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		Materials and Methods The embryos used in this experiment were at the sphere stage of development, which is equivalent to approximately four hours of incubation post-fertilization. More importantly, the sphere stage occurs after the midblastular transition, whereupon the embryo begins to translate its own genes, cell movements occur, and a pattern of slow, asynchronous cleavage begins (Gilbert, 2003). The exposure of each of the variant groups to lithium chloride consisted of a ten-minute incubation period for each so that the time of incubation would remain concurrent with the sphere stage and not extend beyond this period of development. Sphere-stage embryos were isolated according to Stages of Embryonic Development of the Zebrafish (Zebrafish Information Network, 1995). Isolated embryos were collected in Zebrafish Embryo Medium, which was prepared according to the developmental biology lab website (DB Lab, 2003). During the induction perion, approximately 10-15 Brachidanio rerio embryos were treated with the following titrations of LiCl: 0.45 M, 0.30 M, and 0.15 M made up in Zebrafish Embryo Medium. The calculations for the concentrations of Lithium Chloride that were used are as indicated in Table I: Table I: Variable Molarity of Lithium Chloride Titrations Formula weight Experimental concentration (M) Conversion factor Amount LiCl added 42.39 g/ 1 mole 0.45 M/ L .01 L/ 10 mL 0.19 g/ 10 mL 42.39 g/ 1 mole 0.30 M/ L .01 L/ 10 mL 0.13 g/10 mL 42.39 g/ 1 mole 0.15 M/ L .01 L/ 10 mL 0.06 g/ 10 mL The fourth group did not contain any LiCl and served as a control, thus no calculations were required for this titration. Subsequent to the ten-minute incubation period of the three variable groups in the LiCl solutions, all embryos were rinsed with Zebrafish embryo medium and incubated overnight at 28°C.
		J. N. White 5/13/04