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Open Access 01-12-2010 | Research

Haploinsufficiency of the autism-associated Shank3 gene leads to deficits in synaptic function, social interaction, and social communication

Authors: Ozlem Bozdagi, Takeshi Sakurai, Danae Papapetrou, Xiaobin Wang, Dara L Dickstein, Nagahide Takahashi, Yuji Kajiwara, Mu Yang, Adam M Katz, Maria Luisa Scattoni, Mark J Harris, Roheeni Saxena, Jill L Silverman, Jacqueline N Crawley, Qiang Zhou, Patrick R Hof, Joseph D Buxbaum

Published in: Molecular Autism | Issue 1/2010

Abstract

Background

SHANK3 is a protein in the core of the postsynaptic density (PSD) and has a critical role in recruiting many key functional elements to the PSD and to the synapse, including components of α-amino-3-hydroxyl-5-methyl-4-isoxazole-propionic acid (AMPA), metabotropic glutamate (mGlu) and N-methyl-D-aspartic acid (NMDA) glutamate receptors, as well as cytoskeletal elements. Loss of a functional copy of the SHANK3 gene leads to the neurobehavioral manifestations of 22q13 deletion syndrome and/or to autism spectrum disorders. The goal of this study was to examine the effects of haploinsufficiency of full-length Shank3 in mice, focusing on synaptic development, transmission and plasticity, as well as on social behaviors, as a model for understanding SHANK3 haploinsufficiency in humans.

Methods

We used mice with a targeted disruption of Shank3 in which exons coding for the ankyrin repeat domain were deleted and expression of full-length Shank3 was disrupted. We studied synaptic transmission and plasticity by multiple methods, including patch-clamp whole cell recording, two-photon time-lapse imaging and extracellular recordings of field excitatory postsynaptic potentials. We also studied the density of GluR1-immunoreactive puncta in the CA1 stratum radiatum and carried out assessments of social behaviors.

Results

In Shank3 heterozygous mice, there was reduced amplitude of miniature excitatory postsynaptic currents from hippocampal CA1 pyramidal neurons and the input-output (I/O) relationship at Schaffer collateral-CA1 synapses in acute hippocampal slices was significantly depressed; both of these findings indicate a reduction in basal neurotransmission. Studies with specific inhibitors demonstrated that the decrease in basal transmission reflected reduced AMPA receptor-mediated transmission. This was further supported by the observation of reduced numbers of GluR1-immunoreactive puncta in the stratum radiatum. Long-term potentiation (LTP), induced either with θ-burst pairing (TBP) or high-frequency stimulation, was impaired in Shank3 heterozygous mice, with no significant change in long-term depression (LTD). In concordance with the LTP results, persistent expansion of spines was observed in control mice after TBP-induced LTP; however, only transient spine expansion was observed in Shank3 heterozygous mice. Male Shank3 heterozygotes displayed less social sniffing and emitted fewer ultrasonic vocalizations during interactions with estrus female mice, as compared to wild-type littermate controls.

Conclusions

We documented specific deficits in synaptic function and plasticity, along with reduced reciprocal social interactions in Shank3 heterozygous mice. Our results are consistent with altered synaptic development and function in Shank3 haploinsufficiency, highlighting the importance of Shank3 in synaptic function and supporting a link between deficits in synapse function and neurodevelopmental disorders. The reduced glutamatergic transmission that we observed in the Shank3 heterozygous mice represents an interesting therapeutic target in Shank3-haploinsufficiency syndromes.

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Naisbitt S, Kim E, Tu JC, Xiao B, Sala C, Valtschanoff J, Weinberg RJ, Worley PF, Sheng M: Shank, a novel family of postsynaptic density proteins that binds to the NMDA receptor/PSD-95/GKAP complex and cortactin. Neuron. 1999, 23: 569-582. 10.1016/S0896-6273(00)80809-0.CrossRefPubMed

Sheng M: Excitatory synapses. Glutamate receptors put in their place. Nature. 1997, 386: 221-223. 10.1038/386221a0.CrossRefPubMed

Sheng M, Kim E: The Shank family of scaffold proteins. J Cell Sci. 2000, 113 (Pt 11): 1851-1856.PubMed

