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Vitronectin is Involved in the Morphological Transition of Neurites in Retinoic Acid-Induced Neurogenesis of Neuroblastoma Cell Line Neuro2a

  • Miyaka Sugahara
  • Yuri Nakaoki
  • Ayano Yamaguchi
  • Kei Hashimoto
  • Yasunori MiyamotoEmail author
Original Paper
  • 41 Downloads

Abstract

Vitronectin (Vtn), one of the extracellular matrix proteins, has been reported to result in cell cycle exit, neurite formation, and polarization of neural progenitor cells during neurogenesis. The underlying mechanism, however, has not been fully understood. In this study, we investigated the roles of Vtn and its integrin receptors, during the transition of neurites from multipolar to bipolar morphology, accompanying the cell cycle exit in neural progenitor cells. We used mouse neuroblastoma cell line Neuro2a as a model of neural progenitor cells which can induce cell cycle exit and the morphological transition of neurites by retinoic acid (RA)-stimulation. Treatment with an antibody for Vtn suppressed the RA-induced cell cycle exit and multipolar-to-bipolar transition. Furthermore, immunostaining results showed that in the cells displaying multipolar morphology Vtn was partially localized at the tips of neurites and in cells displaying bipolar morphology at both tips. This Vtn localization and multipolar-to-bipolar transition was perturbed by the transfection of a dominant negative mutant of cell polarity regulator Par6. In addition, a knockdown of β5 integrin, which is a receptor candidate for Vtn, affected the multipolar-to-bipolar transition. Taken together, these results suggest that Vtn regulates the multipolar-to-bipolar morphological transition via αvβ5 integrin.

Keywords

Vitronectin Integrin Morphological transition Cell cycle 

Abbreviations

Vtn

Vitronectin

RA

Retinoic acid

CGCP

Cerebellar granule cell precursor

DAPI

4′,6-Diamidino-2-phenylindole, dihydrochloride

BrdU

5-Bromodeoxyuridine

siRNA

Short-interfering RNA

Notes

Acknowledgements

We thank professor S. P. Atamas (University of Maryland School of Medicine, Baltimore, MD) for providing the integrin expression plasmids. We also thank T. Kobayashi (Ochanomizu University, Tokyo, Japan) for helpful comments. This work was funded by the Sasakawa Scientific Research Grant from The Japan Science Society and the Japan Society for the Promotion of Science (JSPS) KAKENHI Grant Number 17K07105 from the Ministry of Education, Culture, Sports, Science, and Technology of Japan.

Compliance with Ethical Standards

Conflict of interest

The authors declare no conflict of interest.

Ethical Approval

This article does not contain any studies with human participants or animals performed by any of the authors.

