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Updated: Jul 30, 2026
In vivo-like Organotypic Murine Retinal Wholemount Culture
Published on: January 11, 2010
A new model for assessing proteolysis in the intact mouse lens in organ culture
A Petersen1, M Zetterberg, J Sjöstrand
1Institute of Anatomy and Cell Biology, University of Göteborg, Göteborg, Sweden. Anne.Petersen@anatcell.gu.se
Purpose:
To develop a new method to investigate proteolysis in the intact lens in organ culture.
Methods:
Intact mouse lenses were assayed at regular intervals for proteolytic activity using fluorogenic peptide substrates +/- addition of ionomycin. Specific inhibitors were used to determine the activity of calpains, the proteasome and acid lysosomal enzymes.
Results:
Significant levels of proteolytic activity were present in the intact lens. Proteolysis was stimulated by ionomycin. Preincubation with an inhibitor to the proteasome significantly decreased proteolysis whereas inhibitors of calpain and acid lysosomal enzymes did not.
Conclusion:
This study indicates that in the intact mouse lens in culture, the proteasome is an important protease. Its activity is at least partially regulated by calcium.
