Molecular reconstitution of functional GABAergic synapses with expression of neuroligin-2 and GABAA receptors

Ning Dong; Jinshun Qi; Gong Chen

doi:10.1016/j.mcn.2007.01.013

Molecular reconstitution of functional GABAergic synapses with expression of neuroligin-2 and GABA_A receptors

Ning Dong
, Jinshun Qi
, Gong Chen

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

64 Scopus citations

Abstract

Trans-synaptic cell adhesion molecules neuroligins and neurexins play an important role in promoting central synapse formation. We report here a molecular reconstruction of functional GABAergic synapses in non-neuronal cells with the coexpression of postsynaptic cell adhesion molecule neuroligin-2 (NL-2) and GABA_A receptors. HEK 293T cells were co-transfected with GABA_A receptors and NL-2 or its homologue neuroligin-1 (NL-1), and then cocultured with hypothalamic cultures which are enriched with GABAergic neurons. Both spontaneous and action potential-evoked GABAergic events were readily detected in HEK 293T cells coexpressing GABA_A receptors with NL-2, but not with NL-1. Aggregating NL-2 with specific antibodies in live cells resulted in coaggregation of GABA_A receptors. Expression of NL-2 in HEK 293T cells also induced stronger GABAergic presynaptic differentiation than that of NL-1 in neuronal cocultures. These results suggest that NL-2 may potentially serve as a central organizer for GABAergic synapse assembly by interacting with both presynaptic neurexins and postsynaptic GABA_A receptors.

Original language	English (US)
Pages (from-to)	14-23
Number of pages	10
Journal	Molecular and Cellular Neuroscience
Volume	35
Issue number	1
DOIs	https://doi.org/10.1016/j.mcn.2007.01.013
State	Published - May 2007

All Science Journal Classification (ASJC) codes

Molecular Biology
Cellular and Molecular Neuroscience
Cell Biology

Access to Document

10.1016/j.mcn.2007.01.013

Cite this

@article{053693fe1e544434b95334246a30b48c,

title = "Molecular reconstitution of functional GABAergic synapses with expression of neuroligin-2 and GABAA receptors",

abstract = "Trans-synaptic cell adhesion molecules neuroligins and neurexins play an important role in promoting central synapse formation. We report here a molecular reconstruction of functional GABAergic synapses in non-neuronal cells with the coexpression of postsynaptic cell adhesion molecule neuroligin-2 (NL-2) and GABAA receptors. HEK 293T cells were co-transfected with GABAA receptors and NL-2 or its homologue neuroligin-1 (NL-1), and then cocultured with hypothalamic cultures which are enriched with GABAergic neurons. Both spontaneous and action potential-evoked GABAergic events were readily detected in HEK 293T cells coexpressing GABAA receptors with NL-2, but not with NL-1. Aggregating NL-2 with specific antibodies in live cells resulted in coaggregation of GABAA receptors. Expression of NL-2 in HEK 293T cells also induced stronger GABAergic presynaptic differentiation than that of NL-1 in neuronal cocultures. These results suggest that NL-2 may potentially serve as a central organizer for GABAergic synapse assembly by interacting with both presynaptic neurexins and postsynaptic GABAA receptors.",

author = "Ning Dong and Jinshun Qi and Gong Chen",

note = "Funding Information: We thank Drs. Bernhard Luscher, Matthew Whim, and June Siqiong Liu for careful reading of our manuscript and insightful comments. We are grateful to Dr. Peter Scheiffele for providing neuroligin constructs and Dr. Luscher for stimulating discussions throughout this project. Dr. Luscher{\textquoteright}s lab also provided substantial material and technical help, including GABA A receptor subunit expression vectors, myc antibody, HEK 293T cells, and the protocol for antibody-induced aggregation and colocalization assays. Min Jiang provided vital help for cell cultures, DNA transfection, plasmid preparation, and fluorescence imaging. This work was supported by National Science Foundation (0236429) and Johnson \& Johnson/Pennsylvania State University Innovative Technology Research Seed Grant. ",

year = "2007",

month = may,

doi = "10.1016/j.mcn.2007.01.013",

language = "English (US)",

volume = "35",

pages = "14--23",

journal = "Molecular and Cellular Neuroscience",

issn = "1044-7431",

publisher = "Academic Press Inc.",

number = "1",

}

TY - JOUR

T1 - Molecular reconstitution of functional GABAergic synapses with expression of neuroligin-2 and GABAA receptors

