Regulation of axon guidance and pruning of inappropriate synapses by class 3 semaphorins are key to the development of neural circuits. Collapsin response mediator protein 2 (CRMP2) has been shown to regulate axon guidance by mediating semaphorin 3A (Sema3A) signaling; however, nothing is known about its role in synapse pruning. Here, using newly generated crmp2-/- mice we demonstrate that CRMP2 has a moderate effect on Sema3A-dependent axon guidance in vivo, and its deficiency leads to a mild defect in axon guidance in peripheral nerves and the corpus callosum. Surprisingly, crmp2-/- mice display prominent defects in stereotyped axon pruning in hippocampus and visual cortex and altered dendritic spine remodeling, which is consistent with impaired Sema3F signaling and with models of autism spectrum disorder (ASD). We demonstrate that CRMP2 mediates Sema3F signaling in primary neurons and that crmp2-/- mice display ASD-related social behavior changes in the early postnatal period as well as in adults. Together, we demonstrate that CRMP2 mediates Sema3F-dependent synapse pruning and its dysfunction shares histological and behavioral features of ASD.

Department of Molecular Neurobiology Institute of Physiology of the Czech Academy of Sciences Prague Czech Republic

Department of Neurophysiology of the Memory Institute of Physiology of the Czech Academy of Sciences Prague Czech Republic

Department of Physiology and Pharmacology Sackler Faculty of Medicine Tel Aviv University Tel Aviv Israel

Department of Transgenic Models of Diseases and Czech Centre for Phenogenomics Institute of Molecular Genetics of the Czech Academy of Sciences Prague Czech Republic

Faculty of Science Charles University Prague Prague Czech Republic

German Center for Neurodegenerative Diseases and Munich Cluster for Systems Neurology Munich Germany

Institute of Anatomy and Cell Biology Heidelberg University Heidelberg Germany

Institute of Neuronal Cell Biology Technical University Munich Munich Germany

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Riccomagno MM, Kolodkin AL (2015) Sculpting neural circuits by axon and dendrite pruning. Annu Rev Cell Dev Biol 31: 779–805 PubMed PMC

Johnston MV (2004) Clinical disorders of brain plasticity. Brain Dev 26: 73–80 PubMed

Vanderhaeghen P, Cheng HJ (2010) Guidance molecules in axon pruning and cell death. Cold Spring Harb Perspect Biol 2: a001859 PubMed PMC

Van Battum EY, Brignani S, Pasterkamp RJ (2015) Axon guidance proteins in neurological disorders. Lancet Neurol 14: 532–546 PubMed

Yu F, Schuldiner O (2014) Axon and dendrite pruning in Drosophila . Curr Opin Neurobiol 27: 192–198 PubMed PMC

Bagri A, Cheng HJ, Yaron A, Pleasure SJ, Tessier‐Lavigne M (2003) Stereotyped pruning of long hippocampal axon branches triggered by retraction inducers of the semaphorin family. Cell 113: 285–299 PubMed

Low LK, Liu XB, Faulkner RL, Coble J, Cheng HJ (2008) Plexin signaling selectively regulates the stereotyped pruning of corticospinal axons from visual cortex. Proc Natl Acad Sci USA 105: 8136–8141 PubMed PMC

Goshima Y, Nakamura F, Strittmatter P, Strittmatter SM (1995) Collapsin‐induced growth cone collapse mediated by an intracellular protein related to UNC‐33. Nature 376: 509–514 PubMed

Byk T, Dobransky T, Cifuentes‐Diaz C, Sobel A (1996) Identification and molecular characterization of Unc‐33‐like phosphoprotein (Ulip), a putative mammalian homolog of the axonal guidance‐associated unc‐33 gene product. J Neurosci 16: 688–701 PubMed PMC

Minturn JE, Fryer HJ, Geschwind DH, Hockfield S (1995) TOAD‐64, a gene expressed early in neuronal differentiation in the rat, is related to unc‐33, a C. elegans gene involved in axon outgrowth. J Neurosci 15: 6757–6766 PubMed PMC

