Graduate Thesis Or Dissertation
 

Isolation and characterization of the nuclear histones from developing embryos of the brine shrimp, Artemia salina

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  • Techniques were developed for extracting histones from isolated chromatin of encysted embryos of Artemia salina. Histone extracts from embryos collected at eight time intervals after rehydration and initiation of development were analyzed by electrophoresis for qualitative and quantitative changes in composition. Histone to DNA ratios were determined for the same eight times, Proteins were extracted at 17 time intervals from 0 hr up to 60 hr after rehydration. The extracts were subjected to visual analysis by polyacrylamide gel electrophoresis in an effort to determine when new proteins first appear, as an index of differentiation. An attempt was then made to correlate changes in the differentiated state with changes in the nuclear histones. Changes in total protein were not evident until after 16 hr of hydration, when naupliar structures are well-formed. Two changes in histone pattern were observed. The first was the disappearance of a slow-moving band between 0 and 4 hr of development (band 2). The second was the increase in the proportion of the slightly lysine-rich fraction II (band 6) histones between 36 and 48 hr of development, and a decrease in band 3, a lysine-rich histone. The evidence supports the conclusion that genes are activated in order to direct differentiation only several hours after hatching of the nauplius larva has occurred. New proteins which appeared shortly before emergence of the nauplius may be synthesized under the direction of pre-existing mRNA.
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