Published online 2 September 2005
Article |
Coordinate control of cell cycle regulatory genes in zebrafish development tested by cyclin D1 knockdown with morpholino phosphorodiamidates and hydroxyprolyl-phosphono peptide nucleic acids
1Department of Biochemistry and Molecular Biology, Thomas Jefferson University Philadelphia, PA, USA 2Department of Radiation Oncology, Thomas Jefferson University Philadelphia, PA, USA 3Department of Dermatology and Cutaneous Biology, Thomas Jefferson University Philadelphia, PA, USA 4Department of Microbiology and Immunology, Thomas Jefferson University Philadelphia, PA, USA 5Kimmel Cancer Center, Thomas Jefferson University Philadelphia, PA, USA 6The Wistar Institute Philadelphia, PA, USA 7Jake Gittlen Cancer Research Institute, Pennsylvania State University College of Medicine Hershey, PA, USA
*To whom correspondence should be addressed. Tel: +1 215 955 4578; Fax: +1 215 955 4580; Email: eric{at}tesla.jci.tju.edu
Received July 15, 2005. Revised August 12, 2005. Accepted August 12, 2005.
During early zebrafish (Danio rerio) development zygotic transcription does not begin until the mid-blastula transition (MBT) 
3 h after fertilization. MBT demarcates transition from synchronous short cell cycles of S and M phases exclusively to full cycles encompassing G1 and G2 phases. Transcriptional profiling and RT–PCR analyses during these phases enabled us to determine that this shift corresponds to decreased transcript levels of S/M phase cell cycle control genes (e.g. ccna2, ccnb1, ccnb2 and ccne) and increased transcript levels of ccnd1, encoding cyclin D1, and orthologs of p21 (p21-like) and retinoblastoma (Rb-like 1). To investigate the regulation of this process further, the translation of ccnd1 mRNA, a G1/S checkpoint control element, was impaired by microinjection of ccnd1-specific morpholino phosphorodiamidate (MO) 20mer or hydroxyprolyl-phosphono peptide nucleic acid (HypNA-pPNA) 16mer antisense oligonucleotides. The resulting downregulation of cyclin D1 protein resulted in microophthalmia and microcephaly, but not lethality. The phenotypes were not seen with 3-mismatch MO 20mers or 1-mismatch HypNA-pPNA 16mers, and were rescued by an exogenous ccnd1 mRNA construct with five mismatches. Collectively, these results indicate that transcription of key molecular determinants of asynchronous cell cycle control in zebrafish embryos commences at MBT and that the reduction of cyclin D1 expression compromises zebrafish eye and head development.
Present addresses: Kevin T. Duffy, AstraZeneca, Wayne, PA 19087, USA
Laszlo Kari, Rocky Mountain Laboratories, NIAID, Hamilton, MT 59840, USA
Chang-Gong Liu, Institute of Genetics, Ohio State University, Columbus, OH 43210, USA
Steven A. Farber, Department of Embryology, Carnegie Institution of Washington, Baltimore, MD 21218, USA
The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  J. Bessa, M. J. Tavares, J. Santos, H. Kikuta, M. Laplante, T. S. Becker, J. L. Gomez-Skarmeta, and F. Casares meis1 regulates cyclin D1 and c-myc expression, and controls the proliferation of the multipotent cells in the early developing zebrafish eye Development, March 1, 2008; 135(5): 799 - 803. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Feitsma, M. C. Leal, P. B. Moens, E. Cuppen, and R. W. Schulz Mlh1 Deficiency in Zebrafish Results in Male Sterility and Aneuploid as Well as Triploid Progeny in Females Genetics, April 1, 2007; 175(4): 1561 - 1569. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. A. Geiger, S. E. Parker, A. P. Beothy, J. A. Tucker, M. C. Mullins, and G. D. Kao Zebrafish as a "Biosensor"? Effects of Ionizing Radiation and Amifostine on Embryonic Viability and Development Cancer Res., August 15, 2006; 66(16): 8172 - 8181. [Abstract] [Full Text] [PDF]
|
|