Fukata Y, Itoh TJ, Kimura T, Menager C, Nishimura T, Shiromizu T, Watanabe H, Inagaki N, Iwamatsu A, Hotani H et al (2002) CRMP‐2 binds to tubulin heterodimers to promote microtubule assembly. Nat Cell Biol 4: 583–591 PubMed

Uchida Y, Ohshima T, Sasaki Y, Suzuki H, Yanai S, Yamashita N, Nakamura F, Takei K, Ihara Y, Mikoshiba K et al (2005) Semaphorin3A signalling is mediated via sequential Cdk5 and GSK3beta phosphorylation of CRMP2: implication of common phosphorylating mechanism underlying axon guidance and Alzheimer's disease. Genes Cells 10: 165–179 PubMed

Yuasa‐Kawada J, Suzuki R, Kano F, Ohkawara T, Murata M, Noda M (2003) Axonal morphogenesis controlled by antagonistic roles of two CRMP subtypes in microtubule organization. Eur J Neurosci 17: 2329–2343 PubMed

Balastik M, Zhou XZ, Alberich‐Jorda M, Weissova R, Ziak J, Pazyra‐Murphy MF, Cosker KE, Machonova O, Kozmikova I, Chen CH et al (2015) Prolyl isomerase Pin1 regulates axon guidance by stabilizing CRMP2A selectively in distal axons. Cell Rep 13: 812–828 PubMed PMC

Yoshimura T, Kawano Y, Arimura N, Kawabata S, Kikuchi A, Kaibuchi K (2005) GSK‐3beta regulates phosphorylation of CRMP‐2 and neuronal polarity. Cell 120: 137–149 PubMed

Arimura N, Menager C, Kawano Y, Yoshimura T, Kawabata S, Hattori A, Fukata Y, Amano M, Goshima Y, Inagaki M et al (2005) Phosphorylation by Rho kinase regulates CRMP‐2 activity in growth cones. Mol Cell Biol 25: 9973–9984 PubMed PMC

Zhang H, Kang E, Wang Y, Yang C, Yu H, Wang Q, Chen Z, Zhang C, Christian KM, Song H et al (2016) Brain‐specific Crmp2 deletion leads to neuronal development deficits and behavioural impairments in mice. Nat Commun 7: 11773 PubMed PMC

Makihara H, Nakai S, Ohkubo W, Yamashita N, Nakamura F, Kiyonari H, Shioi G, Jitsuki‐Takahashi A, Nakamura H, Tanaka F et al (2016) CRMP1 and CRMP2 have synergistic but distinct roles in dendritic development. Genes Cells 21: 994–1005 PubMed

Nakata K, Ujike H, Sakai A, Takaki M, Imamura T, Tanaka Y, Kuroda S (2003) The human dihydropyrimidinase‐related protein 2 gene on chromosome 8p21 is associated with paranoid‐type schizophrenia. Biol Psychiatry 53: 571–576 PubMed

Liu Y, Pham X, Zhang L, Chen PL, Burzynski G, McGaughey DM, He S, McGrath JA, Wolyniec P, Fallin MD et al (2014) Functional variants in DPYSL2 sequence increase risk of schizophrenia and suggest a link to mTOR signaling. G3 (Bethesda) 5: 61–72 PubMed PMC

Clark D, Dedova I, Cordwell S, Matsumoto I (2006) A proteome analysis of the anterior cingulate cortex gray matter in schizophrenia. Mol Psychiatry 11: 459–470, 423 PubMed

Quach TT, Honnorat J, Kolattukudy PE, Khanna R, Duchemin AM (2015) CRMPs: critical molecules for neurite morphogenesis and neuropsychiatric diseases. Mol Psychiatry 20: 1037–1045 PubMed

Braunschweig D, Krakowiak P, Duncanson P, Boyce R, Hansen RL, Ashwood P, Hertz‐Picciotto I, Pessah IN, Van de Water J (2013) Autism‐specific maternal autoantibodies recognize critical proteins in developing brain. Transl Psychiatry 3: e277. PubMed PMC

