Skip to main content

Advertisement

Log in

Regulated Shuttling of Slit-Robo-GTPase Activating Proteins Between Nucleus and Cytoplasm During Brain Development

  • Original Paper
  • Published:
Cellular and Molecular Neurobiology Aims and scope Submit manuscript

Abstract

(1) A Little information exists on the distribution of Slit-Robo-GTPase activating proteins (srGAPs), particularly about their intracellular locations, which may provide further clues to their functions. The purpose of this study is to elucidate the expression patterns of the three srGAPs in wild-type rat brains at adult and various developmental stages, and in the cultured cortical neurons. (2) Immunohistochemical method was applied to detect the distribution and localization of the srGAPs in the normal rat brains at adult and various developmental stages, and in the cultured cortical neurons using the rabbit polyclonal antibodies. (3) Immunohistochemical analysis demonstrated that the three srGAPs were mainly expressed in neurons throughout the brain. More importantly, srGAPs translocated during development by a highly regulated shuttling process between the nucleus and the cytoplasm of neurons and their expression patterns were not overlapping. In cultured cortical neurons srGAPs were found in equal amounts in the cytoplasm, nucleus, in neurites, and growth cones. When neurons were maintained in vitro for longer time, the amount of srGAPs in the nucleus strongly increased. (4) These results suggest that srGAPs are not only involved in the regulation of the Slit-Robo signal transduction, but also in neuronal development and that the translocation of srGAPs is important for their functions.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9

Similar content being viewed by others

References

  • Aznar S, Lacal JC (2001) Rho signals to cell growth and apoptosis. Cancer Lett 165:1–10

    Article  PubMed  CAS  Google Scholar 

  • Brose K, Bland KS, Wang KH, Arnott D, Henzel W, Goodman CS, Tessier-Lavigne M, Kidd T (1999) Slit proteins bind Robo receptors and have an evolutionarily conserved role in repulsive axon guidance. Cell 96:795–806

    Article  PubMed  CAS  Google Scholar 

  • Endris V, Wogatzky B, Leimer U, Bartsch D, Zatyka M, Latif F, Maher ER, Tariverdian G, Kirsch S, Karch D, Rappold GA (2002) The novel Rho-GTPase activating gene MEGAP/srGAP3 has a putative role in severe mental retardation. Proc Natl Acad Sci 99:11754–11759

    Article  PubMed  CAS  Google Scholar 

  • Guan KL, Dixon JE (1991) Eukaryotic proteins expressed in Escherichia coli: an improved thrombin cleavage and purification procedure of fusion proteins with glutathione S-transferase. Anal Biochem 192:262–267

    Article  PubMed  CAS  Google Scholar 

  • Kidd T, Bland KS, Goodman CS (1999) Slit is the midline repellent for the robo receptor in Drosophila. Cell 96:785–794

    Article  PubMed  CAS  Google Scholar 

  • Lanning CC, Daddona JL, Ruiz-Velasco R, Shafer SH, Williams CL (2004) The Rac1 C-terminal polybasic region regulates the nuclear localization and protein degradation of Rac1. J Biol Chem 279:44197–44210

    Article  PubMed  CAS  Google Scholar 

  • Li HS, Chen JH, Wu W, Fagaly T, Zhou L, Yuan W, Dupuis S, Jiang ZH, Nash W, Gick C, Ornitz DM, Wu JY, Rao Y (1999) Vertebrate Slit, a secreted ligand for the transmembrane protein Roundabout, is a repellent for olfactory bulb axons. Cell 96:807–818

    Article  PubMed  CAS  Google Scholar 

  • Madura T, Yamashita T, Kubo T, Tsuji L, Hosokawa K, Tohyama M (2004) Changes in mRNA of Slit-Robo GTPase-activating protein 2 following facial nerve transection. Brain Res Mol Brain Res 123:76–80

    Article  PubMed  CAS  Google Scholar 

  • Marillat V, Cases O, Nguyen-Ba-Charvet KT, Tessier-Lavigne M, Sotelo C, CheDotal A (2002) Spatiotemporal expression patterns of slit and robo genes in the rat brain. J Comp Neurol 442:130–155

    Article  PubMed  Google Scholar 

  • Nguyen Ba-Charvet KT, Brose K, Marillat V, Kidd T, Goodman CS, Tessier-Lavigne M, Sotelo C, Chedotal A (1999) Slit2-mediated chemorepulsion and collapse of developing forebrain axons. Neuron 22:463–473

    Article  PubMed  CAS  Google Scholar 

  • Niclou SP, Jia L, Raper JA (2000) Slit2 is a repellent for retinal ganglion cell axons. J Neurosci 20:4962–4974

