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Genetic Targeting of NRXN2 in Mice Unveils Role in Excitatory Cortical Synapse Function and Social Behaviors

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Genetic Targeting of NRXN2 in Mice Unveils Role in Excitatory Cortical Synapse Function and Social Behaviors King’s Research Portal DOI: 10.3389/fnsyn.2015.00003 Document Version Publisher's PDF, also known as Version of record Link to publication record in King's Research Portal Citation for published version (APA): Born, G., Grayton, H. M., Langhorst, H., Dudanova, I., Rohlmann, A., Woodward, B. W., Collier, D. A., Fernandes, C., & Missler, M. (2015). Genetic targeting of NRXN2 in mice unveils role in excitatory cortical synapse function and social behaviors. Frontiers in Synaptic Neuroscience, 7(0), [3]. https://doi.org/10.3389/fnsyn.2015.00003 Citing this paper Please note that where the full-text provided on King's Research Portal is the Author Accepted Manuscript or Post-Print version this may differ from the final Published version. If citing, it is advised that you check and use the publisher's definitive version for pagination, volume/issue, and date of publication details. 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Download date: 27. Sep. 2021 ORIGINAL RESEARCH ARTICLE published: 19 February 2015 SYNAPTIC NEUROSCIENCE doi: 10.3389/fnsyn.2015.00003 Genetic targeting of NRXN2 in mice unveils role in excitatory cortical synapse function and social behaviors Gesche Born 1‡, Hannah M. Grayton 2,3‡, Hanna Langhorst 1†, Irina Dudanova 1†, Astrid Rohlmann 1, Benjamin W. Woodward 2, David A. Collier 2,3, Cathy Fernandes 2‡ and Markus Missler 1,4*‡ 1 Institute of Anatomy and Molecular Neurobiology, Westfälische Wilhelms-University, Münster, Germany 2 Social, Genetic and Developmental Psychiatry Centre, Institute of Psychiatry, Psychology and Neuroscience, King’s College London, London, UK 3 Discovery Neuroscience Research, Eli Lilly and Company Ltd., Surrey, UK 4 Cluster of Excellence EXC 1003, Cells in Motion, Münster, Germany Edited by: Human genetics has identified rare copy number variations and deleterious mutations Volkmar Lessmann, for all neurexin genes (NRXN1-3) in patients with neurodevelopmental diseases, and Otto-von-Guericke University, electrophysiological recordings in animal brains have shown that Nrxns are important Germany for synaptic transmission. While several mouse models for Nrxn1α inactivation have Reviewed by: Simon Rumpel, Research Institute previously been studied for behavioral changes, very little information is available for other of Molecular Pathology, Austria variants. Here, we validate that mice lacking Nrxn2α exhibit behavioral abnormalities, Thomas Endres, Otto-von-Guericke characterized by social interaction deficits and increased anxiety-like behavior, which University, Germany partially overlap, partially differ from Nrxn1α mutant behaviors. Using patch-clamp *Correspondence: recordings in Nrxn2α knockout brains, we observe reduced spontaneous transmitter Markus Missler, Institute of Anatomy and Molecular release at excitatory synapses in the neocortex. We also analyse at this cellular level Neurobiology, Westfälische anovelNRXN2 mouse model that carries a combined deletion of Nrxn2α and Nrxn2β. Wilhelms-University, Vesaliusweg Electrophysiological analysis of this Nrxn2-mutant mouse shows surprisingly similar 2-4, 48149 Münster, Germany defects of excitatory release to Nrxn2α, indicating that the β-variant of Nrxn2 has no e-mail: Markus.Missler@ uni-muenster.de strong function in basic transmission at these synapses. Inhibitory transmission as †Present address: well as synapse densities and ultrastructure remain unchanged in the neocortex of Hanna Langhorst and Irina both models. Furthermore, at Nrxn2α and Nrxn2-mutant excitatory synapses we find Dudanova, Max Planck Institute of an altered facilitation and N-methyl-D-aspartate receptor (NMDAR) function because Neurobiology, Martinsried, Germany NMDAR-dependent decay time and NMDAR-mediated responses are reduced. As Nrxn (I.D.); Max-Delbrück-Centrum für Molekulare Medizin, Berlin, can indirectly be linked to NMDAR via neuroligin and PSD-95, the trans-synaptic nature Germany (H.L.) of this complex may help to explain occurrence of presynaptic and postsynaptic effects. ‡These authors have contributed Since excitatory/inhibitory imbalances and impairment of NMDAR function are alledged equally to this work. to have a role in autism and