De Rubeis S, He X, Goldberg AP, Poultney CS, Samocha K, Cicek AE, Kou Y, Liu L, Fromer M, Walker S et al (2014) Synaptic, transcriptional and chromatin genes disrupted in autism. Nature 515: 209–215 PubMed PMC

Lord C, Elsabbagh M, Baird G, Veenstra‐Vanderweele J (2018) Autism spectrum disorder. Lancet 392: 508–520 PubMed PMC

Bourgeron T (2015) From the genetic architecture to synaptic plasticity in autism spectrum disorder. Nat Rev Neurosci 16: 551–563 PubMed

Kitsukawa T, Shimizu M, Sanbo M, Hirata T, Taniguchi M, Bekku Y, Yagi T, Fujisawa H (1997) Neuropilin‐semaphorin III/D‐mediated chemorepulsive signals play a crucial role in peripheral nerve projection in mice. Neuron 19: 995–1005 PubMed

Taniguchi M, Yuasa S, Fujisawa H, Naruse I, Saga S, Mishina M, Yagi T (1997) Disruption of semaphorin III/D gene causes severe abnormality in peripheral nerve projection. Neuron 19: 519–530 PubMed

Chen H, Bagri A, Zupicich JA, Zou Y, Stoeckli E, Pleasure SJ, Lowenstein DH, Skarnes WC, Chedotal A, Tessier‐Lavigne M (2000) Neuropilin‐2 regulates the development of selective cranial and sensory nerves and hippocampal mossy fiber projections. Neuron 25: 43–56 PubMed

Cheng HJ, Bagri A, Yaron A, Stein E, Pleasure SJ, Tessier‐Lavigne M (2001) Plexin‐A3 mediates semaphorin signaling and regulates the development of hippocampal axonal projections. Neuron 32: 249–263 PubMed

Maimon R, Ionescu A, Bonnie A, Sweetat S, Wald‐Altman S, Inbar S, Gradus T, Trotti D, Weil M, Behar O et al (2018) miR126‐5p downregulation facilitates axon degeneration and NMJ disruption via a non‐cell‐autonomous mechanism in ALS. J Neurosci 38: 5478–5494 PubMed PMC

Brown M, Jacobs T, Eickholt B, Ferrari G, Teo M, Monfries C, Qi RZ, Leung T, Lim L, Hall C (2004) Alpha2‐chimaerin, cyclin‐dependent Kinase 5/p35, and its target collapsin response mediator protein‐2 are essential components in semaphorin 3A‐induced growth‐cone collapse. J Neurosci 24: 8994–9004 PubMed PMC

Zhou J, Wen Y, She L, Sui YN, Liu L, Richards LJ, Poo MM (2013) Axon position within the corpus callosum determines contralateral cortical projection. Proc Natl Acad Sci USA 110: E2714–E2723 PubMed PMC

Wu KY, He M, Hou QQ, Sheng AL, Yuan L, Liu F, Liu WW, Li G, Jiang XY, Luo ZG (2014) Semaphorin 3A activates the guanosine triphosphatase Rab5 to promote growth cone collapse and organize callosal axon projections. Sci Signal 7: ra81 PubMed PMC

Wang CL, Zhang L, Zhou Y, Zhou J, Yang XJ, Duan SM, Xiong ZQ, Ding YQ (2007) Activity‐dependent development of callosal projections in the somatosensory cortex. J Neurosci 27: 11334–11342 PubMed PMC

Penzes P, Cahill ME, Jones KA, VanLeeuwen JE, Woolfrey KM (2011) Dendritic spine pathology in neuropsychiatric disorders. Nat Neurosci 14: 285–293 PubMed PMC

Garey L (2010) When cortical development goes wrong: schizophrenia as a neurodevelopmental disease of microcircuits. J Anat 217: 324–333 PubMed PMC

Liu XB, Low LK, Jones EG, Cheng HJ (2005) Stereotyped axon pruning via plexin signaling is associated with synaptic complex elimination in the hippocampus. J Neurosci 25: 9124–9134 PubMed PMC

