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Targeted splicing approach for alleviation of a neurexin 1 haploinsufficiency model

Molecular Psychiatry (2025)Cite this article

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Abstract

NRXN1 encoding the synaptic organizing protein neurexin 1 (Nrxn1) is among the strongest risk genes for autism spectrum disorders as well as other neuropsychiatric disorders. The most common contributing mutation is a deletion in one allele. While mice lacking one form of the protein, Nrxn1α, have been characterized, information is lacking on animal models with heterozygous deletion of all isoforms, as well as on therapeutic approaches directly targeting Nrxn1. We report that Nrxn1+/− mice with a deletion affecting all isoforms, α, β and γ, show deficits in excitatory synaptic transmission affecting presynaptic and postsynaptic properties at hippocampal CA3-CA1 synapses, and show increased repetitive behaviors. Based on previous studies indicating that exclusion of the insert at Nrxn1 splice site 5 (S5) boosts synaptic transmission, we tested S5 exclusion as a therapeutic approach. Genetic exclusion of S5 in the remaining Nrxn1 allele alleviated the deficits, restoring miniature excitatory postsynaptic current frequency, paired pulse ratio, AMPA/NMDA ratio, and repetitive behaviors to wild type levels and partially restoring Nrxn1 protein level in Nrxn1ΔS5/- compared to Nrxn1+/− mice. These data suggest that S5 exclusion may be a beneficial therapeutic direction in cases of neuropsychiatric disorders involving NRXN1.

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Fig. 1: Homozygous deletion of Nrxn1 reduces mouse survival.
Fig. 2: Heterozygous deletion of Nrxn1 does not affect dendritic spine density.
Fig. 3: Heterozygous deletion of Nrxn1 disrupts excitatory synaptic transmission with phenotypes normalized by genetic exclusion of S5.
Fig. 4: Presynaptic and postsynaptic properties are disrupted by heterozygous deletion of Nrxn1 and restored by S5 exclusion.
Fig. 5: Heterozygous deletion of Nrxn1 affects repetitive behaviors with phenotypes alleviated by genetic exclusion of S5.
Fig. 6: S5 exclusion partially restores Nrxn1 protein level upon loss of one allele and alters HS valency.

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All data relevant to evaluate the findings of this study are available in the main paper or supplementary material.

References

  1. Sudhof TC. Synaptic neurexin complexes: a molecular code for the logic of neural circuits. Cell. 2017;171:745–69.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Reissner C, Runkel F, Missler M. Neurexins. Genome Biol. 2013;14:213.

    Article  PubMed  PubMed Central  Google Scholar 

  3. Gomez AM, Traunmuller L, Scheiffele P. Neurexins: molecular codes for shaping neuronal synapses. Nat Rev Neurosci. 2021;22:137–51.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Bena F, Bruno DL, Eriksson M, van Ravenswaaij-Arts C, Stark Z, Dijkhuizen T, et al. Molecular and clinical characterization of 25 individuals with exonic deletions of nrxn1 and comprehensive review of the literature. Am J Med Genet B Neuropsychiatr Genet. 2013;162B:388–403.

    Article  PubMed  Google Scholar 

  5. Dabell MP, Rosenfeld JA, Bader P, Escobar LF, El-Khechen D, Vallee SE, et al. Investigation of nrxn1 deletions: clinical and molecular characterization. Am J Med Genet A. 2013;161A:717–31.

    Article  PubMed  Google Scholar 

  6. Castronovo P, Baccarin M, Ricciardello A, Picinelli C, Tomaiuolo P, Cucinotta F, et al. Phenotypic spectrum of nrxn1 mono- and bi-allelic deficiency: a systematic review. Clin Genet. 2020;97:125–37.

    Article  CAS  PubMed  Google Scholar 

  7. Hu Z, Xiao X, Zhang Z, Li M. Genetic insights and neurobiological implications from nrxn1 in neuropsychiatric disorders. Mol Psychiatry. 2019;24:1400–14.

    Article  PubMed  Google Scholar 

  8. Cosemans N, Vandenhove L, Vogels A, Devriendt K, Van Esch H, Van Buggenhout G, et al. The clinical relevance of intragenic nrxn1 deletions. J Med Genet. 2020;57:347–55.

    Article  PubMed  Google Scholar 

  9. Flaherty E, Zhu S, Barretto N, Cheng E, Deans PJM, Fernando MB, et al. Neuronal impact of patient-specific aberrant nrxn1alpha splicing. Nat Genet. 2019;51:1679–90.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Fuccillo MV, Pak C. Copy number variants in neurexin genes: phenotypes and mechanisms. Curr Opin Genet Dev. 2021;68:64–70.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Sebastian R, Jin K, Pavon N, Bansal R, Potter A, Song Y, et al. Schizophrenia-associated nrxn1 deletions induce developmental-timing- and cell-type-specific vulnerabilities in human brain organoids. Nat Commun. 2023;14:3770.

