Unbiased discovery of Glypican as a receptor for LRRTM4 in regulating excitatory synapse development

Joris DeWit; Matthew L. O'Sullivan; Jeffrey N. Savas; Giuseppe Condomitti; Max C. Caccese; Kristel M. Vennekens; John R. Yates; Anirvan Ghosh

doi:10.1016/j.neuron.2013.06.049

Unbiased discovery of Glypican as a receptor for LRRTM4 in regulating excitatory synapse development

Joris DeWit^*, Matthew L. O'Sullivan, Jeffrey N. Savas, Giuseppe Condomitti, Max C. Caccese, Kristel M. Vennekens, John R. Yates, Anirvan Ghosh

^*Corresponding author for this work

Neurology

Research output: Contribution to journal › Article › peer-review

125 Scopus citations

Abstract

Leucine-rich repeat (LRR) proteins have recently been identified as important regulators of synapse development and function, but for many LRR proteins the ligand-receptor interactions are not known. Here we identify the heparan sulfate (HS) proteoglycan glypican as a receptor for LRRTM4 using an unbiasedproteomics-based approach. Glypican binds LRRTM4, but not LRRTM2, in an HS-dependent manner. Glypican 4 (GPC4) and LRRTM4 localize to the pre- and postsynaptic membranes of excitatory synapses, respectively. Consistent with a trans-synaptic interaction, LRRTM4 triggers GPC4 clustering in contacting axons and GPC4 induces clustering of LRRTM4 in contacting dendrites in anHS-dependent manner. LRRTM4 positively regulates excitatory synapse development in cultured neurons and invivo, and the synaptogenic activity of LRRTM4 requires the presence of HS on the neuronal surface. Our results identify glypican as an LRRTM4 receptor and indicate that a trans-synaptic glypican-LRRTM4 interaction regulates excitatory synapse development

Original language	English (US)
Pages (from-to)	696-711
Number of pages	16
Journal	Neuron
Volume	79
Issue number	4
DOIs	https://doi.org/10.1016/j.neuron.2013.06.049
State	Published - Aug 21 2013

Funding

We thank the Ghosh laboratory for discussion and Laura DeNardo, Emily Sylwestrak, and Guido David for critical reading of the manuscript. We thank Katie Tiglio, Christine Wu, Christopher Sanchez, Merve Oney, Joseph Antonios, Tev Stachniak, and Stefanie Otto for help with in situ hybridizations, recombinant protein, and virus production and Stéphane Baudouin (Scheiffele laboratory, Biozentrum, University of Basel) for advice on immunohistochemistry. The LRRTM4 monoclonal antibody N205B/22 was developed with the UC Davis/NIH NeuroMab Facility. Mono- and disaccharide analysis of GPC4-Fc was performed by the UCSD Glycotechnology Core. This work was supported by a NARSAD Young Investigator Award from the Brain and Behavior Research Foundation, an ERC Starting Grant (311083) and FWO Odysseus Grant (J.d.W.), National Institute on Aging NRSA Fellowship 1F32AG039127 (J.N.S.), and NIH grants P41 GM103533, R01 MH067880 (J.R.Y.), and R01 NS064124 and NS067216 (A.G.).

ASJC Scopus subject areas

General Neuroscience

Access to Document

10.1016/j.neuron.2013.06.049

Cite this

@article{2d3b7bd47fa841b39569cc9f6f18bee6,

title = "Unbiased discovery of Glypican as a receptor for LRRTM4 in regulating excitatory synapse development",

abstract = "Leucine-rich repeat (LRR) proteins have recently been identified as important regulators of synapse development and function, but for many LRR proteins the ligand-receptor interactions are not known. Here we identify the heparan sulfate (HS) proteoglycan glypican as a receptor for LRRTM4 using an unbiasedproteomics-based approach. Glypican binds LRRTM4, but not LRRTM2, in an HS-dependent manner. Glypican 4 (GPC4) and LRRTM4 localize to the pre- and postsynaptic membranes of excitatory synapses, respectively. Consistent with a trans-synaptic interaction, LRRTM4 triggers GPC4 clustering in contacting axons and GPC4 induces clustering of LRRTM4 in contacting dendrites in anHS-dependent manner. LRRTM4 positively regulates excitatory synapse development in cultured neurons and invivo, and the synaptogenic activity of LRRTM4 requires the presence of HS on the neuronal surface. Our results identify glypican as an LRRTM4 receptor and indicate that a trans-synaptic glypican-LRRTM4 interaction regulates excitatory synapse development",

author = "Joris DeWit and O'Sullivan, {Matthew L.} and Savas, {Jeffrey N.} and Giuseppe Condomitti and Caccese, {Max C.} and Vennekens, {Kristel M.} and Yates, {John R.} and Anirvan Ghosh",

