Event Title

COMPARISON OF EMBRYONIC MALFORMATIONS AFTER FERTILIZATION WITH THALIDOMIDE-TREATED SPERM OR THALIDOMIDE TREATMENT IN CULTURE

Faculty Advisor

Dr. Jan Reichard-Brown

Start Date

23-4-2019 4:20 PM

End Date

23-4-2019 4:40 PM

Description

Thalidomide's mechanism of action as an infamous teratogen remains an unsolved mystery. The literature contains more than 30 theories explaining the human “thalidomide tragedy”. Studies continue to examine the mechanism(s) of action of this drug on both developing embryos and mammalian cells in culture. However, sperm-only exposure to thalidomide and the potential effects on embryonic development remain under research. Sea urchin embryos serve as model systems to study early development with thalidomide. We compared Lytechinus pictus embryos exposed to thalidomide in culture with embryos whose only thalidomide exposure resulted from fertilization with treated sperm. Additionally, we exposed embryos fertilized with thalidomide-treated sperm to thalidomide in the culture dishes as well. Spawning and fertilization followed standard sea urchin protocols. Aliquots of sperm; untreated, treated with seawater, DMSO or thalidomide (0.25nM) were incubated at 17°C for one hour. Thalidomide exposure in culture was 400µM. Examination of 100 embryos per treatment occurred at 24, 48, and 72-hour intervals. Caspase staining for apoptosis used CellEvent® Caspase-3/7 Green ReadyProbes® Reagent. Preliminary data at 48h indicated that fertilization with thalidomide-treated sperm resulted in the highest level of abnormalities, compared to either controls or embryos exposed only in culture. However, embryos fertilized with thalidomide-treated sperm and cultured with thalidomide exhibited more than additive numbers of abnormal embryos. Common malformations at 48 hours included embryonic development out of synchronization with controls, an overabundance of cells, and general aberrant morphology. Preliminary results suggest thalidomide exposure through sperm causes specific areas of cellular death in embryos, opposed to random apoptosis. Our findings indicate that the mode of embryo exposure to thalidomide affects the resulting observed malformations. These experiments suggest that the teratogenic potential of paternal exposure to thalidomide, with or without maternal exposure in humans, needs to be considered.

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Apr 23rd, 4:20 PM Apr 23rd, 4:40 PM

COMPARISON OF EMBRYONIC MALFORMATIONS AFTER FERTILIZATION WITH THALIDOMIDE-TREATED SPERM OR THALIDOMIDE TREATMENT IN CULTURE

Thalidomide's mechanism of action as an infamous teratogen remains an unsolved mystery. The literature contains more than 30 theories explaining the human “thalidomide tragedy”. Studies continue to examine the mechanism(s) of action of this drug on both developing embryos and mammalian cells in culture. However, sperm-only exposure to thalidomide and the potential effects on embryonic development remain under research. Sea urchin embryos serve as model systems to study early development with thalidomide. We compared Lytechinus pictus embryos exposed to thalidomide in culture with embryos whose only thalidomide exposure resulted from fertilization with treated sperm. Additionally, we exposed embryos fertilized with thalidomide-treated sperm to thalidomide in the culture dishes as well. Spawning and fertilization followed standard sea urchin protocols. Aliquots of sperm; untreated, treated with seawater, DMSO or thalidomide (0.25nM) were incubated at 17°C for one hour. Thalidomide exposure in culture was 400µM. Examination of 100 embryos per treatment occurred at 24, 48, and 72-hour intervals. Caspase staining for apoptosis used CellEvent® Caspase-3/7 Green ReadyProbes® Reagent. Preliminary data at 48h indicated that fertilization with thalidomide-treated sperm resulted in the highest level of abnormalities, compared to either controls or embryos exposed only in culture. However, embryos fertilized with thalidomide-treated sperm and cultured with thalidomide exhibited more than additive numbers of abnormal embryos. Common malformations at 48 hours included embryonic development out of synchronization with controls, an overabundance of cells, and general aberrant morphology. Preliminary results suggest thalidomide exposure through sperm causes specific areas of cellular death in embryos, opposed to random apoptosis. Our findings indicate that the mode of embryo exposure to thalidomide affects the resulting observed malformations. These experiments suggest that the teratogenic potential of paternal exposure to thalidomide, with or without maternal exposure in humans, needs to be considered.