References

  1. 1.
    Miyata T, Ono Y, Okamoto M, Masaoka M, Sakakibara A, Kawaguchi A, Hashimoto M, Ogawa M (2010) Migration, early axonogenesis, and Reelin-dependent layer-forming behavior of early/posterior-born Purkinje cells in the developing mouse lateral cerebellum. Neural Dev 5:23CrossRefGoogle Scholar
  2. 2.
    Lui JH, Hansen DV, Kriegstein AR (2011) Development and evolution of the human neocortex. Cell 146:18–36CrossRefGoogle Scholar
  3. 3.
    Tabata H, Nakajima K (2003) Multipolar migration: the third mode of radial neuronal migration in the developing cerebral cortex. J Neurosci 23:9996–10001CrossRefGoogle Scholar
  4. 4.
    Kawauchi T, Chihama K, Nabeshima Y, Hoshino M (2006) Cdk5 phosphorylates and stabilizes p27kip1 contributing to actin organization and cortical neuronal migration. Nat Cell Biol 8:17–26CrossRefGoogle Scholar
  5. 5.
    Nakashima K, Umeshima H, Kengaku M (2015) Cerebellar granule cells are predominantly generated by terminal symmetric divisions of granule cell precursors. Dev Dyn 244:748–758CrossRefGoogle Scholar
  6. 6.
    Seiffert D, Iruela-Arispe ML, Sage EH, Loskutoff DJ (1995) Distribution of vitronectin mRNA during murine development. Dev Dyn 203:71–79CrossRefGoogle Scholar
  7. 7.
    Hayman EG, Pierschbacher MD, Ohgren Y, Ruoslahti E (1983) Serum spreading factor (vitronectin) is present at the cell surface and in tissues. Proc Natl Acad Sci USA 80:4003–4007CrossRefGoogle Scholar
  8. 8.
    Martinez-Morales JR, Barbas JA, Marti E, Bovolenta P, Edgar D, Rodriguez-Tebar A (1997) Vitronectin is expressed in the ventral region of the neural tube and promotes the differentiation of motor neurons. Development 124:5139–5147Google Scholar
  9. 9.
    Pons S, Marti E (2000) Sonic hedgehog synergizes with the extracellular matrix protein vitronectin to induce spinal motor neuron differentiation. Development 127:333–342Google Scholar
  10. 10.
    Martinez-Morales JR, Marti E, Frade JM, Rodriguez-Tebar A (1995) Developmentally regulated vitronectin influences cell differentiation, neuron survival and process outgrowth in the developing chicken retina. Neuroscience 68:245–253CrossRefGoogle Scholar
  11. 11.
    Grabham PW, Gallimore PH, Grand RJ (1992) Vitronectin is the major serum protein essential for NGF-mediated neurite outgrowth from PC12 cells. Exp Cell Res 202:337–344CrossRefGoogle Scholar
  12. 12.
    Pons S, Trejo JL, Martinez-Morales JR, Marti E (2001) Vitronectin regulates Sonic hedgehog activity during cerebellum development through CREB phosphorylation. Development 128:1481–1492Google Scholar
  13. 13.
    Murase S, Hayashi Y (1998) Concomitant expression of genes encoding integrin alpha v beta 5 heterodimer and vitronectin in growing parallel fibers of postnatal rat cerebellum: a possible role as mediators of parallel fiber elongation. J Comp Neurol 397:199–212CrossRefGoogle Scholar
  14. 14.
    Gupta SK, Meiri KF, Mahfooz K, Bharti U, Mani S (2010) Coordination between extrinsic extracellular matrix cues and intrinsic responses to orient the centrosome in polarizing cerebellar granule neurons. J Neurosci 30:2755–2766CrossRefGoogle Scholar
  15. 15.
    Abe A, Hashimoto K, Akiyama A, Iida M, Ikeda N, Hamano A, Watanabe R, Hayashi Y, Miyamoto Y (2018) alphavbeta5 integrin mediates the effect of vitronectin on the initial stage of differentiation in mouse cerebellar granule cell precursors. Brain Res 1691:94–104CrossRefGoogle Scholar
  16. 16.
    Hashimoto K, Sakane F, Ikeda N, Akiyama A, Sugahara M, Miyamoto Y (2016) Vitronectin promotes the progress of the initial differentiation stage in cerebellar granule cells. Mol Cell Neurosci 70:76–85CrossRefGoogle Scholar
  17. 17.
    Marzinke MA, Clagett-Dame M (2012) The all-trans retinoic acid (atRA)-regulated gene Calmin (Clmn) regulates cell cycle exit and neurite outgrowth in murine neuroblastoma (Neuro2a) cells. Exp Cell Res 318:85–93CrossRefGoogle Scholar