Linke R, Pabst T, Frotscher M (1995) Development of the hippocamposeptal projection in the rat. J Comp Neurol 351: 602–616 PubMed

Sahay A, Molliver ME, Ginty DD, Kolodkin AL (2003) Semaphorin 3F is critical for development of limbic system circuitry and is required in neurons for selective CNS axon guidance events. J Neurosci 23: 6671–6680 PubMed PMC

Basarsky TA, Parpura V, Haydon PG (1994) Hippocampal synaptogenesis in cell culture: developmental time course of synapse formation, calcium influx, and synaptic protein distribution. J Neurosci 14: 6402–6411 PubMed PMC

Tran TS, Rubio ME, Clem RL, Johnson D, Case L, Tessier‐Lavigne M, Huganir RL, Ginty DD, Kolodkin AL (2009) Secreted semaphorins control spine distribution and morphogenesis in the postnatal CNS. Nature 462: 1065–1069 PubMed PMC

Yamashita N, Morita A, Uchida Y, Nakamura F, Usui H, Ohshima T, Taniguchi M, Honnorat J, Thomasset N, Takei K et al (2007) Regulation of spine development by semaphorin3A through cyclin‐dependent kinase 5 phosphorylation of collapsin response mediator protein 1. J Neurosci 27: 12546–12554 PubMed PMC

Petanjek Z, Judas M, Simic G, Rasin MR, Uylings HB, Rakic P, Kostovic I (2011) Extraordinary neoteny of synaptic spines in the human prefrontal cortex. Proc Natl Acad Sci USA 108: 13281–13286 PubMed PMC

Bian WJ, Miao WY, He SJ, Qiu Z, Yu X (2015) Coordinated spine pruning and maturation mediated by inter‐spine competition for cadherin/catenin complexes. Cell 162: 808–822 PubMed

Garey LJ, Ong WY, Patel TS, Kanani M, Davis A, Mortimer AM, Barnes TR, Hirsch SR (1998) Reduced dendritic spine density on cerebral cortical pyramidal neurons in schizophrenia. J Neurol Neurosurg Psychiatry 65: 446–453 PubMed PMC

Ey E, Torquet N, Le Sourd AM, Leblond CS, Boeckers TM, Faure P, Bourgeron T (2013) The Autism ProSAP1/Shank2 mouse model displays quantitative and structural abnormalities in ultrasonic vocalisations. Behav Brain Res 256: 677–689 PubMed

Hung AY, Futai K, Sala C, Valtschanoff JG, Ryu J, Woodworth MA, Kidd FL, Sung CC, Miyakawa T, Bear MF et al (2008) Smaller dendritic spines, weaker synaptic transmission, but enhanced spatial learning in mice lacking Shank1. J Neurosci 28: 1697–1708 PubMed PMC

Schmeisser MJ, Ey E, Wegener S, Bockmann J, Stempel AV, Kuebler A, Janssen AL, Udvardi PT, Shiban E, Spilker C et al (2012) Autistic‐like behaviours and hyperactivity in mice lacking ProSAP1/Shank2. Nature 486: 256–260 PubMed

Binder MS, Lugo JN (2017) NS‐Pten knockout mice show sex‐ and age‐specific differences in ultrasonic vocalizations. Brain Behav 7: e00857 PubMed PMC

Reynolds CD, Nolan SO, Jefferson T, Lugo JN (2016) Sex‐specific and genotype‐specific differences in vocalization development in FMR1 knockout mice. NeuroReport 27: 1331–1335 PubMed PMC

Nakamura H, Yamashita N, Kimura A, Kimura Y, Hirano H, Makihara H, Kawamoto Y, Jitsuki‐Takahashi A, Yonezaki K, Takase K et al (2016) Comprehensive behavioral study and proteomic analyses of CRMP2‐deficient mice. Genes Cells 21: 1059–1079 PubMed

Kjelstrup KG, Tuvnes FA, Steffenach HA, Murison R, Moser EI, Moser MB (2002) Reduced fear expression after lesions of the ventral hippocampus. Proc Natl Acad Sci USA 99: 10825–10830 PubMed PMC

