Differential expression and regulation by 20-hydroxyecdysone of mosquito ecdysteroid receptor isoforms A and B

Cloning of the AaEcR-A isoform, along with the previously cloned AaEcR-B isoform, has permitted us to evaluate the expression of AaEcR isoforms during mosquito vitellogenesis. Mosquito EcR isoform transcripts exhibited dramatically different patterns of expression after a blood meal-triggered activation of vitellogenesis in the fat body. The AaEcR-B transcript level rose sharply by 4-h post blood meal (PBM), coinciding with the small ecdysteroid peak, and then declined reaching its lowest level at 16-24-h PBM. In contrast, the AaEcR-A transcript peaked at 16-20-h PBM, coinciding with the large ecdysteroid peak. AaEcR-B and AaEcR-A transcripts exhibited a striking difference in sensitivity to 20-hydroxyecdysone (20E), being maximally activated at 10(-8) and 10(-6) M, respectively. Both ecdysteroid receptor (EcR) isoform mRNAs were transcribed in a cycloheximide-independent manner, suggesting that they are direct targets of 20E. However, AaEcR-A transcription requires continuous presence of 20E, while AaEcR-B mRNA level rose for 4 h and then declined under the same conditions. These results indicate the mosquito EcR isoforms play distinct physiological functions during vitellogenesis in the mosquito fat body.