Cryopreservation of Musca domestica (Diptera: Muscidae) embryos.
Prior studies on cryopreserving embryos of several non-drosophilid flies established that two Drosophila melanogaster embryo cryopreservation protocols were not directly suitable for use with these species. This paper describes our work on developing a protocol for cryopreservation of embryos of the housefly, Musca domestica. Significant progress was made when permeabilization of the vitelline membrane was optimized, a vitrification solution containing ethylene glycol, polyethylene glycol, and trehalose was formulated, and when cooling and recovery of the cryopreservation protocol included a step which passed the embryos through liquid nitrogen vapor. More than 70% of housefly embryos withstand treatments of dechorionation, permeabilization, loading with cryoprotectant, and dehydration in vitrification solution, but the cooling, warming, and poststorage rearing steps still cause a considerable reduction in survival. About 53% of the vitrified M. domestica embryos hatched into larvae. Relative to the percentage of the control adult emergence, about 13% of the embryos stored in liquid nitrogen developed into fertile adults. Hatching of the F(1) progeny of adults having been cryopreserved as embryos was similar to control levels.
Wang
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WB
Department of Plant Pathology, North Dakota State University, Fargo, North Dakota 58105, USA. wangwb@fargo.ars.usda.gov
Leopold
R A
RA
Nelson
D R
DR
Freeman
T P
TP
D019840:2-Propanol / Q000494:pharmacology; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D015925:Cryopreservation; D003451:Cryoprotective Agents / Q000494:pharmacology; D003470:Culture Media / Q000494:pharmacology; D003890:Desiccation; D004625:Embryo, Nonmammalian / Q000737:chemistry / Q000187:drug effects; D019855:Ethylene Glycol / Q000494:pharmacology; D005260:Female; D005298:Fertility; D006536:Heptanes / Q000494:pharmacology; D006586:Hexanes / Q000494:pharmacology; D006793:Houseflies / Q000196:embryology*; D008055:Lipids / Q000032:analysis; D008297:Male; D009584:Nitrogen; D010539:Permeability / Q000187:drug effects; D011092:Polyethylene Glycols / Q000494:pharmacology; D012996:Solutions; D013696:Temperature; D014199:Trehalose / Q000494:pharmacology; D014817:Vitelline Membrane / Q000378:metabolism / Q000648:ultrastructure
D003451:Cryoprotective Agents; D003470:Culture Media; D006536:Heptanes; D006586:Hexanes; D008055:Lipids; D012996:Solutions; C026385:n-hexane; D011092:Polyethylene Glycols; C028618:n-heptane; D014199:Trehalose; D019855:Ethylene Glycol; D009584:Nitrogen; D019840:2-Propanol