|aDepartment of Chemistry & Biochemistry, BioSciences West Room 518, 1041 E. Lowell St., University of Arizona, Tucson, AZ 85721, United States|
Amy Alabastera, Jun Isoea, Guoli Zhoua, Ada Leea, Ashleigh Murphya, W. Anthony Dayb, Roger L. Miesfelda,c
c Center for Insect Science, BioSciences West Room 518, 1041 E. Lowell St., University of Arizona, Tucson, AZ 85721, United States
Received 21 July 2011; revised 3 September 2011; Accepted 21 September 2011. Available online 29 September 2011.
To better understand the mechanism of de novo lipid biosynthesis in blood fed Aedes aegypti14C-leucine as a metabolic precursor of 14C-acetyl-CoA, we found that 14C-triacylglycerol and 14C-phospholipid levels were significantly reduced in both ACC and FAS1 deficient mosquitoes, confirming that ACC and FAS1 are required for de novo lipid biosynthesis after blood feeding. Surprisingly however, we also found that ACC deficient mosquitoes, but not FAS1 deficient mosquitoes, produced defective oocytes, which lacked an intact eggshell and gave rise to inviable eggs. This severe phenotype was restricted to the 1st gonotrophic cycle, suggesting that the eggshell defect was due to ACC deficiencies in the follicular epithelial cells, which are replaced after each gonotrophic cycle. Consistent with lower amounts of de novo lipid biosynthesis, both ACC and FAS1 deficient mosquitoes produced significantly fewer eggs than control mosquitoes in both the 1st and 2nd gonotrophic cycles. Lastly, FAS1 deficient mosquitoes, but not ACC deficient mosquitoes, showed delayed blood meal digestion, suggesting that a feedback control mechanism may coordinate rates of fat body lipid biosynthesis and midgut digestion during feeding. We propose that decreased ACC and FAS1 enzyme levels lead to reduced lipid biosynthesis and lower fecundity, whereas altered levels of the regulatory metabolites acetyl-CoA and malonyl-CoA account for the observed defects in eggshell formation and blood meal digestion, respectively. mosquitoes, we quantitated acetyl-CoA carboxylase (ACC) and fatty acid synthase 1 (FAS1) transcript levels in blood fed mosquitoes, and used RNAi methods to generate ACC and FAS1 deficient mosquitoes. Using the ketogenic amino acid
The Chicken anti-Bovine Serum Albumin used in this study is available from Gallus Immunotech.