title: High-throughput genetic screen for synaptogenic factors: identification of LRP6 as critical for excitatory synapse development.
abstract: Genetic screens in invertebrates have discovered many synaptogenic genes and pathways. However, similar genetic studies have not been possible in mammals. We have optimized an automated high-throughput platform that employs automated liquid handling and imaging of primary mammalian neurons. Using this platform, we have screened 3,200 shRNAs targeting 800 proteins. One of the hits identified was LRP6, a coreceptor for canonical Wnt ligands. LRP6 regulates excitatory synaptogenesis and is selectively localized to excitatory synapses. In vivo knockdown of LRP6 leads to a reduction in the number of functional synapses. Moreover, we show that the canonical Wnt ligand, Wnt8A, promotes synaptogenesis via LRP6. These results provide a proof of principle for using a high-content approach to screen for synaptogenic factors in the mammalian nervous system and identify and characterize a Wnt ligand receptor complex that is critical for the development of functional synapses in vivo.
authors: Department of Neuroscience, Johns Hopkins University School of Medicine, 725 North Wolfe Street, Baltimore, MD 21205, USA.
authors: Kamal
authors: K
authors: Sharma
authors: Department of Physiology, Seoul National University School of Dentistry, Seoul 110-749, South Korea.
authors: Se-Young
authors: SY
authors: Choi
authors: Department of Neuroscience, Johns Hopkins University School of Medicine, 725 North Wolfe Street, Baltimore, MD 21205, USA.
authors: Yong
authors: Y
authors: Zhang
authors: Stanley Center for Psychiatric Research, Broad Institute of Harvard and MIT, 7 Cambridge Center, Cambridge, MA 02142, USA.
authors: Thomas J F
authors: TJ
authors: Nieland
authors: Department of Neuroscience, Johns Hopkins University School of Medicine, 725 North Wolfe Street, Baltimore, MD 21205, USA.
authors: Shunyou
authors: S
authors: Long
authors: Department of Neuroscience, Johns Hopkins University School of Medicine, 725 North Wolfe Street, Baltimore, MD 21205, USA.
authors: Min
authors: M
authors: Li
authors: Department of Neuroscience, Johns Hopkins University School of Medicine, 725 North Wolfe Street, Baltimore, MD 21205, USA. Electronic address: rhuganir@jhmi.edu.
authors: Richard L
authors: RL
authors: Huganir
mesh_terms: D000818:Animals; D003001:Cloning, Molecular / Q000379:methods*; D017362:Cyclic AMP Response Element-Binding Protein / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D057166:High-Throughput Screening Assays / Q000295:instrumentation / Q000379:methods*; D006624:Hippocampus / Q000166:cytology / Q000254:growth & development / Q000378:metabolism; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D008024:Ligands; D060488:Low Density Lipoprotein Receptor-Related Protein-6 / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D009474:Neurons / Q000378:metabolism / Q000502:physiology; D061848:Optical Imaging / Q000379:methods; D011485:Protein Binding; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D051381:Rats; D013569:Synapses / Q000378:metabolism* / Q000502:physiology; D055366:Synaptic Potentials
chemical_list: D017362:Cyclic AMP Response Element-Binding Protein; D036341:Intercellular Signaling Peptides and Proteins; D008024:Ligands; D060488:Low Density Lipoprotein Receptor-Related Protein-6; D034741:RNA, Small Interfering