Abstract
Gelatinase A, also called matrix metalloproteinase 2 (MMP-2), belongs to the matrix metalloproteinase (MMP) family. MMP-2 cleaves type IV collagen, denatured collagen (gelatin), and other extracellular matrix (ECM) components. MMP-2 has been reported to be involved in a number of biological and pathological processes, but previous studies have not indicated that its expression is essential for early embryogenesis. In the current study, we have utilized zebrafish as a developmental model to study the role of MMP-2 during embryogenesis. We have successfully isolated a zebrafish MMP-2 (zMMP-2) homologue showing over 80% identity and over 90% similarity to its human counterpart. In situ analysis showed that zMMP-2 was expressed as early as the one-cell stage implying a maternal origin during oogenesis, and embryos continued to express zMMP-2 through at least the 72-h stage of development. RT-PCR analysis confirmed the in situ expression pattern and gelatin zymography indicated that a metalloproteinase with the same gel mobility as vertebrate MMP-2 was present in zebrafish embryos. Injection of zMMP-2 antisense morpholino oligonucleotides into 1- to 4-cell embryos resulted in a truncated axis, monitored through 72 h of development indicating that this metalloproteinase plays an important role in zebrafish embryogenesis. Monpholino-induced alterations in development began to be observed at 12 h of embryogenesis based on morphological and axis marker studies. The results obtained in zebrafish are in contrast to murine knockout studies that indicate that MMP-2 does not have a major role in mouse embryogenesis.
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Acknowledgements
The authors wish to thank Li Li for technical assistance and 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 used in this study. The work was supported by NIH Grants AR39189, DK61373, American Heart Grant 0051346Z, and The Wellcome Trust Grant 057508.
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Zhang, J., Bai, S., Zhang, X. et al. The expression of gelatinase A (MMP-2) is required for normal development of zebrafish embryos. Dev Genes Evol 213, 456–463 (2003). https://doi.org/10.1007/s00427-003-0346-4
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DOI: https://doi.org/10.1007/s00427-003-0346-4