Yao I, Iida J, Nishimura W, Hata Y: Synaptic localization of SAPAP1, a synaptic membrane-associated protein. Genes Cells. 2003, 8: 121-129. 10.1046/j.1365-2443.2003.00622.x.CrossRefPubMed

Zitzer H, Richter D, Kreienkamp HJ: Agonist-dependent interaction of the rat somatostatin receptor subtype 2 with cortactin-binding protein 1. J Biol Chem. 1999, 274: 18153-18156. 10.1074/jbc.274.26.18153.CrossRefPubMed

Kreienkamp HJ, Soltau M, Richter D, Bockers T: Interaction of G-protein-coupled receptors with synaptic scaffolding proteins. Biochem Soc Trans. 2002, 30: 464-468. 10.1042/BST0300464.CrossRefPubMed

Tu JC, Xiao B, Naisbitt S, Yuan JP, Petralia RS, Brakeman P, Doan A, Aakalu VK, Lanahan AA, Sheng M, Worley PF: Coupling of mGluR/Homer and PSD-95 complexes by the Shank family of postsynaptic density proteins. Neuron. 1999, 23: 583-592. 10.1016/S0896-6273(00)80810-7.CrossRefPubMed

Kim JH, Kim JH, Yang E, Park JH, Yu YS, Kim KW: Shank 2 expression coincides with neuronal differentiation in the developing retina. Exp Mol Med. 2009, 41: 236-242. 10.3858/emm.2009.41.4.026.PubMedCentralCrossRefPubMed

Qualmann B, Boeckers TM, Jeromin M, Gundelfinger ED, Kessels MM: Linkage of the actin cytoskeleton to the postsynaptic density via direct interactions of Abp1 with the ProSAP/Shank family. J Neurosci. 2004, 24: 2481-2495. 10.1523/JNEUROSCI.5479-03.2004.CrossRefPubMed

10.

Haeckel A, Ahuja R, Gundelfinger ED, Qualmann B, Kessels MM: The actin-binding protein Abp1 controls dendritic spine morphology and is important for spine head and synapse formation. J Neurosci. 2008, 28: 10031-10044. 10.1523/JNEUROSCI.0336-08.2008.CrossRefPubMed

11.

Du Y, Weed SA, Xiong WC, Marshall TD, Parsons JT: Identification of a novel cortactin SH3 domain-binding protein and its localization to growth cones of cultured neurons. Mol Cell Biol. 1998, 18: 5838-5851.PubMedCentralCrossRefPubMed

12.

Cingolani LA, Goda Y: Actin in action: the interplay between the actin cytoskeleton and synaptic efficacy. Nat Rev Neurosci. 2008, 9: 344-356. 10.1038/nrn2373.CrossRefPubMed

13.

Sheng M, Kim MJ: Postsynaptic signaling and plasticity mechanisms. Science. 2002, 298: 776-780. 10.1126/science.1075333.CrossRefPubMed

14.

Vanderklish PW, Krushel LA, Holst BH, Gally JA, Crossin KL, Edelman GM: Marking synaptic activity in dendritic spines with a calpain substrate exhibiting fluorescence resonance energy transfer. Proc Natl Acad Sci USA. 2000, 97: 2253-2258. 10.1073/pnas.040565597.PubMedCentralCrossRefPubMed

15.

Lim S, Naisbitt S, Yoon J, Hwang JI, Suh PG, Sheng M, Kim E: Characterization of the Shank family of synaptic proteins. Multiple genes, alternative splicing, and differential expression in brain and development. J Biol Chem. 1999, 274: 29510-29518. 10.1074/jbc.274.41.29510.CrossRefPubMed

16.

Sugiyama Y, Kawabata I, Sobue K, Okabe S: Determination of absolute protein numbers in single synapses by a GFP-based calibration technique. Nat Methods. 2005, 2: 677-684. 10.1038/nmeth783.CrossRefPubMed

17.

Sala C, Piech V, Wilson NR, Passafaro M, Liu G, Sheng M: Regulation of dendritic spine morphology and synaptic function by Shank and Homer. Neuron. 2001, 31: 115-130. 10.1016/S0896-6273(01)00339-7.CrossRefPubMed

18.

