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ARS Home » Research » Publications at this Location » Publication #85364
Title: MICROFILAMENT STABILITY DURING SWINE EMBRYO CRYOPRESERVATION.

Author
item Dobrinsky, John
item Long, Charles
item Johnson, Lawrence

Submitted to: Biology of Reproduction
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 9/30/1999
Publication Date: N/A
Citation: N/A

Interpretive Summary: Documenting cell damage during cryopreservation provides useful information for understanding embryonic sensitivity and cellular response to cryopreservation. The cytoskeleton of embryos is essential for controlling shape, intracellular movement, organelle transport, chromosome segregation and communication within the cell. Many cryoprotective agents function to depolymerize cytoskeletal components prior to cooling and may be toxic to cells. The objective of this study was to document microfilament (MF) alterations using confocal microscopy during swine embryo vitrification and the use of cytochalasin-b, a MF inhibitor, during cryopreservation. Vitrified morulae/early blastocysts displayed MF disruptions and lacked developmental competence after cryopreservation; whereas, hatched blastocysts displayed variable MF disruption and developmental competence. Treatment with cytochalasin-b did not improve morulae/early blastocyst viability after vitrification; however, it significantly improved survival of expanded and hatched blastocysts. This study shows that the pig embryo cytoskeleton can be affected by vitrification, and that microfilament depolymerization prior to vitrification improves blastocyst development. Researchers will use this information and scientific approach to studying cellular disruption during cryopreservation to improve embryo cryopreservation methodology with the eventual goal of producing live offspring after transfer of cryopreserved swine embryos.

Technical Abstract: Documenting cellular damage during cryopreservation provides useful information for understanding embryonic sensitivity and cellular response to cryopreservation. The cytoskeleton of mammalian embryos is essential for controlling shape, intracellular movement, organelle transport and chromosome segregation while providing a pathway of communication within the cell. Many cryoprotective agents function to depolymerize cytoskeletal components prior to cooling and may be toxic to cells. The objective of this study was to document microfilament (MF) alterations during swine embryo vitrification and the use of cytochalasin-b, a MF inhibitor, during cryopreservation. Microfilament polymerization was monitored before, during and after vitrification. Vitrified morulae/early blastocysts displayed MF disruptions and lacked developmental competence after cryopreservation; whereas, hatched blastocysts displayed variable MF disruption and developmental competence. Cytochalasin-b caused a reduced fluorescence intensity of microfilaments as revealed with confocal microscopy. Treatment with cytochalasin-b did not improve morulae/early blastocyst viability after vitrification; however, it significantly improved survival of expanded (2-fold) and hatched blastocysts (3-fold). This study shows that the pig embryo cytoskeleton can be affected by vitrification, and that microfilament depolymerization prior to vitrification improves blastocyst development.