CYTODIFFERENTIATION IN THE ACCESSORY-GLANDS OF TENEBRIO-MOLITOR .8. CROSSED IMMUNOELECTROPHORETIC ANALYSIS OF TERMINAL DIFFERENTIATION IN THE POSTECDYSIAL TUBULAR ACCESSORY-GLANDS

The terminal differentiation of the tubular accessory glands in the postecdysial male mealworm beetle, T. molitor, is characterized by the synthesis of 4 major groups of proteins as analyzed by 1-dimensional SDS[sodium dodecyl sulfate]-polyacrylamide gel electrophoresis. On 2-dimensional gels (SDS and pI [isoelectric point]), 2 of these groups of proteins (A and B) are acidic in nature and have apparent MW of 17,500 and 18,800, respectively. A 3rd protein group (C) is heterogeneous with respect to isoelectric point (neutral to basic) and has a MW of .apprx. 21,900. The 4th class of proteins (D) is divisible into 2 subgroups on the basis of isoelectric points and has MW ranging from 26,400-29,100. Antibodies were produced to the soluble portion of mature tubular accessory glands. The preparations of heterologous antisera were shown to be gland specific following absorption with bean-shaped accessory gland homogenates and ammonium sulfate fractionation. The 1st appearance and subsequent accumulation during postecdysial maturation of some of these proteins were determined by crossed immunoelectrophoresis. The A and B proteins are detected immunologically, initially in the late pupa, and their quantities remain unchanged until shortly after adult eclosion. During the 8 days following adult eclosion each of these proteins increases in quantity from .apprx. 10-7000 ng. A 3rd immunoreactive protein appears in the 2-day old adult and increases to .apprx. 960 ng over the following 6 days. The identity of this antigen is uncertain although it may represent either the C or the D protein. Based on crossed immunoelectrophoresis, it is evident that at least 2 of these tubular accessory gland antigens contribute to the spermatophore.