schizophrenia, our results support the idea of a related pathomechanism in these disorders. Keywords: vesicle release, synaptic plasticity, cognition, electron microscopy, AMPA receptor, NMDA receptor, anxiety-like behavior, social interaction INTRODUCTION isoforms, from independent promoters (Tabuchi and Sudhof, Synapse function and plasticity in the brain depend on the coor- 2002; Runkel et al., 2013). These are then alternatively spliced dinated action of molecular machineries across the synaptic cleft. at five, possibly six, conserved sites (Tabuchi and Sudhof, 2002; Presynaptic neurexins (Nrxns) form trans-synaptic complexes Treutlein et al., 2014). α-Nrxns encode six LNS domains with with postsynaptic neuroligins and other partners that are suited to three interspersed EGF-like domains, while β-Nrxns contain a mediate such function (Missler et al., 2012; Reissner et al., 2013). single LNS-domain that is identical to the sixth LNS-domain Consistent with this role, an association has been established of the corresponding α-Nrxn, and mediates binding, for exam- between copy number variations or hemizygous exonic deletions ple, to neuroligins (Ichtchenko et al., 1995; Boucard et al., 2005; within the NRXN1 gene (2p16.3) and neurodevelopmental dis- Reissner et al., 2008). Studies of knockout mice have revealed orders, including autism, schizophrenia and intellectual disability essential functions for α-Nrxns and neuroligins at excitatory and (Feng et al., 2006; Szatmari et al., 2007; Kim et al., 2008; Kirov inhibitory synapses (Missler et al., 2003; Varoqueaux et al., 2006). et al., 2008; Yan et al., 2008; Zahir et al., 2008; Rujescu et al., 2009; Multiple knockout of all three α-Nrxns is lethal at birth due to a Zweier et al., 2009; Ching et al., 2010; Voineskos et al., 2011; Yue dramatic impairment of Ca2+-dependent vesicle release (Missler et al., 2011; Buxbaum et al., 2012; Camacho-Garcia et al., 2012; et al., 2003; Kattenstroth et al., 2004; Zhang et al., 2005), but even Liu et al., 2012; Hoeffding et al., 2014). removal of a single Nrxn1α isoform reduces glutamatergic trans- Vertebrates contain three NRXN genes (NRXN1-3) that mission (Missler et al., 2003; Etherton et al., 2009). While levels each produce longer α-Nrxn and shorter β-Nrxn extracellular of β-Nrxns appeared unchanged in α-Nrxns KO mice (Missler Frontiers in Synaptic Neuroscience www.frontiersin.org February 2015 | Volume 7 | Article 3 | 1 Born et al. Abnormalities in neurexin-2 mouse mutants et al., 2003), no information of deletion of β-Nrxns in mice is ethical review panel of King’s College, and U.K. Home Office available yet. Animals Scientific Procedures Act 1986 (license PPL:70/7184). In studies on autism spectrum disorders, which are character- Other experiments were approved by the Landesamt für Natur, ized clinically by impairments in reciprocal social interactions and Umwelt und Verbraucherschutz NRW, Germany (license 84- communication as well as stereotyped behaviors, rare heterozy- 02.05.20.11.209). gous deletions in the NRXN1 gene are considered as causative variants (Delorme et al., 2013). Consequently, several studies have Combined Nrxn2 mouse model addressed higher brain functions in NRXN1 mouse models by To delete the α-andβ-variants of the Nrxn2 gene together, part behavioral profiling (Ey et al., 2011). Nrxn1α homozygote KO of exon 23 (Tabuchi and Sudhof, 2002) was deleted, and the mice display a decrease in social investigation and an increase gene interrupted between splice insert #5 into the 3UTR. Mutant in aggressive behavior as well as reduced locomotor activity mice were originally generated in Thomas C. Südhof’s laboratory in novel environments (Grayton et al., 2013), impairments in (Stanford University, USA), which will also provide additional prepulse inhibition of the startle response, deficits in nest build- analyses at a later time point. Genotyping was performed by PCR ing activities, and an improvement in motor learning (Etherton with oligonucleotide primer combinations: 5-AGC CCC GAC et al., 2009). Furthermore, Nrxn1α heterozygote mice have been CCA ACC TCA GGA CAG A -3 vs. 5-GGT AGG GAC AAG AGA shown to have sex-dependent increases in response to novelty and CAG CAA-3 (WT allele, 1 000 bp product), and 5-CGC CGC accelerated habituation to novel environments (Laarakker et al., TCC CGA TTC GCA GCG CAT-3 vs. 5-GGT AGG GAC AAG 2012). AGA CAG CAA-3 (KO allele, 700 bp product). Although genetic
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