Type of Document Dissertation Author Reynolds, Julie Annette URN etd-10262007-115116 Title Metabolism and Gene Expression during Diapause in Arthropods Degree Doctor of Philosophy (Ph.D.) Department Biological Sciences Advisory Committee
Advisor Name Title Steven C Hand Committee Chair John Lynn Committee Member Joseph Seibenaller Committee Member Mark Batzer Committee Member Michael Stout Dean's Representative Keywords
- Allonemobius socius
- gene expression
- Artemia franciscana
Date of Defense 2007-10-03 Availability unrestricted AbstractArthropods may enter diapause to escape environmental insult. Diapause is an endogenously controlled dormant state defined by developmental arrest and species-specific physiological changes (e.g., metabolic depression and upregulation of compounds that protect cell structure and function). Although physiological changes have been documented for a number of species in diapause, biochemical and molecular regulation of diapause remains largely unexplained.
Aerobic metabolism in diapause, Artemia franciscana, embryos is reduced up to 92 % compared with post-diapause embryos. Differences in isolated mitochondria are insufficient to account for respiratory depression because mitochondria in diapause embryos are structurally similar to mitochondria in post-diapause embryos. Respiratory control ratios and P:O flux ratios of mitochondria from diapause embryos are equal to or higher than those of mitochondria from post-diapause embryos. State 3 and state 4 respiration rates on pyruvate are equivalent in the two stages, and mitochondria isolated from diapause embryos show a moderate, 15-27 % reduction with succinate. Cytochrome c oxidase activity is 53 % lower in diapause embryos, but the minimal impact on mitochondrial respiration appears to be due to the 31 % excess of COX capacity in these embryos.
Allonemobius socius embryos enter diapause 3-4 d post-oviposition as indicated by their morphology and DNA embryo-1. There is not an acute downregulation of metabolism during diapause in this species. Diapause embryos consume O2 at the same rate as morphologically similar non-diapause embryos. Diapause and non-diapause embryos exhibit unusually high [AMP]/[ATP] and low [ATP]/[ADP] during early embryogenesis, suggesting that these embryos may be hypoxic early in development. However, superfusing 3 d embryos with O2 enriched air only partially relieves the hypoxic state, which indicates the unusual energy status is an ontogenetic feature not fully explained by oxygen limitation.
Subtractive hybridization and qPCR identified 6 genes predicted to regulate diapause entry in A. socius. Reptin, TFDp2, CYP450, AKR are significantly upregulated in pre-diapause embryos, and ACLY and Capthesin B-like protease are downregulated compared to non-diapause embryos. The need for genes upregulated in pre-diapause embryos appears to be transient as these genes are substantially downregulated 10 d after diapause entry. Taken together, these studies provide an integrative examination of mechanisms underlying diapause entry in arthropods.
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