Volume 4 Supplement 1

Proceedings of the Second International Cilia in Development and Disease Scientific Conference (2014)

Open Access

Role of outer dense fiber of sperm tails 2-like (ODF2L) protein in ciliation in mammalian cells and in zebrafish

  • P De Saram1,
  • C Wilkinson1 and
  • J Murdoch1
Cilia20154(Suppl 1):P32

https://doi.org/10.1186/2046-2530-4-S1-P32

Published: 13 July 2015

Background

The centrosome is a subcellular organelle which plays an important role within the cell as the microtubule organising centre (MTOC) and many other cellular functions. In a quiescent cell, centrioles can migrate to the apical surface of the cell and nucleate hair-like projections called cilia and flagella. The stages of ciliogenesis require a large number of proteins and complexes to be trafficked through to the cilium and are carefully coordinated by the centrosome and centrosome-associated proteins. Here, we studied centrosome proteins named outer dense fiber-2 like (ODF2L) to establish their role in ciliogenesis in vitro and in zebrafish (Danio rerio).

Results

We identified the ODF2L as a satellite protein in proliferating cells by using immunofluorescent-labeling and expression of this protein is changed when the ciliation is induced. When ciliation induced, ODF2L seems to localise into the centrioles or completely diminish the expression in hTERT-RPE-1 cells. When lysate of the quiescent cells was analysed for the ODF2L expression revealed the complete removal of the protein. When ODF2L was over expressed, hTERT-RPE-1, were unable to ciliate even after initiation of ciliation by serum depravation. Furthermore, knocking down of ODF2L with SiRNA in hTERT-RPE-1 cells resulted in four times more cilia cultured in serum supplemented media. In zebrafish depleting the transcript by using anti-sensing morpholinos resulted in the phenotype of varied development and curved back with ectopic otoliths which resembles other ciliary function compromised phenotypes observed when the known ciliary components were depleted. Furthermore, whole mount immunoflorescent studies of morphants revealed shortened cilia in the pronephros.

Conclusion

ODF2L may negatively regulate ciliation by changing the expression and its localisation within mammalian cells however in zebrafish exact function is yet to be discovered.

Authors’ Affiliations

(1)
Biological Sciences, Royal Holloway University of London

Copyright

© Saram et al. 2015

This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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