  18. 18.
    Mori Y, Matsui T, Omote D, Fukuda M (2013) Small GTPase Rab39A interacts with UACA and regulates the retinoic acid-induced neurite morphology of Neuro2A cells. Biochem Biophys Res Commun 435:113–119CrossRefGoogle Scholar
  19. 19.
    Shimizu S, Kondo M, Miyamoto Y, Hayashi M (2002) Foxa (HNF3) up-regulates vitronectin expression during retinoic acid-induced differentiation in mouse neuroblastoma Neuro2a cells. Cell Struct Funct 27:181–188CrossRefGoogle Scholar
  20. 20.
    Lee JY, Tsuchiya R, Miyamoto Y, Hayashi M (1998) A protein reacted with anti-vitronectin antibody accumulates in tumors derived from B16F10 melanoma cells. Cell Struct Funct 23:193–199CrossRefGoogle Scholar
  21. 21.
    Boitard M, Bocchi R, Egervari K, Petrenko V, Viale B, Gremaud S, Zgraggen E, Salmon P, Kiss JZ (2015) Wnt signaling regulates multipolar-to-bipolar transition of migrating neurons in the cerebral cortex. Cell Rep 10:1349–1361CrossRefGoogle Scholar
  22. 22.
    Shah B, Lutter D, Bochenek ML, Kato K, Tsytsyura Y, Glyvuk N, Sakakibara A, Klingauf J, Adams RH, Puschel AW (2016) C3G/Rapgef1 is required in multipolar neurons for the transition to a bipolar morphology during cortical development. PLoS ONE 11:e0154174CrossRefGoogle Scholar
  23. 23.
    Barnat M, Le Friec J, Benstaali C, Humbert S (2017) Huntingtin-mediated multipolar-bipolar transition of newborn cortical neurons is critical for their postnatal neuronal morphology. Neuron 93:99–114CrossRefGoogle Scholar
  24. 24.
    Etienne-Manneville S, Hall A (2003) Cell polarity: Par6, aPKC and cytoskeletal crosstalk. Curr Opin Cell Biol 15:67–72CrossRefGoogle Scholar
  25. 25.
    Betschinger J, Knoblich JA (2004) Dare to be different: asymmetric cell division in Drosophila, C. elegans and vertebrates. Curr Biol 14:R674–685CrossRefGoogle Scholar
  26. 26.
    DeSimone DW, Stepp MA, Patel RS, Hynes RO (1987) The integrin family of cell surface receptors. Biochem Soc Trans 15:789–791CrossRefGoogle Scholar
  27. 27.
    Felding-Habermann B, Cheresh DA (1993) Vitronectin and its receptors. Curr Opin Cell Biol 5:864–868CrossRefGoogle Scholar
  28. 28.
    Neugebauer KM, Emmett CJ, Venstrom KA, Reichardt LF (1991) Vitronectin and thrombospondin promote retinal neurite outgrowth: developmental regulation and role of integrins. Neuron 6:345–358CrossRefGoogle Scholar
  29. 29.
    Isahara K, Yamamoto M (1995) The interaction of vascular endothelial cells and dorsal root ganglion neurites is mediated by vitronectin and heparan sulfate proteoglycans. Brain Res Dev Brain Res 84:164–178CrossRefGoogle Scholar
  30. 30.
    Wang AG, Yen MY, Hsu WM, Fann MJ (2006) Induction of vitronectin and integrin alphav in the retina after optic nerve injury. Mol Vis 12:76–84Google Scholar
  31. 31.
    Katic J, Loers G, Kleene R, Karl N, Schmidt C, Buck F, Zmijewski JW, Jakovcevski I, Preissner KT, Schachner M (2014) Interaction of the cell adhesion molecule CHL1 with vitronectin, integrins, and the plasminogen activator inhibitor-2 promotes CHL1-induced neurite outgrowth and neuronal migration. J Neurosci 34:14606–14623CrossRefGoogle Scholar
  32. 32.
    Yau SY, Li A, Hoo RL, Ching YP, Christie BR, Lee TM, Xu A, So KF (2014) Physical exercise-induced hippocampal neurogenesis and antidepressant effects are mediated by the adipocyte hormone adiponectin. Proc Natl Acad Sci USA 111:15810–15815CrossRefGoogle Scholar
  33. 33.
    Swayne LA, Sorbara CD, Bennett SA (2010) Pannexin 2 is expressed by postnatal hippocampal neural progenitors and modulates neuronal commitment. J Biol Chem 285:24977–24986CrossRefGoogle Scholar
  34. 34.
    Yokosaki Y, Tanaka K, Higashikawa F, Yamashita K, Eboshida A (2005) Distinct structural requirements for binding of the integrins alphavbeta6, alphavbeta3, alphavbeta5, alpha5beta1 and alpha9beta1 to osteopontin. Matrix Biol 24:418–427CrossRefGoogle Scholar
  35. 35.
