Cytochrome P-450 monooxygenases in the mosquito, Culex pipiens Public Deposited

http://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/xw42nb959

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  • A sucrose density-gradient ultracentrifugation technique for obtaining microsomes has been developed. This technique has advantages over the classic differential centrifugation technique, in that it is faster, is better suited to work with small quantities of tissue, and offers the opportunity to work with a defined microsomal sub-cellular fraction, rather than a pellet assumed to contain the desired cellular organelles. This procedure has been applied in conjunction with others to study changes in microsomal monooxygenase activities in adult female mosquitoes (Culex pipiens L.) of different ages and reproductive states. Evidence has been obtained that microsomal aldrin epoxidation is a cytochrome P-450 dependent activity in this species. Changes in microsomal aldrin epoxidation and NADPH-cytochrome c reductase activities in mosquitoes of different ages, both before and after a blood meal , have been determined and correlated with changes in whole-body ecdysteroid titers and growth of follicles. The periods of maximal enzymatic activity (second day after emergence and 48 to 96 hours after the blood meal) follow the peaks in ecdysteroid titers by 12 to 24 hours and coincide with the periods of maximal follicle growth and the onset of oviposition. Ecdysteroids were detectable by radioimmunoassay in non-blood-fed mosquitoes on the first day after emergence (15.1 fmol 20-hydroxy-ecdysone equivalents per insect). Ecdysteroids rose to a peak (394 fmol 20-hydroxyecdysone equivalents per insect) at 36 hours after the blood meal. A critical period for release of a head factor which controls vitellogenic development of the follicles was determined by decapitation experiments to be 4 to 8 minutes after beginning but before completing the blood meal.
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