Behavioral decline in Shank3 ^Δex4-22 mice during early adulthood parallels cerebellar granule cell glutamatergic synaptic changes.

Publisher: BioMed Central Country of Publication: England NLM ID: 101534222 Publication Model: Electronic Cited Medium: Internet ISSN: 2040-2392 (Electronic) NLM ISO Abbreviation: Mol Autism Subsets: MEDLINE

Original Publication: [London] : BioMed Central

Nerve Tissue Proteins*/metabolism ; Nerve Tissue Proteins*/genetics ; Mice, Knockout* ; Synapses*/metabolism ; Behavior, Animal* ; Excitatory Postsynaptic Potentials* ; Cerebellum*/metabolism; Animals ; Male ; Mice ; Female ; Microfilament Proteins/metabolism ; Microfilament Proteins/genetics ; Glutamic Acid/metabolism ; Exons ; Mice, Inbred C57BL ; Neurons/metabolism

Background: SHANK3, a gene encoding a synaptic scaffolding protein, is implicated in autism spectrum disorder (ASD) and is disrupted in Phelan-McDermid syndrome (PMS). Despite evidence of regression or worsening of ASD-like symptoms in individuals with PMS, the underlying mechanisms remain unclear. Although Shank3 is highly expressed in the cerebellar cortical granule cells, its role in cerebellar function and contribution to behavioral deficits in ASD models are unknown. This study investigates behavioral changes and cerebellar synaptic alterations in Shank3 ^Δex4-22 mice at two developmental stages.
Methods: Shank3 ^Δex4-22 wildtype, heterozygous, and homozygous knockout mice lacking exons 4-22 (all functional isoforms) were subjected to a behavioral battery in both juvenile (5-7 weeks old) and adult (3-5 months old) mouse cohorts of both sexes. Immunostaining was used to show the expression of Shank3 in the cerebellar cortex. Spontaneous excitatory postsynaptic currents (sEPSCs) from cerebellar granule cells (CGCs) were recorded by whole-cell patch-clamp electrophysiology.
Results: Deletion of Shank3 caused deficits in motor function, heightened anxiety, and repetitive behaviors. These genotype-dependent behavioral alterations were more prominent in adult mice than in juveniles. Reduced social preference was only identified in adult Shank3 ^Δex4-22 knockout male mice, while self-grooming was uniquely elevated in males across both age groups. Heterozygous mice showed little to no changes in behavioral phenotypes in most behavioral tests. Immunofluorescence staining indicated the presence of Shank3 predominantly in the dendrite-containing rosette-like structures in CGCs, colocalizing with presynaptic markers of glutamatergic mossy fiber. Electrophysiological findings identified a parallel relationship between the age-related exacerbation of behavioral impairments and the enhancement of sEPSC amplitude in CGCs.
Limitations: Other behavioral tests of muscle strength (grip strength test), memory (Barnes/water maze), and communication (ultrasonic vocalization), were not performed. Further study is necessary to elucidate how Shank3 modulates synaptic function at the mossy fiber-granule cell synapse in the cerebellum and whether these changes shape the behavioral phenotype.
Conclusions: Our findings reveal an age-related exacerbation of behavioral impairments in Shank3 ^Δex4-22 mutant mice. These results suggest that Shank3 may alter the function of glutamatergic receptors at the mossy fiber-cerebellar granule cell synapse as a potential mechanism causing cerebellar disruption in ASD.
Competing Interests: Declarations. Ethics approval: All procedures involving animals were performed in accordance with protocols approved by the Institutional Animal Care and Use Committee at Southern Illinois Universe – School of Medicine or the University of Idaho. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests.
(© 2024. The Author(s).)

Update of: Res Sq. 2024 Sep 06:rs.3.rs-4888950. doi: 10.21203/rs.3.rs-4888950/v1. (PMID: 39281868)

