Abstract
Microtubules play important roles in mitotic spindle assembly and chromosome segregation to maintain normal cell cycle progression. A number of microtubule-associated proteins have been identified in epithelial and neural cell cultures; however, their physiological significance is not well characterized due to the lack of appropriate in vivo animal models. Nucleolar spindle-associated protein (NuSAP) is a microtubule-binding protein and is reported to be involved in mitosis by cell culture studies. In this report, we identified the zebrafish homologue of human NuSAP and investigated its expression profile and functions. Using in situ hybridization, we demonstrated that transcripts of zebrafish nusap1 are specifically expressed in the retina, forebrain, hindbrain and neural crest. When the in vivo expression of nusap1 was knocked down through antisense oligonucleotide morpholino technology, the morphants of nusap1 showed impaired morphogenesis in the trunk and yolk extension, implying the involvement of Nusap1 in cell migration. Mechanistic studies revealed that nusap1 morphants have an altered expression pattern of neural crest markers crestin and sox9b, but normal expression of blood vessel and notochord markers gata1 and shh. In addition, nusap1 mRNA injection caused serious apoptosis in retina and hindbrain tissue, and these phenotypes can be rescued by co-injection of morpholino against nusap1. These observations not only suggest a role for Nusap1 in connecting apoptosis with cell migration, but also provide strong evidences that Nusap1 is potentially involved in morphogenesis in vertebrates.
Article PDF
Similar content being viewed by others
References
Chen, J., Ng, S.M., Chang, C., Zhang, Z., Bourdon, J.C., Lane, D.P., and Peng, J. (2009). p53 isoform delta113p53 is a p53 target gene that antagonizes p53 apoptotic activity via BclxL activation in zebrafish. Genes Dev 23, 278–290.
Herzog, W., Zeng, X., Lele, Z., Sonntag, C., Ting, J.-W., Chang, C.-Y., and Hammerschmidt, M. (2003). Adenohypophysis formation in the zebrafish and its dependence on sonic hedgehog. Dev Biol 254, 36–49.
Hetzer, M., Bilbao-Cort, D., Walther, T.C., Gruss, O.J., and Mattaj, I.W. (2000). GTP hydrolysis by Ran is required for nuclear envelope assembly. Mol Cell 5, 1013–1024.
Huang, H., Lu, F.-I., Jia, S., Meng, S., Cao, Y., Wang, Y., Ma, W., Yin, K., Wen, Z., Peng, J., et al. (2007). Amotl2 is essential for cell movements in zebrafish embryo and regulates c-Src translocation. Development 134, 979–988.
Kokkinakis, D.M., Liu, X., and Neuner, R.D. (2005). Modulation of cell cycle and gene expression in pancreatic tumor cell lines by methionine deprivation (methionine stress): implications to the therapy of pancreatic adenocarcinoma. Mol Cancer Ther 4, 1338–1348.
Li, L., Zhou, Y., Sun, L., Xing, G., Tian, C., Sun, J., Zhang, L., and He, F. (2007). NuSAP is degraded by APC/C-Cdh1 and its overexpression results in mitotic arrest dependent of its microtubules’ affinity. Cell Signal 19, 2046–2055.
Li, Q., Lin, S., Wang, X., Lian, G., Lu, Z., Guo, H., Ruan, K., Wang, Y., Ye, Z., Han, J., et al. (2009). Axin determines cell fate by controlling the p53 activation threshold after DNA damage. Nat Cell Biol 11, 1128–1134.
Maves, L., Waskiewicz, A.J., Paul, B., Cao, Y., Tyler, A., Moens, C.B., and Tapscott, S.J. (2007). Pbx homeodomain proteins direct Myod activity to promote fast-muscle differentiation. Development 134, 3371–3382.
Meng, A., Jessen, J.R., and Lin, S. (1999). Transgenesis. Methods Cell Biol 60, 133–148.
Merdes, A., Ramyar, K., Vechio, J.D., and Cleveland, D.W. (1996). A complex of NuMA and cytoplasmic dynein is essential for mitotic spindle assembly. Cell 87, 447–458.
Ornitz, D.M. (2005). FGF signaling in the developing endochondral skeleton. Cytokine Growth Factor Rev 16, 205–213.
Raemaekers, T., Ribbeck, K., Beaudouin, J., Annaert, W., Van Camp, M., Stockmans, I., Smets, N., Bouillon, R., Ellenberg, J., and Carmeliet, G. (2003). NuSAP, a novel microtubule-associated protein involved in mitotic spindle organization. J Cell Biol 162, 1017–1029.
Ribbeck, K., Groen, A.C., Santarella, R., Bohnsack, M.T., Raemaekers, T., Kocher, T., Gentzel, M., Gorlich, D., Wilm, M., Carmeliet, G., et al. (2006). NuSAP, a mitotic RanGTP target that stabilizes and cross-links microtubules. Mol Biol Cell 17, 2646–2660.
Ribbeck, K., Raemaekers, T., Carmeliet, G., and Mattaj, I.W. (2007). A role for NuSAP in linking microtubules to mitotic chromosomes. Curr Biol 17, 230–236.
Robu, M.E., Larson, J.D., Nasevicius, A., Beiraghi, S., Brenner, C., Farber, S.A., and Ekker, S.C. (2007). p53 activation by knockdown technologies. PLoS Genet 3, e78.
Vousden, K.H., and Lu, X. (2002). Live or let die: the cell’s response to p53. Nat Rev Cancer 2, 594–604.
Warga, R.M., Kane, D.A., and Ho, R.K. (2009). Fate mapping embryonic blood in zebrafish: multi- and unipotential lineages are segregated at gastrulation. Dev Cell 16, 744–755.
Xiong, B., Rui, Y., Zhang, M., Shi, K., Jia, S., Tian, T., Yin, K., Huang, H., Lin, S., Zhao, X., et al. (2006). Tob1 controls dorsal development of zebrafish embryos by antagonizing maternal beta-catenin transcriptional activity. Dev Cell 11, 225–238.
Yudin, D., and Fainzilber, M. (2009). Ran on tracks-cytoplasmic roles for a nuclear regulator. J Cell Sci 122, 587–593.
Author information
Authors and Affiliations
Corresponding authors
Electronic supplementary material
Rights and permissions
About this article
Cite this article
Nie, J., Wang, H., He, F. et al. Nusap1 is essential for neural crest cell migration in zebrafish. Protein Cell 1, 259–266 (2010). https://doi.org/10.1007/s13238-010-0036-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13238-010-0036-8