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Characterization of matrix metalloproteinase in flatfish embryo undergoing formation of lenses
1Division of Physiology and Molecular Biology, National Research Institute of Fisheries Science, Yokohama, Japan. aranishi@nrifs.affrc.go.jp
Abstract:
Members of the matrix metalloproteinase (MMP) family are responsible for breakdown of extracellular matrix components involved in morphogenetic remodeling of animal embryogenesis. The highly sensitive assay of MMP using synthetic fluorescence-quenching substrate was employed to detect and to characterize a veiled MMP activity expressed in Japanese flounder embryos undergoing formation of lenses. The MMP activity was enhanced in proportion to increasing protein amounts of the embryonic lysate over 5 microg, and this reaction was proceeded in a time-dependent manner and with increasing substrate concentrations. Almost 2-fold increase in the embryonic MMP activity occurred by treatment with 4-aminophenylmercuric acetate, but the activity was markedly suppressed by metal chelating reagents. These enzymatic characteristics are apparently consistent with those of mammalian embryonic MMPs, particularly MMP-9. The characterized MMP activity was highly expressed at the specificstage during embryogenesis, indicating that this MMP may be involved in formation of lenses.
Insights
Matrix metalloproteinase (MMP) activity was detected in Japanese flounder embryos, suggesting a role in lens formation. This embryonic MMP activity shares characteristics with mammalian MMP-9.
Area of Science:
- Developmental Biology
- Biochemistry
- Molecular Biology
Background:
- Matrix metalloproteinases (MMPs) are crucial for extracellular matrix remodeling during embryonic development.
- Understanding MMP roles in specific developmental processes, like lens formation, is vital.
Purpose of the Study:
- To detect and characterize matrix metalloproteinase (MMP) activity during Japanese flounder embryogenesis.
- To investigate the potential involvement of MMPs in lens formation.
Main Methods:
- Utilized a sensitive assay with a synthetic fluorescence-quenching substrate to measure MMP activity.
- Analyzed MMP activity in embryonic lysates based on protein concentration, time, and substrate concentration.
- Investigated the effects of 4-aminophenylmercuric acetate and metal chelating reagents on MMP activity.
Main Results:
- A veiled MMP activity was detected and characterized in Japanese flounder embryonic lysates.
- MMP activity showed a dose-dependent relationship with protein concentration and was time-dependent.
- Activity increased with substrate concentration, was enhanced by 4-aminophenylmercuric acetate, and inhibited by metal chelators.
- Enzymatic properties were consistent with mammalian embryonic MMPs, particularly MMP-9.
- The characterized MMP activity was highly expressed during a specific embryogenesis stage.
Conclusions:
- The detected MMP activity in Japanese flounder embryos is likely involved in lens formation.
- This embryonic MMP may be functionally analogous to mammalian MMP-9, highlighting conserved roles in development.