    PubMed  CAS  Google Scholar 

  • Perona R, Montaner S, Saniger L, Sanchez-Perez I, Bravo R, Lacal JC (1997) Activation of the nuclear factor-kappaB by Rho, CDC42, and Rac-1 proteins. Genes Dev 11:463–475

    Article  PubMed  CAS  Google Scholar 

  • Sestan N, Artavanis-Tsakonas S, Rakic P (1999) Contact-dependent inhibition of cortical neurite growth mediated by notch signaling. Science 286:741–746

    Article  PubMed  CAS  Google Scholar 

  • Singh RR, Song C, Yang Z, Kumar R (2005). Nuclear localization and chromatin targets of p21-activated kinase 1. J Biol Chem 280:18130–18137

    Article  PubMed  CAS  Google Scholar 

  • Skene JH, Jacobson RD, Snipes GJ, McGuire CB, Norden JJ, Freeman JA (1986) A protein induced during nerve growth (GAP-43) is a major component of growth-cone membranes. Science 233:783–786

    Article  PubMed  CAS  Google Scholar 

  • Soderling SH, Binns KL, Wayman GA, Davee SM, Ong SH, Pawson T, Scott JD (2002) The WRP component of the WAVE-1 complex attenuates Rac-mediated signalling. Nat Cell Biol 4:970–975

    Article  PubMed  CAS  Google Scholar 

  • Stein E, Tessier-Lavigne M (2001) Hierarchical organization of guidance receptors: silencing of netrin attraction by Slit through a Robo/DCC receptor complex. Science 291:1928–1938

    Article  PubMed  CAS  Google Scholar 

  • Suetsugu S, Takenawa T (2003) Translocation of N-WASP by nuclear localization and export signals into the nucleus modulates expression of HSP90. J Biol Chem 278:42515–42523

    Article  PubMed  CAS  Google Scholar 

  • Tanaka T, Nishimura D, Wu RC, Amano M, Iso T, Kedes L, Nishida H, Kaibuchi K, Hamamori Y (2006) Nuclear Rho kinase, ROCK2, targets p300 acetyltransferase. J Biol Chem 281:15320–15329

    Article  PubMed  CAS  Google Scholar 

  • Wang KH, Brose K, Arnott D, Kidd T, Goodman CS, Henzel W, Tessier-Lavigne M (1999) Biochemical purification of a mammalian slit protein as a positive regulator of sensory axon elongation and branching. Cell 96:771–784

    Article  PubMed  CAS  Google Scholar 

  • Whitford KL, Marillat V, Stein E, Goodman CS, Tesssier-Lavigne M, Chedotal A, Ghosh A (2002) Regulation of cortical dendrite development by Slit-Robo interactions. Neuron 33:47–61

    Article  PubMed  CAS  Google Scholar 

  • Williams CL (2003) The polybasic region of Ras and Rho family small GTPases: a regulator of protein interactions and membrane association and a site of nuclear localization signal sequences. Cell Signal 15:1071–1080

    Article  PubMed  CAS  Google Scholar 

  • Wong K, Ren XR, Huang YZ, Xie Y, Liu G, Saito H, Tang H, Wen L, Brady-Kalnay SM, Mei L, Wu JY, Xiong WC, Rao Y (2001) Signal transduction in neuronal migration: roles of GTPase activating proteins and the small GTPase Cdc42 in the Slit-Robo pathway. Cell 107:209–221

    Article  PubMed  CAS  Google Scholar 

  • Wu W, Wong K, Chen J, Jiang Z, Dupuis S, Wu JY, Rao Y (1999) Directional guidance of neuronal migration in the olfactory system by the protein Slit. Nature 400:331–336

    Article  PubMed  CAS  Google Scholar 

  • Yu TW, Hao JC, Lim W, Tessier-Lavigne M, Bargmann CI (2002) Shared receptors in axon guidance: SAX-3/Robo signals via UNC-34/Enabled and a Netrin-independent UNC-40/DCC function. Nat Neurosci 5:1147–1154

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We are grateful for the generous donation of the human KIAA1304, KIAA 0456, and KIAA 0411 clones from the Kazusa DNA Research Institute (Chiba, Japan). The work was supported by the Chinese National Key Project for Basic Research (No. 2003CB515301).

Author information

Authors and Affiliations

Authors

Corresponding authors

Correspondence to Wei-Lin Jin or Gong Ju.

Additional information

Qin Yao and Wei-Lin Jin are contributed equally to this work

Rights and permissions

Reprints and permissions

About this article

Cite this article

Yao, Q., Jin, WL., Wang, Y. et al. Regulated Shuttling of Slit-Robo-GTPase Activating Proteins Between Nucleus and Cytoplasm During Brain Development. Cell Mol Neurobiol 28, 205–221 (2008). https://doi.org/10.1007/s10571-007-9187-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10571-007-9187-4

Keywords

Navigation