Quantification of juvenile hormone III, vitellogenin, and vitellogenin-mRNA during the oviposition cycle of the lubber grasshopper

The vitellogenic cycle of the lubber grasshopper ( Romalea microptera) was studied by measuring levels of juvenile hormone (JH III), vitellogenin, and vitellogenin-mRNA through the first oviposition cycle. JH III and vitellogenin were measured by radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA), respectively. To measure vitellogenin-mRNA, a partial (753 bp) cDNA fragment of vitellogenin was isolated from the fat body of vitellogenic animals. The sequence of this cDNA was related to vitellogenin sequences in other insect species. Using these sequence data, an RT–PCR (reverse transcriptase polymerase chain reaction) assay was developed to quantify vitellogenin-mRNA levels during the oviposition cycle. Vitellogenin-mRNA levels in the fat body tissue from virgin females were measured on specific days after eclosion and compared to hemolymph levels of JH III and vitellogenin from the same individuals. The levels of all three compounds (JH III, vitellogenin, and vitellogenin-mRNA) showed similar changes throughout the oviposition cycle, being undetectable or nearly undetectable initially (day 3), rising to maximum levels on days 23 and 28, and then dropped to lower or undetectable levels on the day of oviposition. The ability to measure these characteristics will be useful for studying the effects of hormonal and nutritional manipulations on reproduction.