Hung AY, Futai K, Sala C, Valtschanoff JG, Ryu J, Woodworth MA, Kidd FL, Sung CC, Miyakawa T, Bear MF: Smaller dendritic spines, weaker synaptic transmission, but enhanced spatial learning in mice lacking Shank1. J Neurosci. 2008, 28: 1697-1708. 10.1523/JNEUROSCI.3032-07.2008.PubMedCentralCrossRefPubMed

19.

Roussignol G, Ango F, Romorini S, Tu JC, Sala C, Worley PF, Bockaert J, Fagni L: Shank expression is sufficient to induce functional dendritic spine synapses in aspiny neurons. J Neurosci. 2005, 25: 3560-3570. 10.1523/JNEUROSCI.4354-04.2005.CrossRefPubMed

20.

Phelan MC, Rogers RC, Saul RA, Stapleton GA, Sweet K, McDermid H, Shaw SR, Claytor J, Willis J, Kelly DP: 22q13 deletion syndrome. Am J Med Genet. 2001, 101: 91-99. 10.1002/1096-8628(20010615)101:2<91::AID-AJMG1340>3.0.CO;2-C.CrossRefPubMed

21.

Luciani JJ, de Mas P, Depetris D, Mignon-Ravix C, Bottani A, Prieur M, Jonveaux P, Philippe A, Bourrouillou G, de Martinville B: Telomeric 22q13 deletions resulting from rings, simple deletions, and translocations: cytogenetic, molecular, and clinical analyses of 32 new observations. J Med Genet. 2003, 40: 690-696. 10.1136/jmg.40.9.690.PubMedCentralCrossRefPubMed

22.

Wilson HL, Wong AC, Shaw SR, Tse WY, Stapleton GA, Phelan MC, Hu S, Marshall J, McDermid HE: Molecular characterisation of the 22q13 deletion syndrome supports the role of haploinsufficiency of SHANK3/PROSAP2 in the major neurological symptoms. J Med Genet. 2003, 40: 575-584. 10.1136/jmg.40.8.575.PubMedCentralCrossRefPubMed

23.

Bonaglia MC, Giorda R, Borgatti R, Felisari G, Gagliardi C, Selicorni A, Zuffardi O: Disruption of the ProSAP2 gene in a t(12;22)(q24.1;q13.3) is associated with the 22q13.3 deletion syndrome. Am J Hum Genet. 2001, 69: 261-268. 10.1086/321293.PubMedCentralCrossRefPubMed

24.

Bonaglia MC, Giorda R, Mani E, Aceti G, Anderlid BM, Baroncini A, Pramparo T, Zuffardi O: Identification of a recurrent breakpoint within the SHANK3 gene in the 22q13.3 deletion syndrome. J Med Genet. 2006, 43: 822-828. 10.1136/jmg.2005.038604.PubMedCentralCrossRefPubMed

25.

Phelan MC: Deletion 22q13.3 syndrome. Orphanet Journal of Rare Diseases. 2008, 3: 14-10.1186/1750-1172-3-14.PubMedCentralCrossRefPubMed

26.

Durand CM, Betancur C, Boeckers TM, Bockmann J, Chaste P, Fauchereau F, Nygren G, Rastam M, Gillberg IC, Anckarsater H: Mutations in the gene encoding the synaptic scaffolding protein SHANK3 are associated with autism spectrum disorders. Nat Genet. 2007, 39: 25-27. 10.1038/ng1933.PubMedCentralCrossRefPubMed

27.

Moessner R, Marshall CR, Sutcliffe JS, Skaug J, Pinto D, Vincent J, Zwaigenbaum L, Fernandez B, Roberts W, Szatmari P, Scherer SW: Contribution of SHANK3 mutations to autism spectrum disorder. Am J Hum Genet. 2007, 81: 1289-1297. 10.1086/522590.PubMedCentralCrossRefPubMed

28.

