In vitro activation of insect prothoracic glands by the prothoracicotropic hormone

An in vitro assay for the prothoracicotropic hormone has been developed that utilizes an ecdysone radioimmunoassay to quantify the increase in the rate of ecdysone synthesis elicited by the neurohormonal activation of the prothoracic glands. The rapidity, reproducibility, and accuracy of the assay were maximized by using one member of a gland pair as the control and the other as the test gland. This was possible because the basal rates of ecdysone synthesis by the members of a gland pair were equivalent. Activation was demonstrated to be dose dependent and specific, with prothoracicotropic hormone activity present only in homogenates of brain. The in vitro activation of the prothoracic glands was verified with the Manduca bioassay for the prothoracicotropic hormone in which the morphological responses to the hormone were correlated with increased in vivo ecdysone titers. These results provide unequivocal evidence that the activation of the prothoracic glands by the prothoracicotropic hormone is direct and suggest that activation represents an increase in a basal rate of ecdysone synthesis.