Front Behav Neurosci. 2023 Jan 09;16:1051175. (PMID: 36699652)
Mol Autism. 2016 Oct 21;7:44. (PMID: 27790361)
Nat Genet. 2007 Jan;39(1):25-7. (PMID: 17173049)
J Microsc. 1993 Mar;169(3):375-382. (PMID: 33930978)
Mol Autism. 2018 Apr 27;9:31. (PMID: 29719671)
Synapse. 2011 Nov;65(11):1204-12. (PMID: 21638338)
Front Neurosci. 2015 Nov 06;9:420. (PMID: 26594141)
Elife. 2020 Jul 08;9:. (PMID: 32639229)
PLoS Genet. 2014 Sep 04;10(9):e1004580. (PMID: 25188300)
Autism Res. 2016 Mar;9(3):350-75. (PMID: 26559786)
Nat Protoc. 2020 Oct;15(10):3464-3477. (PMID: 32895524)
Elife. 2021 Sep 20;10:. (PMID: 34542410)
Am J Med Genet. 1992 Jul 15;43(5):872-6. (PMID: 1353666)
Neuroimage Clin. 2015 Feb 20;7:631-9. (PMID: 25844317)
PLoS Comput Biol. 2013;9(7):e1003128. (PMID: 23935468)
Cerebellum. 2017 Feb;16(1):203-229. (PMID: 26873754)
Front Mol Neurosci. 2023 Mar 16;16:1139118. (PMID: 37008785)
Psychopharmacology (Berl). 2009 Jun;204(2):361-73. (PMID: 19189082)
J Biol Chem. 2001 Oct 26;276(43):40104-12. (PMID: 11509555)
Cerebellum. 2016 Dec;15(6):732-743. (PMID: 26585120)
Am J Hum Genet. 2014 May 1;94(5):677-94. (PMID: 24768552)
J Biol Chem. 1999 Oct 8;274(41):29510-8. (PMID: 10506216)
Eur J Neurosci. 2023 Jun;57(12):1966-1979. (PMID: 37165567)
Sci Rep. 2017 Sep 20;7(1):12008. (PMID: 28931838)
Cerebellum. 2012 Sep;11(3):777-807. (PMID: 22370873)
Front Neurol. 2017 Nov 27;8:615. (PMID: 29230189)
Front Syst Neurosci. 2014 May 20;8:92. (PMID: 24904314)
PLoS One. 2014 Feb 24;9(2):e81255. (PMID: 24586223)
Front Neurol. 2016 Aug 10;7:124. (PMID: 27559329)
Nat Rev Neurosci. 2017 Mar;18(3):147-157. (PMID: 28179641)
Semin Fetal Neonatal Med. 2016 Oct;21(5):349-55. (PMID: 27179922)
Autism Res. 2017 Feb;10(2):251-266. (PMID: 27220548)
J Neuropsychiatry Clin Neurosci. 2004 Summer;16(3):367-78. (PMID: 15377747)
NPJ Genom Med. 2019 Aug 23;4:19. (PMID: 31452935)
Hum Mol Genet. 2015 Oct 15;24(R1):R24-31. (PMID: 26188008)
Mol Autism. 2014 Apr 25;5:30. (PMID: 25071925)
Nature. 2011 Apr 28;472(7344):437-42. (PMID: 21423165)
Dev Med Child Neurol. 2023 Jul;65(7):917-925. (PMID: 36477723)
Methods Mol Biol. 2019;1916:105-111. (PMID: 30535688)
Mol Syndromol. 2012 Apr;2(3-5):186-201. (PMID: 22670140)
Int Rev Neurobiol. 1997;41:433-40. (PMID: 9378601)
Science. 1993 Apr 2;260(5104):95-7. (PMID: 8096653)
Int Rev Neurobiol. 2013;113:1-34. (PMID: 24290381)
Autism. 2018 Feb;22(2):195-204. (PMID: 29490485)
Mol Psychiatry. 2017 May;22(5):689-702. (PMID: 27021819)
Neuroimage. 2012 Jan 16;59(2):1560-70. (PMID: 21907811)
J Physiol. 2009 Dec 15;587(Pt 24):5843-57. (PMID: 19858226)
Philos Trans R Soc Lond B Biol Sci. 2003 Apr 29;358(1432):829-42. (PMID: 12740130)
J Pharmacol Toxicol Methods. 2020 Mar - Apr;102:106676. (PMID: 31954839)
Nat Rev Neurosci. 2011 Jun;12(6):327-44. (PMID: 21544091)
J Neurophysiol. 2017 Jul 1;118(1):267-279. (PMID: 28381493)
PLoS One. 2019 Mar 15;14(3):e0213921. (PMID: 30875393)
Eur J Med Genet. 2023 Jul;66(7):104771. (PMID: 37120079)
Brain. 2007 Oct;130(Pt 10):2646-60. (PMID: 17872929)
Nat Neurosci. 2015 May;18(5):718-27. (PMID: 25821914)
J Autism Dev Disord. 1995 Feb;25(1):1-18. (PMID: 7608030)
Nature. 2016 Feb 25;530(7591):481-4. (PMID: 26886798)
Front Neurosci. 2015 Sep 01;9:296. (PMID: 26388713)
J Neurodev Disord. 2016 Apr 26;8:16. (PMID: 27118998)
Acta Neuropathol. 2017 Oct;134(4):537-566. (PMID: 28584888)
