Abstract
Prostaglandins (PGs) and other eicosanoids exert important physiological actions in insects and other invertebrates, including influencing ion transport and mediating cellular immune defense functions. Although these actions are very well documented, we have no information on the mechanisms of PGs actions in insect cells. Here we report on the outcomes of experiments designed to test our hypothesis that PGs modulate gene expression in an insect cell line established from pupal ovarian tissue of the moth Helicoverpa zea (BCIRL-HzAM1 cells). We treated cells with either PGA 1 or PGE 1 for 12 or 24 h then analyzed cell lysates by 2-D electrophoresis. Analysis of the gels by densitometry revealed substantial changes in protein expression in some of the protein spots we analyzed. These spots were processed for mass spectrometric analysis by MALDI TOF/TOF, which yielded in silico protein identities for all 34 spots. The apparent changes in three of the proteins were confirmed by semi-quantative PCR, showing that the changes in mRNA expression were reflected in changes in protein expression. The 34 proteins were sorted into six categories, protein actions, lipid metabolism, signal transduction, protection, cell functions and metabolism. The findings support the hypothesis that one mechanism of PG action in insect cells is the modulation of gene expression.
Highlights
Prostaglandins (PGs) and other eicosanoids are oxygenated, enzymatic metabolites of arachidonic acid and two other C20 polyunsaturated fatty acids
We generated the hypothesis that one mode of PG action in insect cells is their influence on gene expression
The data reported in this paper support our hypothesis that one mechanism of PG action in insect cells is their influence on gene expression as registered by changes in cellular protein profiles
Summary
Prostaglandins (PGs) and other eicosanoids are oxygenated, enzymatic metabolites of arachidonic acid and two other C20 polyunsaturated fatty acids. Eicosanoid structures and biosynthetic pathways are described in several reviews and monographs (Stanley, 2000, 2005, 2006). PGs were first discovered in research into human reproductive physiology (von Euler, 1936) and in the ensuing decades a tremendous amount of information on the biological and biomedical significance of PGs in mammals has accumulated. PGs are present and biologically active in virtually all mammalian body tissues and fluids, where they influence a very wide range of physiological events including ion transport, blood pressure, fever and inflammation.
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