Biosynthesis of the Egg-Laying Hormone (ELH) in the Bag Cell Neurons of Aplysia californica

Abstract

Biosynthesis of the egg-laying hormone in the bag cell neurons of Aplysia californica was studied. Bag cells were incubated with leucine-3H in vitro for 30 min and rinsed for variable periods of time in a chase medium. The distribution of incorporated label among proteins within the cells was assayed by electrophoresis of an homogenate on sodium dodecyl sulfate polyacrylamide gels. Results from rinse times shorter than 30 min revealed that the predominant synthetic product is a 25,000 dalton protein. With longer rinse times, this species was reduced and two species of lower molecular weight became prominent. This redistribution of radioactivity was quantitative and was not prevented by inhibition of protein synthesis during the rinse. A 10°C reduction in temperature (from 15°C) blocked the redistribution. These data are interpreted to indicate that the 25,000 dalton molecule is a precursor which is cleaved enzymatically to yield two lower molecular weight products. One product is a 12,000 dalton molecule which remains in the cell bodies. The other is a molecule of <10,000 daltons which is exported from the somata into the neurohemal regions of the connective tissue. Perfusion of these regions with high [K+] medium results in the release of this product into the medium. It is concluded that this product is the 6000 dalton egg-laying hormone (ELH).