Luo L, O'Leary DD (2005) Axon retraction and degeneration in development and disease. Annu Rev Neurosci 28: 127–156 PubMed

Riccomagno MM, Hurtado A, Wang H, Macopson JG, Griner EM, Betz A, Brose N, Kazanietz MG, Kolodkin AL (2012) The RacGAP beta2‐Chimaerin selectively mediates axonal pruning in the hippocampus. Cell 149: 1594–1606 PubMed PMC

Tapia JC, Wylie JD, Kasthuri N, Hayworth KJ, Schalek R, Berger DR, Guatimosim C, Seung HS, Lichtman JW (2012) Pervasive synaptic branch removal in the mammalian neuromuscular system at birth. Neuron 74: 816–829 PubMed

Brill MS, Kleele T, Ruschkies L, Wang M, Marahori NA, Reuter MS, Hausrat TJ, Weigand E, Fisher M, Ahles A et al (2016) Branch‐specific microtubule destabilization mediates axon branch loss during neuromuscular synapse elimination. Neuron 92: 845–856 PubMed PMC

Xu NJ, Henkemeyer M (2009) Ephrin‐B3 reverse signaling through Grb4 and cytoskeletal regulators mediates axon pruning. Nat Neurosci 12: 268–276 PubMed PMC

Cocchi E, Drago A, Serretti A (2016) Hippocampal pruning as a new theory of schizophrenia etiopathogenesis. Mol Neurobiol 53: 2065–2081 PubMed

Sanes JR, Lichtman JW (1999) Development of the vertebrate neuromuscular junction. Annu Rev Neurosci 22: 389–442 PubMed

Venkova K, Christov A, Kamaluddin Z, Kobalka P, Siddiqui S, Hensley K (2014) Semaphorin 3A signaling through neuropilin‐1 is an early trigger for distal axonopathy in the SOD1G93A mouse model of amyotrophic lateral sclerosis. J Neuropathol Exp Neurol 73: 702–713 PubMed PMC

De Winter F, Vo T, Stam FJ, Wisman LA, Bar PR, Niclou SP, van Muiswinkel FL, Verhaagen J (2006) The expression of the chemorepellent Semaphorin 3A is selectively induced in terminal Schwann cells of a subset of neuromuscular synapses that display limited anatomical plasticity and enhanced vulnerability in motor neuron disease. Mol Cell Neurosci 32: 102–117 PubMed

Demyanenko GP, Mohan V, Zhang X, Brennaman LH, Dharbal KE, Tran TS, Manis PB, Maness PF (2014) Neural cell adhesion molecule NrCAM regulates Semaphorin 3F‐induced dendritic spine remodeling. J Neurosci 34: 11274–11287 PubMed PMC

Schwarting GA, Kostek C, Ahmad N, Dibble C, Pays L, Puschel AW (2000) Semaphorin 3A is required for guidance of olfactory axons in mice. J Neurosci 20: 7691–7697 PubMed PMC

Friedel RH, Plump A, Lu X, Spilker K, Jolicoeur C, Wong K, Venkatesh TR, Yaron A, Hynes M, Chen B et al (2005) Gene targeting using a promoterless gene trap vector (“targeted trapping”) is an efficient method to mutate a large fraction of genes. Proc Natl Acad Sci USA 102: 13188–13193 PubMed PMC

Pozas E, Pascual M, Nguyen Ba‐Charvet KT, Guijarro P, Sotelo C, Chedotal A, Del Rio JA, Soriano E (2001) Age‐dependent effects of secreted Semaphorins 3A, 3F, and 3E on developing hippocampal axons: in vitro effects and phenotype of Semaphorin 3A (‐/‐) mice. Mol Cell Neurosci 18: 26–43 PubMed

Schwarz Q, Waimey KE, Golding M, Takamatsu H, Kumanogoh A, Fujisawa H, Cheng HJ, Ruhrberg C (2008) Plexin A3 and plexin A4 convey semaphorin signals during facial nerve development. Dev Biol 324: 1–9 PubMed PMC