note = "Funding Information: We thank the Ghosh laboratory for discussion and Laura DeNardo, Emily Sylwestrak, and Guido David for critical reading of the manuscript. We thank Katie Tiglio, Christine Wu, Christopher Sanchez, Merve Oney, Joseph Antonios, Tev Stachniak, and Stefanie Otto for help with in situ hybridizations, recombinant protein, and virus production and St{\'e}phane Baudouin (Scheiffele laboratory, Biozentrum, University of Basel) for advice on immunohistochemistry. The LRRTM4 monoclonal antibody N205B/22 was developed with the UC Davis/NIH NeuroMab Facility. Mono- and disaccharide analysis of GPC4-Fc was performed by the UCSD Glycotechnology Core. This work was supported by a NARSAD Young Investigator Award from the Brain and Behavior Research Foundation, an ERC Starting Grant (311083) and FWO Odysseus Grant (J.d.W.), National Institute on Aging NRSA Fellowship 1F32AG039127 (J.N.S.), and NIH grants P41 GM103533, R01 MH067880 (J.R.Y.), and R01 NS064124 and NS067216 (A.G.). ",

year = "2013",

month = aug,

day = "21",

doi = "10.1016/j.neuron.2013.06.049",

language = "English (US)",

volume = "79",

pages = "696--711",

journal = "Neuron",

issn = "0896-6273",

publisher = "Cell Press",

number = "4",

}

TY - JOUR

T1 - Unbiased discovery of Glypican as a receptor for LRRTM4 in regulating excitatory synapse development

AU - DeWit, Joris

AU - O'Sullivan, Matthew L.

AU - Savas, Jeffrey N.

AU - Condomitti, Giuseppe

AU - Caccese, Max C.

AU - Vennekens, Kristel M.

AU - Yates, John R.

AU - Ghosh, Anirvan

N1 - Funding Information: We thank the Ghosh laboratory for discussion and Laura DeNardo, Emily Sylwestrak, and Guido David for critical reading of the manuscript. We thank Katie Tiglio, Christine Wu, Christopher Sanchez, Merve Oney, Joseph Antonios, Tev Stachniak, and Stefanie Otto for help with in situ hybridizations, recombinant protein, and virus production and Stéphane Baudouin (Scheiffele laboratory, Biozentrum, University of Basel) for advice on immunohistochemistry. The LRRTM4 monoclonal antibody N205B/22 was developed with the UC Davis/NIH NeuroMab Facility. Mono- and disaccharide analysis of GPC4-Fc was performed by the UCSD Glycotechnology Core. This work was supported by a NARSAD Young Investigator Award from the Brain and Behavior Research Foundation, an ERC Starting Grant (311083) and FWO Odysseus Grant (J.d.W.), National Institute on Aging NRSA Fellowship 1F32AG039127 (J.N.S.), and NIH grants P41 GM103533, R01 MH067880 (J.R.Y.), and R01 NS064124 and NS067216 (A.G.).

PY - 2013/8/21

Y1 - 2013/8/21

N2 - Leucine-rich repeat (LRR) proteins have recently been identified as important regulators of synapse development and function, but for many LRR proteins the ligand-receptor interactions are not known. Here we identify the heparan sulfate (HS) proteoglycan glypican as a receptor for LRRTM4 using an unbiasedproteomics-based approach. Glypican binds LRRTM4, but not LRRTM2, in an HS-dependent manner. Glypican 4 (GPC4) and LRRTM4 localize to the pre- and postsynaptic membranes of excitatory synapses, respectively. Consistent with a trans-synaptic interaction, LRRTM4 triggers GPC4 clustering in contacting axons and GPC4 induces clustering of LRRTM4 in contacting dendrites in anHS-dependent manner. LRRTM4 positively regulates excitatory synapse development in cultured neurons and invivo, and the synaptogenic activity of LRRTM4 requires the presence of HS on the neuronal surface. Our results identify glypican as an LRRTM4 receptor and indicate that a trans-synaptic glypican-LRRTM4 interaction regulates excitatory synapse development

AB - Leucine-rich repeat (LRR) proteins have recently been identified as important regulators of synapse development and function, but for many LRR proteins the ligand-receptor interactions are not known. Here we identify the heparan sulfate (HS) proteoglycan glypican as a receptor for LRRTM4 using an unbiasedproteomics-based approach. Glypican binds LRRTM4, but not LRRTM2, in an HS-dependent manner. Glypican 4 (GPC4) and LRRTM4 localize to the pre- and postsynaptic membranes of excitatory synapses, respectively. Consistent with a trans-synaptic interaction, LRRTM4 triggers GPC4 clustering in contacting axons and GPC4 induces clustering of LRRTM4 in contacting dendrites in anHS-dependent manner. LRRTM4 positively regulates excitatory synapse development in cultured neurons and invivo, and the synaptogenic activity of LRRTM4 requires the presence of HS on the neuronal surface. Our results identify glypican as an LRRTM4 receptor and indicate that a trans-synaptic glypican-LRRTM4 interaction regulates excitatory synapse development

UR - http://www.scopus.com/inward/record.url?scp=84882645603&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84882645603&partnerID=8YFLogxK

U2 - 10.1016/j.neuron.2013.06.049

DO - 10.1016/j.neuron.2013.06.049

M3 - Article

C2 - 23911103

AN - SCOPUS:84882645603

SN - 0896-6273

VL - 79

SP - 696

EP - 711

JO - Neuron

JF - Neuron

IS - 4

ER -

Unbiased discovery of Glypican as a receptor for LRRTM4 in regulating excitatory synapse development

Abstract

Funding

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this