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Updated: Jul 7, 2026

Analysis of Apoptosis in Zebrafish Embryos by Whole-mount Immunofluorescence to Detect Activated Caspase 3
Published on: December 20, 2013
Relationship between group II caspase activity of bovine preimplantation embryos and capacity for hatching
Frank Dean Jousan1, Luiz Augusto de Castro E Paula, Amber Marie Brad
1Department of Animal Sciences, University of Florida, Gainesville, FL 32611-0910, USA.
Abstract:
The occurrence of apoptosis in a fraction of blastomeres in the preimplantation embryo is well known but the consequences of this phenomenon for the developmental potential of the blastocyst has not been well established. Here we demonstrate that blastocysts with low amounts of activated group II caspase activity have increased potential for development to the hatched blastocyst stage. Bovine blastocysts produced in vitro were assayed using a non-invasive fluoregenic substrate that is cleaved by activated group II caspases (i.e., caspase-2, -3 and -7). Subsequently, blastocysts were cultured until Day 10 post-insemination and the proportion undergoing hatching determined. In Experiment 1, blastocysts were cultured without respect to stage of development (expanded or non-expanded); blastocysts classified as having low caspase activity had higher hatching rates than blastocysts with medium or high caspase activity. In Experiment 2, embryos were categorized as nonexpanded or expanded blastocysts. Caspase activity was lower and hatching rate higher for expanded blastocysts than for nonexpanded blastocysts. For nonexpanded blastocysts, embryos classified as having low caspase activity had higher hatching rates as compared to embryos with medium or high caspase activity. In conclusion, the capacity for blastocysts to undergo further development is related to degree of group II caspase activity. Conditions that enhance the incidence of apoptosis in blastocysts may reduce developmental competence. In addition, determination of caspase activity may be useful for selection of embryos for transfer into recipients.

