Regulatory roles of perineuronal nets and semaphorin 3A in the postnatal maturation of the central vestibular circuitry for graviceptive reflex

  • Chun-Wai Ma
  • Pui-Yi Kwan
  • Kenneth Lap-Kei Wu
  • Daisy Kwok-Yan Shum
  • Ying-Shing Chan
Original Article


Perineuronal nets (PN) restrict neuronal plasticity in the adult brain. We hypothesize that activity-dependent consolidation of PN is required for functional maturation of behavioral circuits. Using the postnatal maturation of brainstem vestibular nucleus (VN) circuits as a model system, we report a neonatal period in which consolidation of central vestibular circuitry for graviception is accompanied by activity-dependent consolidation of chondroitin sulfate (CS)-rich PN around GABAergic neurons in the VN. Postnatal onset of negative geotaxis was used as an indicator for functional maturation of vestibular circuits. Rats display negative geotaxis from postnatal day (P) 9, coinciding with the condensation of CS-rich PN around GABAergic interneurons in the VN. Delaying PN formation, by removal of primordial CS moieties on VN with chondroitinase ABC (ChABC) treatment at P6, postponed emergence of negative geotaxis to P13. Similar postponement was observed following inhibition of GABAergic transmission with bicuculline, in line with the reported role of PN in increasing excitability of parvalbumin neurons. We further reasoned that PN-CS restricts bioavailability of plasticity-inducing factors such as semaphorin 3A (Sema3A) to bring about circuit maturation. Treatment of VN explants with ChABC to liberate PN-bound Sema3A resulted in dendritic growth and arborization, implicating structural plasticity that delays synapse formation. Evidence is thus provided for the role of PN-CS–Sema3A in regulating structural and circuit plasticity at VN interneurons with impacts on the development of graviceptive postural control.


Perineuronal chondroitin sulfates Semaphorin 3A Vestibular system Development 



The authors are grateful to Simon S.M. Chan, Kimmy F.L. Tsang and Alice Y.Y. Lui of The University of Hong Kong for their excellent assistance in the experiments.


This work was supported by grants of the Hong Kong Research Grants Council (RGC HKU77681212, 17125115, N_HKU735/14).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standard of the institution of practice at which the studies were conducted. All animal protocols and procedures were performed in accordance to the Guide for the Care and Use of Laboratory Animals (NIH, 2011) and approved by The University of Hong Kong Committee on the Use of Live Animals in Teaching and Research. This article does not contain any human participants performed by any of the authors.


