT- 2017/09/06 06:00 PHST- 2017/03/22 00:00 [received] PHST- 2017/07/26 00:00 [revised] PHST- 2017/08/22 00:00 [accepted] PHST- 2017/09/06 06:00 [pubmed] PHST- 2017/10/24 06:00 [medline] PHST- 2017/09/06 06:00 [entrez] PHST- 2018/04/11 00:00 [pmc-release] AID - JNEUROSCI.0729-17.2017 [pii] AID - 0729-17 [pii] AID - 10.1523/JNEUROSCI.0729-17.2017 [doi] PST - ppublish SO - J Neurosci. 2017 Oct 11;37(41):9828-9843. doi: 10.1523/JNEUROSCI.0729-17.2017. Epub 2017 Sep 4.