Inhibition of Plk1 induces mitotic infidelity and embryonic growth defects in developing zebrafish embryos

Kilhun Jeong; Jae-Yeon Jeong; Hae-Ock Lee; Eunhee Choi; Hyunsook Lee

doi:10.1016/j.ydbio.2010.06.004

Inhibition of Plk1 induces mitotic infidelity and embryonic growth defects in developing zebrafish embryos

Dev Biol. 2010 Sep 1;345(1):34-48. doi: 10.1016/j.ydbio.2010.06.004. Epub 2010 Jun 8.

Authors

Kilhun Jeong¹, Jae-Yeon Jeong, Hae-Ock Lee, Eunhee Choi, Hyunsook Lee

Affiliation

¹ Department of Biological Sciences and Institute of Molecular Biology and Genetics, College of Natural Sciences, Seoul National University, 599, Gwanak-Ro, Gwanak-Gu, Seoul 151-742, Korea.

PMID: 20553902
DOI: 10.1016/j.ydbio.2010.06.004

Abstract

Polo-like kinase 1 (Plk1) is central to cell division. Here, we report that Plk1 is critical for mitosis in the embryonic development of zebrafish. Using a combination of several cell biology tools, including single-cell live imaging applied to whole embryos, we show that Plk1 is essential for progression into mitosis during embryonic development. Plk1 morphant cells displayed mitotic infidelity, such as abnormal centrosomes, irregular spindle assembly, hypercondensed chromosomes, and a failure of chromosome arm separation. Consequently, depletion of Plk1 resulted in mitotic arrest and finally death by 6days post-fertilization. In comparison, Plk2 or Plk3 morphant embryos did not display any significant abnormalities. Treatment of embryos with the Plk1 inhibitor, BI 2536, caused a block in mitosis, which was more severe when used to treat plk1 morphants. Finally, using an assay to rescue the Plk1 morphant phenotype, we found that the kinase domain and PBD domains are both necessary for Plk1 function in zebrafish development. Our studies demonstrate that Plk1 is required for embryonic proliferation because its activity is crucial for mitotic integrity. Furthermore, our study suggests that zebrafish will be an efficient and economical in vivo system for the validation of anti-mitotic drugs.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Animals
Animals, Genetically Modified
Apoptosis
Cell Cycle Proteins / antagonists & inhibitors
Cell Cycle Proteins / genetics*
Cell Cycle Proteins / metabolism
Cell Proliferation
Chromosome Segregation / drug effects
Embryo, Nonmammalian / cytology
Embryo, Nonmammalian / embryology
Embryo, Nonmammalian / metabolism*
Gene Expression Regulation, Developmental
Gene Knockdown Techniques
Green Fluorescent Proteins / genetics
Green Fluorescent Proteins / metabolism
Histones / genetics
Histones / metabolism
In Situ Hybridization
Kinetics
Microscopy, Video / methods
Mitosis / drug effects
Molecular Sequence Data
Polo-Like Kinase 1
Protein Serine-Threonine Kinases / antagonists & inhibitors
Protein Serine-Threonine Kinases / genetics*
Protein Serine-Threonine Kinases / metabolism
Proto-Oncogene Proteins / antagonists & inhibitors
Proto-Oncogene Proteins / genetics*
Proto-Oncogene Proteins / metabolism
Pteridines / pharmacology
Reverse Transcriptase Polymerase Chain Reaction
Sequence Homology, Amino Acid
Zebrafish / embryology
Zebrafish / genetics
Zebrafish Proteins / antagonists & inhibitors
Zebrafish Proteins / genetics*
Zebrafish Proteins / metabolism

Substances

BI 2536
Cell Cycle Proteins
Histones
Proto-Oncogene Proteins
Pteridines
Zebrafish Proteins
Green Fluorescent Proteins
Protein Serine-Threonine Kinases