J Neurosci. 2016 Aug 31;36(35):9019-25. (PMID: 27581446)
Trends Cell Biol. 2011 Oct;21(10):594-603. (PMID: 21840719)
Acta Neuropathol. 2010 Jun;119(6):755-70. (PMID: 20198484)
Cell Rep. 2015 Jun 9;11(9):1400-1413. (PMID: 26027926)
Transl Psychiatry. 2018 Apr 27;8(1):94. (PMID: 29700290)
Front Neuroanat. 2021 Dec 16;15:759948. (PMID: 34975418)
Elife. 2015 Jul 09;4:e06085. (PMID: 26158416)
Brain Behav Immun. 2023 Nov;114:311-324. (PMID: 37657643)
Mol Autism. 2013 Jun 11;4(1):17. (PMID: 23758743)
Elife. 2021 Jul 05;10:. (PMID: 34219651)
Neuron. 2014 Aug 6;83(3):518-32. (PMID: 25102558)
Front Mol Neurosci. 2013 Aug 05;6:19. (PMID: 23935565)
Brain Struct Funct. 2012 Apr;217(2):165-80. (PMID: 21814870)
Brain. 2009 Sep;132(Pt 9):2413-25. (PMID: 19389870)
MMWR Surveill Summ. 2020 Mar 27;69(4):1-12. (PMID: 32214087)
Neuron. 1999 Jul;23(3):569-82. (PMID: 10433268)
J Neurosci. 2002 Nov 15;22(22):9687-97. (PMID: 12427824)
Neuron. 2016 Jan 6;89(1):147-62. (PMID: 26687841)
Neurosci Bull. 2017 Apr;33(2):205-218. (PMID: 28271437)
J Neurodev Disord. 2014;6(1):39. (PMID: 25784960)
Neural Netw. 2008 Oct;21(8):1056-69. (PMID: 18603407)
J Neurosci. 2003 Jul 9;23(14):6074-85. (PMID: 12853426)
Mol Autism. 2010 Dec 17;1(1):15. (PMID: 21167025)
J Neurosci. 2013 Nov 20;33(47):18448-68. (PMID: 24259569)
Am J Psychiatry. 2014 Oct;171(10):1107-16. (PMID: 24873905)
Nat Rev Neurosci. 2019 May;20(5):298-313. (PMID: 30923348)
eNeuro. 2018 Oct 5;5(3):. (PMID: 30302388)
Dis Model Mech. 2014 Jun;7(6):667-81. (PMID: 24652766)
Mol Autism. 2019 Dec 24;10:50. (PMID: 31879555)
Hum Brain Mapp. 2018 Aug;39(8):3227-3240. (PMID: 29617056)
Cell Rep. 2021 Sep 21;36(12):109721. (PMID: 34551311)
J Neurosci. 2012 May 9;32(19):6525-41. (PMID: 22573675)
J Neurocytol. 1993 Sep;22(9):689-95. (PMID: 7903688)
Neurology. 2001 Jul 24;57(2):245-54. (PMID: 11468308)
Eur J Neurosci. 2023 Feb;57(4):607-618. (PMID: 36656446)
Pediatrics. 2007 Sep;120(3):584-93. (PMID: 17766532)
Front Neurosci. 2015 Nov 05;9:408. (PMID: 26594140)
J Neurodev Disord. 2020 Feb 12;12(1):7. (PMID: 32050889)
Trends Neurosci. 2009 Jan;32(1):30-40. (PMID: 18977038)
Neurotherapeutics. 2015 Jul;12(3):620-30. (PMID: 25894671)
J Psychiatr Res. 2017 Aug;91:139-144. (PMID: 28346892)
Nat Rev Neurosci. 2009 Sep;10(9):670-81. (PMID: 19693030)
Neurochem Res. 2011 Jul;36(7):1241-52. (PMID: 21243430)
Cell. 2020 Feb 6;180(3):568-584.e23. (PMID: 31981491)
Annu Rev Neurosci. 2009;32:413-34. (PMID: 19555291)
Front Cell Neurosci. 2015 Mar 19;9:94. (PMID: 25852484)
J Vis Exp. 2019 Jul 18;(149):. (PMID: 31380846)
Autism Res. 2017 Jan;10(1):42-65. (PMID: 27492494)
Curr Protoc Mouse Biol. 2012 Mar 01;2(1):37-53. (PMID: 26069004)
Eur J Neurosci. 1996 Jun;8(6):1182-9. (PMID: 8752588)
Mol Autism. 2013 Jun 11;4(1):18. (PMID: 23758760)
Nat Commun. 2016 May 10;7:11459. (PMID: 27161151)

P20 GM103408 United States GM NIGMS NIH HHS; R01 MH129749 United States MH NIMH NIH HHS; R01MH129749 United States NH NIH HHS; P20GM103408 United States NH NIH HHS

Keywords: AMPAR; Autism spectrum disorder; Cerebellum; Glutamate receptor; Granule cell; Mouse behavior; Phelan-McDermid syndrome; Shank3

0 (Shank3 protein, mouse)
0 (Nerve Tissue Proteins)
0 (Microfilament Proteins)
3KX376GY7L (Glutamic Acid)

Date Created: 20241205 Date Completed: 20241205 Latest Revision: 20241210

20241210

PMC11616285

10.1186/s13229-024-00628-y

39633421