    Article  PubMed  PubMed Central  Google Scholar 

  12. Pak C, Danko T, Zhang Y, Aoto J, Anderson G, Maxeiner S, et al. Human neuropsychiatric disease modeling using conditional deletion reveals synaptic transmission defects caused by heterozygous mutations in nrxn1. Cell Stem Cell. 2015;17:316–28.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Pak C, Danko T, Mirabella VR, Wang J, Liu Y, Vangipuram M, et al. Cross-platform validation of neurotransmitter release impairments in schizophrenia patient-derived nrxn1-mutant neurons. Proc Natl Acad Sci USA. 2021;118:e2025598118.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Etherton MR, Blaiss CA, Powell CM, Sudhof TC. Mouse neurexin-1alpha deletion causes correlated electrophysiological and behavioral changes consistent with cognitive impairments. Proc Natl Acad Sci USA. 2009;106:17998–8003.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Xu B, Ho Y, Fasolino M, Medina J, O’Brien WT, Lamonica JM, et al. Allelic contribution of nrxn1alpha to autism-relevant behavioral phenotypes in mice. PLoS Genet. 2023;19:e1010659.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Grayton HM, Missler M, Collier DA, Fernandes C. Altered social behaviours in neurexin 1alpha knockout mice resemble core symptoms in neurodevelopmental disorders. PLoS ONE. 2013;8:e67114.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Dachtler J, Ivorra JL, Rowland TE, Lever C, Rodgers RJ, Clapcote SJ. Heterozygous deletion of alpha-neurexin i or alpha-neurexin ii results in behaviors relevant to autism and schizophrenia. Behav Neurosci. 2015;129:765–76.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Dai J, Aoto J, Sudhof TC. Alternative splicing of presynaptic neurexins differentially controls postsynaptic nmda and ampa receptor responses. Neuron. 2019;102:993–1008.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Aoto J, Martinelli DC, Malenka RC, Tabuchi K, Sudhof TC. Presynaptic neurexin-3 alternative splicing trans-synaptically controls postsynaptic ampa receptor trafficking. Cell. 2013;154:75–88.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Lu H, Zuo L, Roddick KM, Zhang P, Oku S, Garden J, et al. Alternative splicing and heparan sulfation converge on neurexin-1 to control glutamatergic transmission and autism-related behaviors. Cell Rep. 2023;42:112714.

    Article  CAS  PubMed  Google Scholar 

  21. Zhang P, Lu H, Piexoto RT, Pines MK, Ge Y, Oku S, et al. Heparan sulfate organizes neuronal synapses through neurexin partnerships. Cell. 2018;174:1450–64.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Cardona A, Saalfeld S, Schindelin J, Arganda-Carreras I, Preibisch S, Longair M, et al. Trakem2 software for neural circuit reconstruction. PLoS ONE. 2012;7:e38011.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Blaney CE, Gunn RK, Stover KR, Brown RE. Maternal genotype influences behavioral development of 3xtg-ad mouse pups. Behav Brain Res. 2013;252:40–8.

    Article  CAS  PubMed  Google Scholar 

  24. Dierssen M, Fotaki V, Martinez de Lagran M, Gratacos M, Arbones M, Fillat C, et al. Neurobehavioral development of two mouse lines commonly used in transgenic studies. Pharmacol Biochem Behav. 2002;73:19–25.

    Article  CAS  PubMed  Google Scholar 

  25. Fox WM. Reflex-ontogeny and behavioural development of the mouse. Anim Behav. 1965;13:234–41.

    Article  CAS  PubMed  Google Scholar 

  26. Tremml P, Lipp HP, Muller U, Ricceri L, Wolfer DP. Neurobehavioral development, adult openfield exploration and swimming navigation learning in mice with a modified beta-amyloid precursor protein gene. Behav Brain Res. 1998;95:65–76.

    Article  CAS  PubMed  Google Scholar 

  27. Hill JM, Lim MA, Stone MM Developmental milestones in the newborn mouse. in: Gozes I, Editor. Neuropeptide techniques. Humana Press; Totowa, New Jersey, 2008. p. 131–49.

  28. Friard OGamba M. BORIS: a free, versatile open-source event-logging software for video/audio coding and live observations. Methods Ecol Evol. 2016;7:1325–30.

