The blood proteins of the cecropia silkworm undergo a complex sequence of changes which are synchronized with the morphological alterations comprising the insect's metamorphosis (15). This relationship indicates that the blood proteins are participants in the biochemical processes which bring about the caterpillar's transformation into the adult moth. In the previous study of this series, six blood proteins were investigated by means of immunological techniques. Each was found to undergo changes in concentration that differed in both magnitude and timing from those of the other five. In order to characterize the cellular mechanisms responsible for these changes, an experimental analysis of the functions of the individual proteins in the metamorphosing insect has been undertaken. The first to be studied in this manner is antigen 7, a female blood protein which occurs only in traces in the blood of males.
Methods
Preparation of AntigenSolutions.raThe antigen solutions were derived from the ceerop/a silkworm (Platysamia cecropia), and included the blood of animals at various stages of metamorphosis, saline extracts of whole adult moths, and the centrifuged yolk of unfertilized eggs. Melanin formation, which normally occurs upon exposure of these solutions to air, was prevented by the addition of a neutralized cyanide solution or a few crystals of phenylthiourea (twice recrystallized from the Eastman product). The collection of blood samples and the preparation of saline extracts of whole animals were performed according to procedures previously described (15).Eggs were obtained from the ovaries of newly emerged female moths, in cecropia as in other Lepidoptera each mature ovary consists of four thin walled tubes (ovarioles) enclosing a linear sequence of up to thirty unfertilized eggs. The ovaries were dissected from anesthetized insects, rinsed several times with saline, and stored at -200C. When subsequently thawed and crushed in a mortar along with a few crystals of phenylthiourea, the eggs yielded a fluid yolk containing large amounts of particulate