    Morisaki Y, Niikura M, Watanabe M, Onishi K, Tanabe S, Moriwaki Y, Okuda T, Ohara S, Murayama S, Takao M, Uchida S, Yamanaka K, Misawa H (2016) Selective expression of osteopontin in ALS-resistant motor neurons is a critical determinant of late phase neurodegeneration mediated by matrix metalloproteinase-9. Sci Rep 6:27354CrossRefGoogle Scholar
  36. 36.
    Godin JD, Thomas N, Laguesse S, Malinouskaya L, Close P, Malaise O, Purnelle A, Raineteau O, Campbell K, Fero M, Moonen G, Malgrange B, Chariot A, Metin C, Besson A, Nguyen L (2012) p27(Kip1) is a microtubule-associated protein that promotes microtubule polymerization during neuron migration. Dev Cell 23:729–744CrossRefGoogle Scholar
  37. 37.
    Hnit SS, Xie C, Yao M, Holst J, Bensoussan A, De Souza P, Li Z, Dong Q (2015) p27(Kip1) signaling: transcriptional and post-translational regulation. Int J Biochem Cell Biol 68:9–14CrossRefGoogle Scholar
  38. 38.
    Clement O, Hemming IA, Gladwyn-Ng IE, Qu Z, Li SS, Piper M, Heng JI (2017) Rp58 and p27(kip1) coordinate cell cycle exit and neuronal migration within the embryonic mouse cerebral cortex. Neural Dev 12:8CrossRefGoogle Scholar
  39. 39.
    Rashid T, Banerjee M, Nikolic M (2001) Phosphorylation of Pak1 by the p35/Cdk5 kinase affects neuronal morphology. J Biol Chem 276:49043–49052CrossRefGoogle Scholar
  40. 40.
    Cicero S, Herrup K (2005) Cyclin-dependent kinase 5 is essential for neuronal cell cycle arrest and differentiation. J Neurosci 25:9658–9668CrossRefGoogle Scholar
  41. 41.
    Muley PD, McNeill EM, Marzinke MA, Knobel KM, Barr MM, Clagett-Dame M (2008) The atRA-responsive gene neuron navigator 2 functions in neurite outgrowth and axonal elongation. Dev Neurobiol 68:1441–1453CrossRefGoogle Scholar
  42. 42.
    Le Dreau G, Nicot A, Benard M, Thibout H, Vaudry D, Martinerie C, Laurent M (2010) NOV/CCN3 promotes maturation of cerebellar granule neuron precursors. Mol Cell Neurosci 43:60–71CrossRefGoogle Scholar
  43. 43.
    Milner R, Frost E, Nishimura S, Delcommenne M, Streuli C, Pytela R, Ffrench-Constant C (1997) Expression of alpha vbeta3 and alpha vbeta8 integrins during oligodendrocyte precursor differentiation in the presence and absence of axons. Glia 21:350–360CrossRefGoogle Scholar
  44. 44.
    Blaschuk KL, Frost EE, ffrench-Constant C (2000) The regulation of proliferation and differentiation in oligodendrocyte progenitor cells by alphaV integrins. Development 127:1961–1969Google Scholar
  45. 45.
    Sapir T, Sapoznik S, Levy T, Finkelshtein D, Shmueli A, Timm T, Mandelkow EM, Reiner O (2008) Accurate balance of the polarity kinase MARK2/Par-1 is required for proper cortical neuronal migration. J Neurosci 28:5710–5720CrossRefGoogle Scholar
  46. 46.
    Lin D, Edwards AS, Fawcett JP, Mbamalu G, Scott JD, Pawson T (2000) A mammalian PAR-3-PAR-6 complex implicated in Cdc42/Rac1 and aPKC signalling and cell polarity. Nat Cell Biol 2:540–547CrossRefGoogle Scholar
  47. 47.
    Ohno S (2001) Intercellular junctions and cellular polarity: the PAR-aPKC complex, a conserved core cassette playing fundamental roles in cell polarity. Curr Opin Cell Biol 13:641–648CrossRefGoogle Scholar
  48. 48.
    Etienne-Manneville S, Hall A (2002) Rho GTPases in cell biology. Nature 420:629–635CrossRefGoogle Scholar
  49. 49.
    Goldstein B, Macara IG (2007) The PAR proteins: fundamental players in animal cell polarization. Dev Cell 13:609–622CrossRefGoogle Scholar
  50. 50.
    Xiao Y, Peng Y, Wan J, Tang G, Chen Y, Tang J, Ye WC, Ip NY, Shi L (2013) The atypical guanine nucleotide exchange factor Dock4 regulates neurite differentiation through modulation of Rac1 GTPase and actin dynamics. J Biol Chem 288:20034–20045CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Graduate School of Humanities and SciencesOchanomizu UniversityTokyoJapan
  2. 2.Institute for Human Life InnovationOchanomizu UniversityTokyoJapan

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