    Article  Google Scholar 

  29. Kalueff AV, Aldridge JW, LaPorte JL, Murphy DL, Tuohimaa P. Analyzing grooming microstructure in neurobehavioral experiments. Nat Protoc. 2007;2:2538–44.

    Article  CAS  PubMed  Google Scholar 

  30. Deacon RM. Assessing nest building in mice. Nat Protoc. 2006;1:1117–9.

    Article  PubMed  Google Scholar 

  31. Ullrich B, Ushkaryov YA, Sudhof TC. Cartography of neurexins: more than 1000 isoforms generated by alternative splicing and expressed in distinct subsets of neurons. Neuron. 1995;14:497–507.

    Article  CAS  PubMed  Google Scholar 

  32. Uchigashima M, Cheung A, Suh J, Watanabe M, Futai K. Differential expression of neurexin genes in the mouse brain. J Comp Neurol. 2019;527:1940–65.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Trotter JH, Hao J, Maxeiner S, Tsetsenis T, Liu Z, Zhuang X, et al. Synaptic neurexin-1 assembles into dynamically regulated active zone nanoclusters. J Cell Biol. 2019;218:2677–98.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Harris KM, Kater SB. Dendritic spines: cellular specializations imparting both stability and flexibility to synaptic function. Annu Rev Neurosci. 1994;17:341–71.

    Article  CAS  PubMed  Google Scholar 

  35. Gandhi T, Lee CC. Neural mechanisms underlying repetitive behaviors in rodent models of autism spectrum disorders. Front Cell Neurosci. 2020;14:592710.

    Article  PubMed  Google Scholar 

  36. Kim H, Lim CS, Kaang BK. Neuronal mechanisms and circuits underlying repetitive behaviors in mouse models of autism spectrum disorder. Behav Brain Funct. 2016;12:3.

    Article  PubMed  PubMed Central  Google Scholar 

  37. Missler M, Zhang W, Rohlmann A, Kattenstroth G, Hammer RE, Gottmann K, et al. Alpha-neurexins couple ca2+ channels to synaptic vesicle exocytosis. Nature. 2003;423:939–48.

    Article  CAS  PubMed  Google Scholar 

  38. Keck T, Keller GB, Jacobsen RI, Eysel UT, Bonhoeffer T, Hubener M. Synaptic scaling and homeostatic plasticity in the mouse visual cortex in vivo. Neuron. 2013;80:327–34.

    Article  CAS  PubMed  Google Scholar 

  39. Hobbiss AF, Ramiro-Cortes Y, Israely I. Homeostatic plasticity scales dendritic spine volumes and changes the threshold and specificity of hebbian plasticity. iScience. 2018;8:161–74.

    Article  PubMed  PubMed Central  Google Scholar 

  40. Luo F, Sclip A, Jiang M, Sudhof TC. Neurexins cluster ca(2+) channels within the presynaptic active zone. EMBO J. 2020;39:e103208.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Budreck EC, Kwon OB, Jung JH, Baudouin S, Thommen A, Kim HS, et al. Neuroligin-1 controls synaptic abundance of nmda-type glutamate receptors through extracellular coupling. Proc Natl Acad Sci USA. 2013;110:725–30.

    Article  CAS  PubMed  Google Scholar 

  42. Bhouri M, Morishita W, Temkin P, Goswami D, Kawabe H, Brose N, et al. Deletion of LRRTM1 and LRRTM2 in adult mice impairs basal ampa receptor transmission and ltp in hippocampal ca1 pyramidal neurons. Proc Natl Acad Sci USA. 2018;115:E5382–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Dhume SH, Connor SA, Mills F, Tari PK, Au-Yeung SHM, Karimi B, et al. Distinct but overlapping roles of LRRTM1 and LRRTM2 in developing and mature hippocampal circuits. eLife. 2022;11:e64742.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Dai J, Patzke C, Liakath-Ali K, Seigneur E, Sudhof TC. GluD1 is a signal transduction device disguised as an ionotropic receptor. Nature. 2021;595:261–5.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Dai J, Liakath-Ali K, Golf SR, Sudhof TC. Distinct neurexin-cerebellin complexes control ampa- and nmda-receptor responses in a circuit-dependent manner. eLife. 2022;11:e78649.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Nozawa K, Sogabe T, Hayashi A, Motohashi J, Miura E, Arai I, et al. In vivo nanoscopic landscape of neurexin ligands underlying anterograde synapse specification. Neuron. 2022;110:3168–85.

    Article  CAS  PubMed  Google Scholar 

  47. Matsuda K, Budisantoso T, Mitakidis N, Sugaya Y, Miura E, Kakegawa W, et al. Transsynaptic modulation of kainate receptor functions by c1q-like proteins. Neuron. 2016;90:752–67.

