Development Genes and Evolution

, Volume 213, Issue 8, pp 382–389 | Cite as

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

  • Jinsong Zhang
  • Shan Bai
  • Carmen Tanase
  • Hideaki Nagase
  • Michael P. SarrasJr.
Original Article


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.


Matrix metalloproteinases MMPs TIMP-2 MMP-2 Zebrafish ECM 


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Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • Jinsong Zhang
    • 1
    • 3
  • Shan Bai
    • 1
  • Carmen Tanase
    • 1
  • Hideaki Nagase
    • 1
    • 2
  • Michael P. SarrasJr.
    • 1
  1. 1.Department of Anatomy and Cell BiologyUniversity of Kansas Medical CenterKansas CityUSA
  2. 2.Kennedy Institute of Rheumatology, Faculty of MedicineImperial College of Science Technology and MedicineLondonUK
  3. 3.Department of Pharmaceutical ChemistryUniversity of KansasLawrenceUSA

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