Journal of Neuroscience, Vol 4, 1840-1849, Copyright © 1984 by Society for Neuroscience
Biosynthesis and axoplasmic transport of neurophysins in the hypothalamo-neurohypophysial system of Rana pipiens
AC Chang and HD Dellmann
Biosynthesis, axoplasmic transport, and storage of neurophysin in the
amphibian (Rana pipiens) magnocellular peptidergic neurosecretory system
were studied, and the results were compared with those reported in mammals.
After injection of [35S]cysteine into the preoptic recess, light
microscopic autoradiography provides evidence that neurons of the preoptic
nucleus (PON) synthesize cysteine-rich proteins. The time course of
appearance of these [35S]cysteine-labeled proteins in different regions of
the hypothalamo-neurohypophysial system was studied by slab gel
autoradiography. [35S]Cysteine-labeled proteins were found in the PON less
than 1 hr postinjection, whereas a major labeled protein, tentatively
identified as the neurophysin, first appeared in the infundibulum and
neural lobe 4 hr after the injection. In addition, the labeled neurophysin
persisted in the neural lobe throughout the entire observation period of 5
days. The minimum transport rate for neurophysin was calculated as 0.9
mm/hr (22 mm/day) at 25 degrees C. Two different neurophysins (with
isoelectric points (pI) 4.9 +/- 0.1, 4.6 +/- 0.1, and Mr = 23,000, 20,100)
may be resolved from the neural lobe extracts by isoelectric focusing and
SDS- polyacrylamide gel electrophoresis, respectively. In addition to the
neurophysin peaks, two radioactive peaks with pI 5.2 and 5.8 may be
detected in the preoptic nucleus and the infundibulum as early as 30 min
after [35S]cysteine injection. Preliminary conversion studies suggest a
putative precursor role for the pI 5.2 protein. The results indicate that
in the amphibian peptidergic neurosecretory system, the synthesis of
cysteine-rich neurophysin by the preoptic neurons, the transport through
the infundibulum, and the storage in the neural lobe proceed similarily to
their mammalian counterparts.