In vitro biosynthesis of juvenile hormone by the larval corpora allata of Manduca sexta: quantificationby radioimmunoassay.

An in vitro method has been developed for the investigation of the regulation of juvenile hormone biosynthesis by insect corpora allata. Glands were maintained in Marks medium 19AB and JH synthesis quantified by a modified radioimmunoassay for juvenile hormone I. The radioimmunoassay is specific for JH I and exhibits approximately 12.6% cross reactivity with JH II and no cross reactivity with JH III. The assay directly measures the JH present in culture medium and has a maximum sensitivity of 50 pg JH I equivalents. Corpora allata from day 5 last instar Manduca sexta larvae were used to define the kinetics parameters of the in vitro system, including a demonstration that small groups of right and left glands synthesize equivalent amounts of juvenile hormone. The juvenile hormones synthesized were identified as juvenile hormones I and II in a ratio of 1:4, respectively. Juvenile hormone III could not be excluded as a product of the corpora allata owing to the low cross reactivity of this homolog (1.7%) in the radioimmunoassay. Corpora allata from different developmental stages exhibited synthesis rates generally consistent with predicted activity based on in vivo hormone titers with the exception of day 5 of the last instar. The variation in gland activity relative to the control of juvenile hormone titer in vivo is discussed.