Bretin S, Reibel S, Charrier E, Maus‐Moatti M, Auvergnon N, Thevenoux A, Glowinski J, Rogemond V, Premont J, Honnorat J et al (2005) Differential expression of CRMP1, CRMP2A, CRMP2B, and CRMP5 in axons or dendrites of distinct neurons in the mouse brain. J Comp Neurol 486: 1–17 PubMed

Niisato E, Nagai J, Yamashita N, Abe T, Kiyonari H, Goshima Y, Ohshima T (2012) CRMP4 suppresses apical dendrite bifurcation of CA1 pyramidal neurons in the mouse hippocampus. Dev Neurobiol 72: 1447–1457 PubMed

Yamane M, Yamashita N, Hida T, Kamiya Y, Nakamura F, Kolattukudy P, Goshima Y (2017) A functional coupling between CRMP1 and Nav1.7 for retrograde propagation of Semaphorin3A signaling. J Cell Sci 130: 1393–1403 PubMed

Giger RJ, Cloutier JF, Sahay A, Prinjha RK, Levengood DV, Moore SE, Pickering S, Simmons D, Rastan S, Walsh FS et al (2000) Neuropilin‐2 is required in vivo for selective axon guidance responses to secreted semaphorins. Neuron 25: 29–41 PubMed

Ip JP, Fu AK, Ip NY (2014) CRMP2: functional roles in neural development and therapeutic potential in neurological diseases. Neuroscientist 20: 589–598 PubMed

Tobe BTD, Crain AM, Winquist AM, Calabrese B, Makihara H, Zhao WN, Lalonde J, Nakamura H, Konopaske G, Sidor M et al (2017) Probing the lithium‐response pathway in hiPSCs implicates the phosphoregulatory set‐point for a cytoskeletal modulator in bipolar pathogenesis. Proc Natl Acad Sci USA 114: E4462–E4471 PubMed PMC

Dominguez‐Iturza N, Lo AC, Shah D, Armendariz M, Vannelli A, Mercaldo V, Trusel M, Li KW, Gastaldo D, Santos AR et al (2019) The autism‐ and schizophrenia‐associated protein CYFIP1 regulates bilateral brain connectivity and behaviour. Nat Commun 10: 3454 PubMed PMC

Silva AI, Haddon JE, Ahmed Syed Y, Trent S, Lin TE, Patel Y, Carter J, Haan N, Honey RC, Humby T et al (2019) Cyfip1 haploinsufficient rats show white matter changes, myelin thinning, abnormal oligodendrocytes and behavioural inflexibility. Nat Commun 10: 3455 PubMed PMC

Liu X, Malenfant P, Reesor C, Lee A, Hudson ML, Harvard C, Qiao Y, Persico AM, Cohen IL, Chudley AE et al (2011) 2p15‐p16.1 microdeletion syndrome: molecular characterization and association of the OTX1 and XPO1 genes with autism spectrum disorders. Eur J Hum Genet 19: 1264–1270 PubMed PMC

Weimann JM, Zhang YA, Levin ME, Devine WP, Brulet P, McConnell SK (1999) Cortical neurons require Otx1 for the refinement of exuberant axonal projections to subcortical targets. Neuron 24: 819–831 PubMed

Li Z, Jagadapillai R, Gozal E, Barnes G (2019) Deletion of semaphorin 3F in interneurons is associated with decreased GABAergic neurons, autism‐like behavior, and increased oxidative stress cascades. Mol Neurobiol 56: 5520–5538 PubMed PMC

Degano AL, Pasterkamp RJ, Ronnett GV (2009) MeCP2 deficiency disrupts axonal guidance, fasciculation, and targeting by altering Semaphorin 3F function. Mol Cell Neurosci 42: 243–254 PubMed PMC

Cermak T, Doyle EL, Christian M, Wang L, Zhang Y, Schmidt C, Baller JA, Somia NV, Bogdanove AJ, Voytas DF (2011) Efficient design and assembly of custom TALEN and other TAL effector‐based constructs for DNA targeting. Nucleic Acids Res 39: e82 PubMed PMC