Gauthier J, Spiegelman D, Piton A, Lafreniere RG, Laurent S, St-Onge J, Lapointe L, Hamdan FF, Cossette P, Mottron L: Novel de novo SHANK3 mutation in autistic patients. Am J Med Genet B Neuropsychiatr Genet. 2009, 150B: 421-424. 10.1002/ajmg.b.30822.CrossRefPubMed

29.

Gauthier J, Champagne N, Lafreniere RG, Xiong L, Spiegelman D, Brustein E, Lapointe M, Peng H, Cote M, Noreau A: De novo mutations in the gene encoding the synaptic scaffolding protein SHANK3 in patients ascertained for schizophrenia. Proc Natl Acad Sci USA. 2010, 107: 7863-7868. 10.1073/pnas.0906232107.PubMedCentralCrossRefPubMed

30.

Berkel S, Marshall CR, Weiss B, Howe J, Roeth R, Moog U, Endris V, Roberts W, Szatmari P, Pinto D: Mutations in the SHANK2 synaptic scaffolding gene in autism spectrum disorder and mental retardation. Nat Genet. 2010, 42: 489-491. 10.1038/ng.589.CrossRefPubMed

31.

Pinto D, Pagnamenta AT, Klei L, Anney R, Merico D, Regan R, Conroy J, Magalhaes TR, Correia C, Abrahams BS: Functional impact of global rare copy number variation in autism spectrum disorders. Nature. 2010, 466: 368-372. 10.1038/nature09146.PubMedCentralCrossRefPubMed

32.

Hughes ED, Qu YY, Genik SJ, Lyons RH, Pacheco CD, Lieberman AP, Samuelson LC, Nasonkin IO, Camper SA, Van Keuren ML, Saunders TL: Genetic variation in C57BL/6 ES cell lines and genetic instability in the Bruce4 C57BL/6 ES cell line. Mamm Genome. 2007, 18: 549-558. 10.1007/s00335-007-9054-0.CrossRefPubMed

33.

Hellemans J, Mortier G, De Paepe A, Speleman F, Vandesompele J: qBase relative quantification framework and software for management and automated analysis of real-time quantitative PCR data. Genome Biol. 2007, 8: R19-10.1186/gb-2007-8-2-r19.PubMedCentralCrossRefPubMed

34.

Yang Y, Wang XB, Frerking M, Zhou Q: Spine expansion and stabilization associated with long-term potentiation. J Neurosci. 2008, 28: 5740-5751. 10.1523/JNEUROSCI.3998-07.2008.PubMedCentralCrossRefPubMed

35.

Bozdagi O, Wang XB, Nikitczuk JS, Anderson TR, Bloss EB, Radice GL, Zhou Q, Benson DL, Huntley GW: Persistence of coordinated long-term potentiation and dendritic spine enlargement at mature hippocampal CA1 synapses requires N-cadherin. J Neurosci. 2010, 30: 9984-9989. 10.1523/JNEUROSCI.1223-10.2010.PubMedCentralCrossRefPubMed

36.

Holtmaat AJ, Trachtenberg JT, Wilbrecht L, Shepherd GM, Zhang X, Knott GW, Svoboda K: Transient and persistent dendritic spines in the neocortex in vivo. Neuron. 2005, 45: 279-291. 10.1016/j.neuron.2005.01.003.CrossRefPubMed

37.

Wearne SL, Rodriguez A, Ehlenberger DB, Rocher AB, Henderson SC, Hof PR: New techniques for imaging, digitization and analysis of three-dimensional neural morphology on multiple scales. Neuroscience. 2005, 136: 661-680. 10.1016/j.neuroscience.2005.05.053.CrossRefPubMed

38.

Rodriguez A, Ehlenberger DB, Dickstein DL, Hof PR, Wearne SL: Automated three-dimensional detection and shape classification of dendritic spines from fluorescence microscopy images. PLoS One. 2008, 3: e1997-10.1371/journal.pone.0001997.PubMedCentralCrossRefPubMed

39.

Rodriguez A, Ehlenberger DB, Hof PR, Wearne SL: Rayburst sampling, an algorithm for automated three-dimensional shape analysis from laser scanning microscopy images. Nat Protoc. 2006, 1: 2152-2161. 10.1038/nprot.2006.313.CrossRefPubMed

40.

