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The expression of tissue inhibitor of metalloproteinase 2 (TIMP-2) is required for normal development of zebrafish embryos

Development Genes and Evolution volume 213, pages 382–389 (2003)Cite this article

Abstract

MMP activities are controlled by a combination of proteolytic pro-enzyme activation steps and inhibition by endogenous inhibitors like α2-macroglobulin and the tissue inhibitors of metalloproteinases (TIMPs). TIMPs are the key inhibitors in tissue. The expression of both MMPs and TIMPs is controlled during tissue remodeling to maintain a balance in the turnover of extracellular matrix. Disruption of this balance may result in a broad spectrum of diseases. Additionally, TIMP-2 has been reported to have growth factor activities. To further study the function of TIMP-2 in development, we utilized zebrafish as an experimental model system. We have successfully isolated a TIMP-2 homologue from zebrafish (zTIMP-2). This zebrafish TIMP-2 showed high similarity to human TIMP-2 with all critical features conserved. Whole-mount in situ analysis showed that zTIMP-2 was expressed as early as the one-cell stage indicating a maternal origin. This expression continued through later stages of development. RT-PCR analysis confirmed the early expression pattern from the 16-cell stage through blastula, gastrula and 24-h stages. In addition, at the protein level, immunoreactive zTIMP-2 was detected using antibody against recombinant human TIMP-2. RFP-reporter analysis indicated that TIMP-2 can be secreted into the extracellular space where ECM is forming. Functional studies showed that the balance of TIMP-2 expression is important to normal development as reflected by the fact that both blockage of TIMP-2 translation using antisense morpholino oligonculeotides or increased translation of TIMP-2 using a mRNA microinjection approach resulted in abnormal zebrafish development. This is in contrast to murine knockout studies that indicate that TIMP-2 does not have a major role in mouse embryogenesis.

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Acknowledgements

The authors wish to thank Eileen Roach for assistance with preparation of the figures used in this article. The authors also wish to thank Dr. Michael Rebagliati of the University of Iowa Medical Center for providing the axis specification markers. The work was supported by NIH Grants AR39189, AR40994, DK61373, and American Heart Grant 0051346Z.

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Author notes

  1. Jinsong Zhang

    Present address: Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, KS, USA

Affiliations

  1. Department of Anatomy and Cell Biology, University of Kansas Medical Center, 3901 Rainbow Blvd., Kansas City, KS 66160, USA

    Jinsong Zhang, Shan Bai, Carmen Tanase, Hideaki Nagase & Michael P. Sarras Jr.

  2. Kennedy Institute of Rheumatology, Faculty of Medicine, Imperial College of Science Technology and Medicine, London, UK

    Hideaki Nagase

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  1. Jinsong Zhang
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  2. Shan Bai
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  3. Carmen Tanase
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  4. Hideaki Nagase
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  5. Michael P. Sarras Jr.
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Correspondence to Michael P. Sarras Jr..

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Zhang, J., Bai, S., Tanase, C. et al. The expression of tissue inhibitor of metalloproteinase 2 (TIMP-2) is required for normal development of zebrafish embryos. Dev Genes Evol 213, 382–389 (2003). https://doi.org/10.1007/s00427-003-0333-9

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  • DOI: https://doi.org/10.1007/s00427-003-0333-9

Keywords

  • Matrix metalloproteinases
  • MMPs
  • TIMP-2
  • MMP-2
  • Zebrafish ECM