  1. Altman J, Bayer SA (1980) Development of the brain stem in the rat. III. Thymidine-radiographic study of the time of origin of neurons of the vestibular and auditory nuclei of the upper medulla. J Comp Neurol 194(4):877–904CrossRefGoogle Scholar
  2. Balmer TS (2016) Perineuronal nets enhance the excitability of fast-spiking neurons. eNeuro 3(4):0112–0116CrossRefGoogle Scholar
  3. Balmer TS, Carels VM, Frisch JL, Nick TA (2009) Modulation of perineuronal nets and parvalbumin with developmental song learning. J Neurosci 29(41):12878–12885. CrossRefPubMedPubMedCentralGoogle Scholar
  4. Bandtlow CE, Zimmermann DR (2000) Proteoglycans in the developing brain: new conceptual insights for old proteins. Physiol Rev 80(4):1267–1290CrossRefGoogle Scholar
  5. Beurdeley M, Spatazza J, Lee HH, Sugiyama S, Bernard C, Di Nardo AA, Hensch TK, Prochiantz A (2012) Otx2 binding to perineuronal nets persistently regulates plasticity in the mature visual cortex. J Neurosci 32(27):9429–9437CrossRefGoogle Scholar
  6. Bouet V, Wubbels RJ, de Jong HA, Gramsbergen A (2004) Behavioural consequences of hypergravity in developing rats. Brain Res Dev Brain Res 153(1):69–78. CrossRefPubMedGoogle Scholar
  7. Buttner U, Straube A, Kurzan R (1992) Oculomotor effects of gamma-aminobutyric acid agonists and antagonists in the vestibular nuclei of the alert monkey. Ann N Y Acad Sci 656:645–659CrossRefGoogle Scholar
  8. Carulli D, Rhodes KE, Brown DJ, Bonnert TP, Pollack SJ, Oliver K, Strata P, Fawcett JW (2006) Composition of perineuronal nets in the adult rat cerebellum and the cellular origin of their components. J Comp Neurol 494(4):559–577. CrossRefPubMedGoogle Scholar
  9. Carulli D, Pizzorusso T, Kwok JC, Putignano E, Poli A, Forostyak S, Andrews MR, Deepa SS, Glant TT, Fawcett JW (2010) Animals lacking link protein have attenuated perineuronal nets and persistent plasticity. Brain 133(8):2331–2347CrossRefGoogle Scholar
  10. Carulli D, Foscarin S, Faralli A, Pajaj E, Rossi F (2013) Modulation of semaphorin3A in perineuronal nets during structural plasticity in the adult cerebellum. Mol Cell Neurosci 57:10–22. CrossRefPubMedGoogle Scholar
  11. Chirumamilla S, Sun D, Bullock M, Colello R (2002) Traumatic brain injury induced cell proliferation in the adult mammalian central nervous system. J Neurotrauma 19(6):693–703CrossRefGoogle Scholar
  12. Crozier WJ, Pincus G (1926) Tropisms of Mammals. Proc Natl Acad Sci USA 12(10):612–616CrossRefGoogle Scholar
  13. Cullheim S, Thams S (2007) The microglial networks of the brain and their role in neuronal network plasticity after lesion. Brain Res Rev 55(1):89–96CrossRefGoogle Scholar
  14. Curthoys I (1982) Postnatal developmental changes in the response of rat primary horizontal semicircular canal neurons to sinusoidal angular accelerations. Exp Brain Res 47(2):295–300PubMedGoogle Scholar
  15. Deliagina TG, Orlovsky GN (2002) Comparative neurobiology of postural control. Curr Opin Neurobiol 12(6):652–657CrossRefGoogle Scholar
  16. Dick G, Tan CL, Alves JN, Ehlert EM, Miller GM, Hsieh-Wilson LC, Sugahara K, Oosterhof A, van Kuppevelt TH, Verhaagen J (2013) Semaphorin 3A binds to the perineuronal nets via chondroitin sulfate type E motifs in rodent brains. J Biol Chem 288(38):27384–27395CrossRefGoogle Scholar
  17. Dieterich M, Straube A, Brandt T, Paulus W, Buttner U (1991) The effects of baclofen and cholinergic drugs on upbeat and downbeat nystagmus. J Neurol Neurosurg Psychiatry 54(7):627–632CrossRefGoogle Scholar
  18. Dow RS (1938) The effects of unilateral and bilateral labyrinthectomy in monkey, baboon and chimpanzee. Am J Physiol Legacy Content 121(2):392–399CrossRefGoogle Scholar
  19. Geissler M, Gottschling C, Aguado A, Rauch U, Wetzel CH, Hatt H, Faissner A (2013) Primary hippocampal neurons, which lack four crucial extracellular matrix molecules, display abnormalities of synaptic structure and function and severe deficits in perineuronal net formation. J Neurosci 33(18):7742–7755. CrossRefPubMedGoogle Scholar
