title: Effects of hypoxia on gene and protein expression in the blue crab, Callinectes sapidus.
abstract: Increases in hypoxic conditions are one of the major factors responsible for declines in estuarine habitat quality, yet to date there are no indicators for recognizing populations of estuarine organisms that are suffering from chronic hypoxic stress. Here we test the hypothesis that alterations in gene and protein expression of antioxidant enzymes and other stress-specific proteins can be used as molecular indicators of hypoxic stress. Blue crabs, Callinectes sapidus, were exposed to 2-3 ppm DO for 5 days. Gene expression was measured using macroarrays constructed from cDNA of 10 partial gene transcripts cloned from blue crab hepatopancreas. Significant (p< or =0.05) down-regulation of gene expression was found for MnSOD, hemocyanin, ribosomal S15 and L23. Subtractive hybridization using RNA from control and hypoxic hepatopancreas tissues also indicated down-regulation of hemocyanin transcription. In contrast, Western blotting showed a significant (p< or =0.05) increase of hemocyanin protein in the hepatopancreas and cross-linking of MnSOD proteins in hypoxia-exposed crabs. Thus, hypoxia-responsive cDNA arrays and Westerns may be useful diagnostic tools for monitoring effects of hypoxia in estuarine crustacea.
authors.affiliation: Department of Coastal Sciences, University of Southern Mississippi, P.O. Box 7000, Ocean Springs, MS 39566, USA.
authors.forename: Marius
authors.initials: M
authors.lastname: Brouwer
authors.forename: Patrick
authors.initials: P
authors.lastname: Larkin
authors.forename: Nancy
authors.initials: N
authors.lastname: Brown-Peterson
authors.forename: Christina
authors.initials: C
authors.lastname: King
authors.forename: Steve
authors.initials: S
authors.lastname: Manning
authors.forename: Nancy
authors.initials: N
authors.lastname: Denslow
mesh_terms: D000199:Actins / Q000235:genetics / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D003386:Brachyura / Q000235:genetics / Q000502:physiology*; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D006433:Hemocyanins / Q000235:genetics / Q000378:metabolism; D043143:Hepatopancreas / Q000378:metabolism; D008668:Metallothionein / Q000235:genetics / Q000378:metabolism; D009693:Nucleic Acid Hybridization / Q000379:methods; D010100:Oxygen / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D012269:Ribosomal Proteins / Q000235:genetics / Q000378:metabolism; D012623:Seawater; D017422:Sequence Analysis, DNA; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism
chemical_list: D000199:Actins; D018076:DNA, Complementary; D018840:HSP70 Heat-Shock Proteins; D012333:RNA, Messenger; D012269:Ribosomal Proteins; D006433:Hemocyanins; D008668:Metallothionein; D013482:Superoxide Dismutase; D010100:Oxygen