    Article  CAS  PubMed  Google Scholar 

  48. Hauser D, Behr K, Konno K, Schreiner D, Schmidt A, Watanabe M, et al. Targeted proteoform mapping uncovers specific neurexin-3 variants required for dendritic inhibition. Neuron. 2022;110:2094–109.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. Mercuri E, Pera MC, Scoto M, Finkel R, Muntoni F. Spinal muscular atrophy - insights and challenges in the treatment era. Nat Rev Neurol. 2020;16:706–15.

    Article  CAS  PubMed  Google Scholar 

  50. Wirth B. Spinal muscular atrophy: in the challenge lies a solution. Trends Neurosci. 2021;44:306–22.

    Article  CAS  PubMed  Google Scholar 

  51. Furlanis E, Traunmuller L, Fucile G, Scheiffele P. Landscape of ribosome-engaged transcript isoforms reveals extensive neuronal-cell-class-specific alternative splicing programs. Nat Neurosci. 2019;22:1709–17.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  52. Lukacsovich D, Winterer J, Que L, Luo W, Lukacsovich T, Foldy C. Single-cell rna-seq reveals developmental origins and ontogenetic stability of neurexin alternative splicing profiles. Cell Rep. 2019;27:3752–9.

    Article  CAS  PubMed  Google Scholar 

  53. Boxer EE, Seng C, Lukacsovich D, Kim J, Schwartz S, Kennedy MJ, et al. Neurexin-3 defines synapse- and sex-dependent diversity of gabaergic inhibition in ventral subiculum. Cell Rep. 2021;37:110098.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

We thank Xiling Zhou for excellent technical assistance, and Sam Barnes, Michael Bellefontaine, Spencer Brown, Val Burdeyny, Emre Fertan, Jessica Garden, Hiyam Gedalia, Elias Habib, Hugh McClelland, Ella Puolitaipale, Qendresa Sahiti, Oliver Schnare, Katie Scott, Aimée Wong, and Jessica Zhang for assistance with behavioural testing. We thank Ed Parker (Vision Core) and Sharmon Knecht, both of the University of Washington, for technical assistance with electron microscopy. Funding was provided by Simons Foundation Autism Research Initiative SFARI 608066 to A.M.C. and R.E.B., Canadian Institutes of Health Research PJT-183943 and FDN-143206 to A.M.C., Natural Sciences and Engineering Research Council of Canada A7441 to R.E.B., National Institutes of Health EYP30-01730 to M. Neitz, a Vanier Canada Graduate Scholarship to H.L., and a Brain Canada’s Shireen and Edna Marcus Excellence Award to H.L.

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Author notes

  1. Kyle M. Roddick

    Present address: Department of Psychology, Mount Allison University, Sackville, NB, E4L 1E4, Canada

  2. Long Zuo

    Present address: Ranomics, Mississauga, ON, L4V 1T4, Canada

Authors and Affiliations

  1. Djavad Mowafaghian Centre for Brain Health and Department of Psychiatry, University of British Columbia, Vancouver, BC, V6T 2B5, Canada

    Hong Lu, Yuan Ge, Long Zuo, Olivia Lam, Klara Marsh & Ann Marie Craig

  2. Department of Psychology and Neuroscience, Dalhousie University, Halifax, NS, B3H 4R2, Canada

    Kyle M. Roddick & Richard E. Brown

  3. Department of Neurosciences, School of Medicine, Case Western Reserve University, Cleveland, OH, 44106, USA

    Peng Zhang

  4. Department of Biological Structure, University of Washington, Seattle, WA, 98195, USA

    Rachel O. L. Wong

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Contributions

The overall project was designed by HL, REB and AMC Experiments were designed, performed, and data analyzed by HL, YG and LZ under the oversight of AMC, by PZ for the knockout validation, by OL and KM under the oversight of HL and ROLW for the SBFSEM, and by KMR under the oversight of REB for the behavioral analyses. The manuscript was written by HL and AMC with input from all authors.

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Correspondence to Ann Marie Craig.

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The authors declare no competing interests.

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All animal procedures were approved by the Animal Care Committee at the University of British Columbia, protocols A16-0086, A20-0095, and A24-0094, and by the University Committee on Laboratory Animals at Dalhousie University, protocols 17–58, 19–072, 19–090, 21–043, 21–079, and 22–063. No human research was performed.

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Lu, H., Roddick, K.M., Ge, Y. et al. Targeted splicing approach for alleviation of a neurexin 1 haploinsufficiency model. Mol Psychiatry (2025). https://doi.org/10.1038/s41380-025-03017-w

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