AU - Dong, Ning

AU - Qi, Jinshun

AU - Chen, Gong

N1 - Funding Information: We thank Drs. Bernhard Luscher, Matthew Whim, and June Siqiong Liu for careful reading of our manuscript and insightful comments. We are grateful to Dr. Peter Scheiffele for providing neuroligin constructs and Dr. Luscher for stimulating discussions throughout this project. Dr. Luscher’s lab also provided substantial material and technical help, including GABA A receptor subunit expression vectors, myc antibody, HEK 293T cells, and the protocol for antibody-induced aggregation and colocalization assays. Min Jiang provided vital help for cell cultures, DNA transfection, plasmid preparation, and fluorescence imaging. This work was supported by National Science Foundation (0236429) and Johnson & Johnson/Pennsylvania State University Innovative Technology Research Seed Grant.

PY - 2007/5

Y1 - 2007/5

N2 - Trans-synaptic cell adhesion molecules neuroligins and neurexins play an important role in promoting central synapse formation. We report here a molecular reconstruction of functional GABAergic synapses in non-neuronal cells with the coexpression of postsynaptic cell adhesion molecule neuroligin-2 (NL-2) and GABAA receptors. HEK 293T cells were co-transfected with GABAA receptors and NL-2 or its homologue neuroligin-1 (NL-1), and then cocultured with hypothalamic cultures which are enriched with GABAergic neurons. Both spontaneous and action potential-evoked GABAergic events were readily detected in HEK 293T cells coexpressing GABAA receptors with NL-2, but not with NL-1. Aggregating NL-2 with specific antibodies in live cells resulted in coaggregation of GABAA receptors. Expression of NL-2 in HEK 293T cells also induced stronger GABAergic presynaptic differentiation than that of NL-1 in neuronal cocultures. These results suggest that NL-2 may potentially serve as a central organizer for GABAergic synapse assembly by interacting with both presynaptic neurexins and postsynaptic GABAA receptors.

AB - Trans-synaptic cell adhesion molecules neuroligins and neurexins play an important role in promoting central synapse formation. We report here a molecular reconstruction of functional GABAergic synapses in non-neuronal cells with the coexpression of postsynaptic cell adhesion molecule neuroligin-2 (NL-2) and GABAA receptors. HEK 293T cells were co-transfected with GABAA receptors and NL-2 or its homologue neuroligin-1 (NL-1), and then cocultured with hypothalamic cultures which are enriched with GABAergic neurons. Both spontaneous and action potential-evoked GABAergic events were readily detected in HEK 293T cells coexpressing GABAA receptors with NL-2, but not with NL-1. Aggregating NL-2 with specific antibodies in live cells resulted in coaggregation of GABAA receptors. Expression of NL-2 in HEK 293T cells also induced stronger GABAergic presynaptic differentiation than that of NL-1 in neuronal cocultures. These results suggest that NL-2 may potentially serve as a central organizer for GABAergic synapse assembly by interacting with both presynaptic neurexins and postsynaptic GABAA receptors.

UR - https://www.scopus.com/pages/publications/34247341381

UR - https://www.scopus.com/pages/publications/34247341381#tab=citedBy

U2 - 10.1016/j.mcn.2007.01.013

DO - 10.1016/j.mcn.2007.01.013

M3 - Article

C2 - 17336090

AN - SCOPUS:34247341381

SN - 1044-7431

VL - 35

SP - 14

EP - 23

JO - Molecular and Cellular Neuroscience

JF - Molecular and Cellular Neuroscience

IS - 1

ER -

Molecular reconstitution of functional GABAergic synapses with expression of neuroligin-2 and GABA_A receptors

Abstract

All Science Journal Classification (ASJC) codes

Access to Document

Other files and links

Fingerprint

Cite this