Doyle EL, Booher NJ, Standage DS, Voytas DF, Brendel VP, Vandyk JK, Bogdanove AJ (2012) TAL Effector‐Nucleotide Targeter (TALE‐NT) 2.0: tools for TAL effector design and target prediction. Nucleic Acids Res 40: W117–W122 PubMed PMC

Kasparek P, Krausova M, Haneckova R, Kriz V, Zbodakova O, Korinek V, Sedlacek R (2014) Efficient gene targeting of the Rosa26 locus in mouse zygotes using TALE nucleases. FEBS Lett 588: 3982–3988 PubMed

Magiera MM, Bodakuntla S, Ziak J, Lacomme S, Marques Sousa P, Leboucher S, Hausrat TJ, Bosc C, Andrieux A, Kneussel M et al (2018) Excessive tubulin polyglutamylation causes neurodegeneration and perturbs neuronal transport. EMBO J 37: e100440 PubMed PMC

Sherazee N, Alvarez VA (2013) DiOlistics: delivery of fluorescent dyes into cells. Methods Mol Biol 940: 391–400 PubMed PMC

Haddad‐Tovolli R, Szabo NE, Zhou X, Alvarez‐Bolado G (2013) Genetic manipulation of the mouse developing hypothalamus through in utero electroporation. J Vis Exp e50412 PubMed PMC

Ionescu A, Zahavi EE, Gradus T, Ben‐Yaakov K, Perlson E (2016) Compartmental microfluidic system for studying muscle‐neuron communication and neuromuscular junction maintenance. Eur J Cell Biol 95: 69–88 PubMed

Kleele T, Marinkovic P, Williams PR, Stern S, Weigand EE, Engerer P, Naumann R, Hartmann J, Karl RM, Bradke F et al (2014) An assay to image neuronal microtubule dynamics in mice. Nat Commun 5: 4827 PubMed PMC

Brill MS, Marinkovic P, Misgeld T (2013) Sequential photo‐bleaching to delineate single Schwann cells at the neuromuscular junction. J Vis Exp e4460 PubMed PMC

Marinkovic P, Godinho L, Misgeld T (2015) Imaging acute neuromuscular explants from Thy1 mouse lines. Cold Spring Harb Protoc 2015: pdb prot087692 PubMed

Kerschensteiner M, Reuter MS, Lichtman JW, Misgeld T (2008) Ex vivo imaging of motor axon dynamics in murine triangularis sterni explants. Nat Protoc 3: 1645–1653 PubMed PMC

Schindelin J, Arganda‐Carreras I, Frise E, Kaynig V, Longair M, Pietzsch T, Preibisch S, Rueden C, Saalfeld S, Schmid B et al (2012) Fiji: an open‐source platform for biological‐image analysis. Nat Methods 9: 676–682 PubMed PMC

Bures J, Fenton AA, Kaminsky Y, Zinyuk L (1997) Place cells and place navigation. Proc Natl Acad Sci USA 94: 343–350 PubMed PMC

Burghardt NS, Park EH, Hen R, Fenton AA (2012) Adult‐born hippocampal neurons promote cognitive flexibility in mice. Hippocampus 22: 1795–1808 PubMed PMC

Stuchlik A, Petrasek T, Prokopova I, Holubova K, Hatalova H, Vales K, Kubik S, Dockery C, Wesierska M (2013) Place avoidance tasks as tools in the behavioral neuroscience of learning and memory. Physiol Res 62(Suppl 1): S1–S19 PubMed

D'Hooge R, De Deyn PP (2001) Applications of the Morris water maze in the study of learning and memory. Brain Res Brain Res Rev 36: 60–90 PubMed

Wesierska M, Dockery C, Fenton AA (2005) Beyond memory, navigation, and inhibition: behavioral evidence for hippocampus‐dependent cognitive coordination in the rat. J Neurosci 25: 2413–2419 PubMed PMC

Bahník Š (2014) Carousel Maze Manager (Version 0.4.0) Software. Retrieved from https://github.com/bahniks/CM_Manager_0_4_0

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