  20. Gliddon CM, Darlington CL, Smith PF (2005) GABAergic systems in the vestibular nucleus and their contribution to vestibular compensation. Prog Neurobiol 75(1):53–81. CrossRefPubMedGoogle Scholar
  21. Glykys J, Dzhala V, Egawa K, Balena T, Saponjian Y, Kuchibhotla KV, Bacskai BJ, Kahle KT, Zeuthen T, Staley KJ (2014) Local impermeant anions establish the neuronal chloride concentration. Science 343(6171):670–675. CrossRefPubMedPubMedCentralGoogle Scholar
  22. Hartig W, Brauer K, Bigl V, Bruckner G (1994) Chondroitin sulfate proteoglycan-immunoreactivity of lectin-labeled perineuronal nets around parvalbumin-containing neurons. Brain Res 635(1–2):307–311CrossRefGoogle Scholar
  23. Hensch TK (2005) Critical period plasticity in local cortical circuits. Nat Rev Neurosci 6(11):877–888. CrossRefPubMedGoogle Scholar
  24. Janušonis S, Fite KV (2001) Diurnal variation of c-Fos expression in subdivisions of the dorsal raphe nucleus of the Mongolian gerbil (Meriones unguiculatus). J Comp Neurol 440(1):31–42CrossRefGoogle Scholar
  25. Jones LL, Margolis RU, Tuszynski MH (2003) The chondroitin sulfate proteoglycans neurocan, brevican, phosphacan, and versican are differentially regulated following spinal cord injury. Exp Neurol 182(2):399–411CrossRefGoogle Scholar
  26. Kamikouchi A, Inagaki HK, Effertz T, Hendrich O, Fiala A, Gopfert MC, Ito K (2009) The neural basis of Drosophila gravity-sensing and hearing. Nature 458(7235):165–171. CrossRefPubMedGoogle Scholar
  27. Kato R, Iwamoto Y, Yoshida K (2003) Contribution of GABAergic inhibition to the responses of secondary vestibular neurons to head rotation in the rat. Neurosci Res 46(4):499–508CrossRefGoogle Scholar
  28. Lai CH, Chan YS (2001) Spontaneous discharge and response characteristics of central otolith neurons of rats during postnatal development. Neuroscience 103(1):275–288CrossRefGoogle Scholar
  29. Lai CH, Tse YC, Shum DK, Yung KK, Chan YS (2004) Fos expression in otolith-related brainstem neurons of postnatal rats following off-vertical axis rotation. J Comp Neurol 470(3):282–296. CrossRefPubMedGoogle Scholar
  30. Lai SK, Lai CH, Yung KK, Shum DK, Chan YS (2006) Maturation of otolith-related brainstem neurons in the detection of vertical linear acceleration in rats. Eur J Neurosci 23(9):2431–2446. CrossRefPubMedGoogle Scholar
  31. Lai CH, Ma CW, Lai SK, Han L, Wong HM, Yeung KW, Shum DK, Chan YS (2016) Maturation of glutamatergic transmission in the vestibulo-olivary pathway impacts on the registration of head rotational signals in the brainstem of rats. Brain Struct Funct 221(1):217–238. CrossRefPubMedGoogle Scholar
  32. Liu J, Chau CH, Liu H, Jang BR, Li X, Chan YS, Shum DK (2006) Upregulation of chondroitin 6-sulphotransferase-1 facilitates Schwann cell migration during axonal growth. J Cell Sci 119(Pt 5):933–942. CrossRefPubMedGoogle Scholar
  33. Ma CW, Zhang FX, Lai CH, Lai SK, Yung KK, Shum DK, Chan YS (2013) Postnatal expression of TrkB receptor in rat vestibular nuclear neurons responsive to horizontal and vertical linear accelerations. J Comp Neurol 521(3):612–625. CrossRefPubMedGoogle Scholar
  34. Marlinsky V (1992) Activity of lateral vestibular nucleus neurons during locomotion in the decerebrate guinea pig. Exp Brain Res 90(3):583–588CrossRefGoogle Scholar
  35. Matsuyama K, Drew T (2000) Vestibulospinal and reticulospinal neuronal activity during locomotion in the intact cat. I. Walking on a level surface. J Neurophysiol 84(5):2237–2256. CrossRefPubMedGoogle Scholar
  36. McRae PA, Rocco MM, Kelly G, Brumberg JC, Matthews RT (2007) Sensory deprivation alters aggrecan and perineuronal net expression in the mouse barrel cortex. J Neurosci 27(20):5405–5413. CrossRefPubMedGoogle Scholar
  37. Miyata S, Komatsu Y, Yoshimura Y, Taya C, Kitagawa H (2012) Persistent cortical plasticity by upregulation of chondroitin 6-sulfation. Nat Neurosci 15(3):414–422. (S411–412) CrossRefPubMedGoogle Scholar
