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Brain Structure and Function

, Volume 219, Issue 1, pp 105–118 | Cite as

Neuritin 1 promotes neuronal migration

  • Arianna Zito
  • Daniele Cartelli
  • Graziella Cappelletti
  • Anna Cariboni
  • William Andrews
  • John Parnavelas
  • Angelo PolettiEmail author
  • Mariarita Galbiati
Original Article

Abstract

Neuritin 1 (Nrn1 or cpg15-1) is an activity-dependent protein involved in synaptic plasticity during brain development, a process that relies upon neuronal migration. By analyzing Nrn1 expression, we found that it is highly expressed in a mouse model of migrating immortalized neurons (GN11 cells), but not in a mouse model of non-migrating neurons (GT1-7 cells). We thus hypothesized that Nrn1 might control neuronal migration. By using complementary assays, as Boyden’s microchemotaxis, scratch-wounding and live cell imaging, we found that GN11 cell migration is enhanced when Nrn1 is overexpressed and decreased when Nrn1 is silenced. The effects of Nrn1 in promoting neuronal migration have been then confirmed ex vivo, on rat cortical interneurons, by Boyden chamber assays and focal electroporation of acute embryonic brain slices. Furthermore, we found that Nrn1 level modulation affects GN11 cell morphology. The process is also paralleled by Nrn1-induced α-tubulin post-translational modifications, a well-recognized marker of microtubule stability. Altogether, the data demonstrate a novel function of Nrn1 in promoting migration of neuronal cells and indicate that Nrn1 levels impact on microtubule stability.

Keywords

Neuritin Cpg15 Cell migration GnRH neurons Ganglionic eminence Microtubule 

Notes

Acknowledgments

Telethon-Italy to AP (GGP07063); Fondazione CARIPLO to AP (2008–2307); AriSLA, Italy to AP; Italian Ministry of Labour, Health and Social Affairs to AP (Convenzione Fondazione Mondino/UNIMI); Fondation Thierry Latran, France to AP; Italian Ministry of University and Research to MG; “Dote ricerca”, FSE, Regione Lombardia, to DC and to AP.

Supplementary material

429_2012_487_MOESM1_ESM.tif (1.1 mb)
Controls for the silencing effect on Nrn1. A) RT-qPCR analysis of Nrn1 mRNA levels in control, silenced (shRNA-Nrn1) and scramble transfected (shRNA-Scr) GN11 cells. *p<0.01 vs. control and shRNA-Scr. B) Immunoblot and densitometric analyses of Nrn1 in silenced GN11 cells. *p<0.01 vs. control and shRNA-Scr. Supplementary material 1 (TIFF 1166 kb)
429_2012_487_MOESM2_ESM.tif (1.1 mb)
Controls for the overexpression of Nrn1. A) RT-qPCR analysis of Nrn1 mRNA levels in control and Nrn1 overexpressing (pIRES-Nrn1) GN11 cells. *p<0.01 vs. Control. B) Immunoblot of Nrn1 overexpressing GN11 cells. Membranes have been processed with anti-Nrn1 and anti-Flag antibodies. Supplementary material 2 (TIFF 1149 kb)

This movie shows shRNA-Scr control GN11 cells undergoing migration. The cells were cultured in serum free medium for 3 hours and, then, recorded for 8 h. Time-lapse microscopy images were taken at 10 minutes intervals. The neurons, both spindle shaped or multipolar, exhibit a spontaneous migratory ability. Supplementary material 3 (MPG 324 kb)

This movie shows Nrn1 silenced GN11 cells undergoing migration. The cells were cultured in serum free medium for 3 hours and, then, recorded for 8 h. Time-lapse microscopy images were taken at 10 minutes intervals. The neurons do not show any appreciable migration at all the monitored time even if displaying a spindle-shaped morphology. Supplementary material 4 (MPG 408 kb)

This movie shows Nrn1 overexpressing GN11 cells undergoing migration. The cells were cultured in serum free medium for 3 hours and, then, recorded for 8 h. Time-lapse microscopy images were taken at 10 minutes intervals. The neurons migrate considerably, moving faster and at distance longer than that observed for control cells. Supplementary material 5 (MPG 490 kb)

This movie shows pEGFP-N1 transfected control GN11 cells undergoing migration. The cells were cultured in serum free medium for 3 hours and, then, recorded for 8 h. Time-lapse microscopy images were taken at 10 minutes intervals. The neurons, both spindle shaped or multipolar, exhibit a spontaneous migratory ability. Supplementary material 6 (MPG 406 kb)

429_2012_487_MOESM7_ESM.doc (30 kb)
Supplementary material 7 (DOC 30 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2012

Authors and Affiliations

  • Arianna Zito
    • 1
  • Daniele Cartelli
    • 2
  • Graziella Cappelletti
    • 2
  • Anna Cariboni
    • 1
    • 3
  • William Andrews
    • 3
  • John Parnavelas
    • 3
  • Angelo Poletti
    • 1
    • 4
    Email author
  • Mariarita Galbiati
    • 1
    • 4
  1. 1.Dipartimento di Scienze Farmacologiche e Biomolecolari and Centre of Excellence on Neurodegenerative DiseasesUniversità degli Studi di Milano, ItaliaMilanItaly
  2. 2.Dipartimento di BioscienzeUniversità degli Studi di Milano, ItaliaMilanItaly
  3. 3.Department of Cell and Developmental BiologyUniversity College LondonLondonUK
  4. 4.Inter-University Research Centre on the Molecular Basis of Neurodegenerative DiseasesRomeItaly

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