9602564 Michael Existing technologies will be used in an attempt to resurrect extinct species of Eleutherotlactylus frogs from frozen tissue samples. Previously frozen single sperm will be injected into enucleated eggs of closely related species. Diploid nuclei will be generated from single sperm by pressure inhibition of the first round of cellular cleavage, during which time the genome will replicate once. Initial experiments will be carried out with same-species sperm injections using existing species. Further confirmatory experiments with crossspecies sperm injections using other existing species will be performed before frozen samples from extinct species are utilized. Previously frozen, stored sperm from each of two extinct species will be used with enucleated eggs from two closely related existing species. Resulting frogs will be raised to adulthood and identified as to species by morphological and molecular methods..