Bozdagi O, Rich E, Tronel S, Sadahiro M, Patterson K, Shapiro ML, Alberini CM, Huntley GW, Salton SR: The neurotrophin-inducible gene Vgf regulates hippocampal function and behavior through a brain-derived neurotrophic factor-dependent mechanism. J Neurosci. 2008, 28: 9857-9869. 10.1523/JNEUROSCI.3145-08.2008.PubMedCentralCrossRefPubMed

41.

Huber KM, Kayser MS, Bear MF: Role for rapid dendritic protein synthesis in hippocampal mGluR-dependent long-term depression. Science. 2000, 288: 1254-1257. 10.1126/science.288.5469.1254.CrossRefPubMed

42.

Dumutriu D, Hara Y, Berger S, Wadsworth S, Morrison J: Volume assisted measurement of puncta in 2 dimensions (VAMP2D): A new tool for accurate size quantification in fluorescent imaging. Society for Neurosience Annual Meeting. 2009, Chicago, IL: Society for Neuroscience Abstracts

43.

Scattoni ML, Gandhy SU, Ricceri L, Crawley JN: Unusual repertoire of vocalizations in the BTBR T+tf/J mouse model of autism. PLoS One. 2008, 3: e3067-10.1371/journal.pone.0003067.PubMedCentralCrossRefPubMed

44.

Silverman JL, Turner SM, Barkan CL, Tolu SS, Saxena R, Hung AY, Sheng M, Crawley JN: Sociability and motor functions in Shank1 mutant mice. Brain Res. 2010,

45.

Scattoni ML, Ricceri L, Crawley JN: Unusual repertoire of vocalizations in adult BTBR T+tf/J mice during three types of social encounters. Genes Brain Behav. 2010

46.

Yang M, Crawley JN: Simple behavioral assessment of mouse olfaction. Curr Protoc Neurosci. 2009, Chapter 8 (Unit 8): 24-PubMed

47.

Silverman JL, Yang M, Lord C, Crawley JN: Behavioural phenotyping assays for mouse models of autism. Nat Rev Neurosci. 2010, 11: 490-502. 10.1038/nrn2851.PubMedCentralCrossRefPubMed

48.

Matsuzaki M, Honkura N, Ellis-Davies GC, Kasai H: Structural basis of long-term potentiation in single dendritic spines. Nature. 2004, 429: 761-766. 10.1038/nature02617.PubMedCentralCrossRefPubMed

49.

Kopec CD, Real E, Kessels HW, Malinow R: GluR1 links structural and functional plasticity at excitatory synapses. J Neurosci. 2007, 27: 13706-13718. 10.1523/JNEUROSCI.3503-07.2007.CrossRefPubMed

50.

Nyby J: Ultrasonic vocalizations during sex behavior of male house mice (Mus musculus): a description. Behav Neural Biol. 1983, 39: 128-134. 10.1016/S0163-1047(83)90722-7.CrossRefPubMed

51.

Maunakea AK, Nagarajan RP, Bilenky M, Ballinger TJ, D'Souza C, Fouse SD, Johnson BE, Hong C, Nielsen C, Zhao Y: Conserved role of intragenic DNA methylation in regulating alternative promoters. Nature. 2010, 466: 253-257. 10.1038/nature09165.PubMedCentralCrossRefPubMed

52.

Kolevzon A, Cai G, Soorya L, Takahashi N, Grodberg D, Kajiwara Y, Willner JP, Tryfon A, Buxbaum JD: Analysis of a purported SHANK3 mutation in a boy with autism: Clinical impact of rare variant research in neurodevelopmental disabilities. Brain Res. 2010,

53.

Uchino S, Wada H, Honda S, Nakamura Y, Ondo Y, Uchiyama T, Tsutsumi M, Suzuki E, Hirasawa T, Kohsaka S: Direct interaction of post-synaptic density-95/Dlg/ZO-1 domain-containing synaptic molecule Shank3 with GluR1 alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptor. J Neurochem. 2006, 97: 1203-1214. 10.1111/j.1471-4159.2006.03831.x.CrossRefPubMed

54.