  38. Moratalla R, Elibol B, Vallejo M, Graybiel AM (1996) Network-level changes in expression of inducible Fos–Jun proteins in the striatum during chronic cocaine treatment and withdrawal. Neuron 17(1):147–156CrossRefGoogle Scholar
  39. Morita A, Yamashita N, Sasaki Y, Uchida Y, Nakajima O, Nakamura F, Yagi T, Taniguchi M, Usui H, Katoh-Semba R, Takei K, Goshima Y (2006) Regulation of dendritic branching and spine maturation by semaphorin3A-Fyn signaling. J Neurosci 26(11):2971–2980. CrossRefPubMedGoogle Scholar
  40. Murphy GJ, Du Lac S (2001) Postnatal development of spike generation in rat medial vestibular nucleus neurons. J Neurophysiol 85(5):1899–1906CrossRefGoogle Scholar
  41. Nabel EM, Morishita H (2013) Regulating critical period plasticity: insight from the visual system to fear circuitry for therapeutic interventions. Front Psychiatry 4:146. CrossRefPubMedPubMedCentralGoogle Scholar
  42. Nakamura J, Sasa M, Takaori S (1989) Ethanol potentiates the effect of gamma-aminobutyric acid on medial vestibular nucleus neurons responding to horizontal rotation. Life Sci 45(11):971–978CrossRefGoogle Scholar
  43. Orlando C, Ster J, Gerber U, Fawcett JW, Raineteau O (2012) Perisynaptic chondroitin sulfate proteoglycans restrict structural plasticity in an integrin-dependent manner. J Neurosci 32(50):18009–18017. CrossRefPubMedGoogle Scholar
  44. Pincus G, Crozier WJ (1929) On the geotropic response in young rats. Proc Natl Acad Sci USA 15(7):581–586CrossRefGoogle Scholar
  45. Pizzorusso T, Medini P, Berardi N, Chierzi S, Fawcett JW, Maffei L (2002) Reactivation of ocular dominance plasticity in the adult visual cortex. Science 298(5596):1248–1251. CrossRefPubMedGoogle Scholar
  46. Pizzorusso T, Medini P, Landi S, Baldini S, Berardi N, Maffei L (2006) Structural and functional recovery from early monocular deprivation in adult rats. Proc Natl Acad Sci USA 103(22):8517–8522. CrossRefPubMedGoogle Scholar
  47. Reber A, Poitevin B, Leroy MH, Nzobounsana V (1996) Optokinetic and vestibulo-ocular reflex adjustment by GABA antagonists. Behav Brain Res 81(1–2):89–97CrossRefGoogle Scholar
  48. Schnupp JW, King AJ, Smith AL, Thompson ID (1995) NMDA-receptor antagonists disrupt the formation of the auditory space map in the mammalian superior colliculus. J Neurosci 15(2):1516–1531CrossRefGoogle Scholar
  49. Shelly M, Cancedda L, Lim BK, Popescu AT, Cheng PL, Gao H, Poo MM (2011) Semaphorin3A regulates neuronal polarization by suppressing axon formation and promoting dendrite growth. Neuron 71(3):433–446. CrossRefPubMedPubMedCentralGoogle Scholar
  50. Smith AL, Cordery PM, Thompson ID (1995) Manufacture and release characteristics of Elvax polymers containing glutamate receptor antagonists. J Neurosci Methods 60(1–2):211–217CrossRefGoogle Scholar
  51. Tighilet B, Lacour M (2001) Gamma amino butyric acid (GABA) immunoreactivity in the vestibular nuclei of normal and unilateral vestibular neurectomized cats. Eur J Neurosci 13(12):2255–2267CrossRefGoogle Scholar
  52. Uesaka N, Uchigashima M, Mikuni T, Nakazawa T, Nakao H, Hirai H, Aiba A, Watanabe M, Kano M (2014) Retrograde semaphorin signaling regulates synapse elimination in the developing mouse brain. Science 344(6187):1020–1023. CrossRefPubMedGoogle Scholar
  53. Wills TJ, Muessig L, Cacucci F (2014) The development of spatial behaviour and the hippocampal neural representation of space. Phil Trans R Soc B 369(1635):20130409CrossRefGoogle Scholar
  54. Wong HM, Yeung KW, Lam KO, Tam V, Chu PK, Luk KD, Cheung KM (2010) A biodegradable polymer-based coating to control the performance of magnesium alloy orthopaedic implants. Biomaterials 31(8):2084–2096. CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.School of Biomedical Sciences, Li Ka Shing Faculty of MedicineThe University of Hong KongHong Kong SARPeople’s Republic of China
  2. 2.State Key Laboratory of Brain and Cognitive Sciences, Li Ka Shing Faculty of MedicineThe University of Hong KongHong Kong SARPeople’s Republic of China

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