Malinow R, Malenka RC: AMPA receptor trafficking and synaptic plasticity. Annu Rev Neurosci. 2002, 25: 103-126. 10.1146/annurev.neuro.25.112701.142758.CrossRefPubMed

55.

Shi S, Hayashi Y, Esteban JA, Malinow R: Subunit-specific rules governing AMPA receptor trafficking to synapses in hippocampal pyramidal neurons. Cell. 2001, 105: 331-343. 10.1016/S0092-8674(01)00321-X.CrossRefPubMed

56.

Park M, Penick EC, Edwards JG, Kauer JA, Ehlers MD: Recycling endosomes supply AMPA receptors for LTP. Science. 2004, 305: 1972-1975. 10.1126/science.1102026.CrossRefPubMed

57.

Ehrlich I, Malinow R: Postsynaptic density 95 controls AMPA receptor incorporation during long-term potentiation and experience-driven synaptic plasticity. J Neurosci. 2004, 24: 916-927. 10.1523/JNEUROSCI.4733-03.2004.CrossRefPubMed

58.

El-Husseini AE, Schnell E, Chetkovich DM, Nicoll RA, Bredt DS: PSD-95 involvement in maturation of excitatory synapses. Science. 2000, 290: 1364-1368.PubMed

59.

Stein V, House DR, Bredt DS, Nicoll RA: Postsynaptic density-95 mimics and occludes hippocampal long-term potentiation and enhances long-term depression. J Neurosci. 2003, 23: 5503-5506.PubMed

60.

King BH, Bostic JQ: An update on pharmacologic treatments for autism spectrum disorders. Child Adolesc Psychiatr Clin N Am. 2006, 15: 161-175. 10.1016/j.chc.2005.08.005.CrossRefPubMed

61.

Bear MF, Huber KM, Warren ST: The mGluR theory of fragile X mental retardation. Trends Neurosci. 2004, 27: 370-377. 10.1016/j.tins.2004.04.009.CrossRefPubMed

62.

Ehninger D, Silva AJ: Genetics and neuropsychiatric disorders: treatment during adulthood. Nat Med. 2009, 15: 849-850. 10.1038/nm0809-849.CrossRefPubMed

63.

Tropea D, Giacometti E, Wilson NR, Beard C, McCurry C, Fu DD, Flannery R, Jaenisch R, Sur M: Partial reversal of Rett Syndrome-like symptoms in MeCP2 mutant mice. Proc Natl Acad Sci USA. 2009, 106: 2029-2034. 10.1073/pnas.0812394106.PubMedCentralCrossRefPubMed

64.

Ehninger D, de Vries PJ, Silva AJ: From mTOR to cognition: molecular and cellular mechanisms of cognitive impairments in tuberous sclerosis. J Intellect Disabil Res. 2009, 53: 838-851. 10.1111/j.1365-2788.2009.01208.x.PubMedCentralCrossRefPubMed

Title: Haploinsufficiency of the autism-associated Shank3 gene leads to deficits in synaptic function, social interaction, and social communication
Authors: Ozlem Bozdagi
Takeshi Sakurai
Danae Papapetrou
Xiaobin Wang
Dara L Dickstein
Nagahide Takahashi
Yuji Kajiwara
Mu Yang
Adam M Katz
Maria Luisa Scattoni
Mark J Harris
Roheeni Saxena
Jill L Silverman
Jacqueline N Crawley
Qiang Zhou
Patrick R Hof
Joseph D Buxbaum
Publication date: 01-12-2010
Publisher: BioMed Central
Published in: Molecular Autism / Issue 1/2010
Electronic ISSN: 2040-2392
DOI: https://doi.org/10.1186/2040-2392-1-15

Keynote webinar | Spotlight on sleep in brain health

Springer Medicine

Haploinsufficiency of the autism-associated Shank3 gene leads to deficits in synaptic function, social interaction, and social communication

Abstract

Background

Methods

Results

Conclusions

Keynote webinar | Spotlight on sleep in brain health

Springer Medicine

Abstract

Background

Methods

Results

Conclusions

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