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Journal of Neuroscience, Vol 9, 4122-4137, Copyright © 1989 by Society for Neuroscience
Hypothalamic expression of a novel gene product, VGF: immunocytochemical analysis
AN van den Pol, C Decavel, A Levi and B Paterson
Section of Neurosurgery, Yale Medical School, New Haven, Connecticut 06510.
VGF is the designation for a new 712 amino acid protein, regulated by nerve
growth factor (NGF) in PC12 cells, that has not been previously described
in the CNS. Northern blot analysis with a nick-translated VGF cDNA probe
revealed a single band of mRNA in the brain with a molecular weight
identical to that found in PC12 cells. The current paper presents a series
of immunocytochemical studies of VGF expression with a focus on the
hypothalamus. Two different antisera were raised against nonoverlapping
amino acid sequences of a bacterial-expressed protein from the VGF gene
cloned from PC12 cells. VGF immunoreactivity is strongly expressed in the
rat suprachiasmatic nucleus (SCN), particularly in the dorsomedial part of
the nucleus. The administration of colchicine to block axonal transport
facilitates detection of the VGF immunoreactivity also in the ventrolateral
suprachiasmatic nucleus. This protein appears to be the first one of
limited neuronal distribution which is found in both dorsomedial SCN and
ventrolateral SCN. Immunostaining of serial 1 micron SCN sections reveals
co- localization of VGF in cells which also contain vasopressin or
vasoactive intestinal polypeptide. Weaker immunoreactivity is also found in
the magnocellular paraventricular and supraoptic nuclei, where the VGF
immunoreactivity co-localizes with oxytocin or vasopressin. Mutant
Brattleboro rats which do not express vasopressin showed strong VGF
immunoreactivity both in the dorsomedial SCN and in cells of the
magnocellular neuronal systems, including cells which normally express
vasopressin. When axonal transport of the protein is blocked by colchicine,
VGF-immunoreactive cells in the hypothalamic arcuate, parvocellular
paraventricular, and tuberomammillary nuclei can also be detected, in
addition to weakly immunoreactive scattered cells in the hippocampus,
amygdala, thalamus, and cortex. VGF immunoreactivity is strong in the
axonal projections of SCN and weak in the axons of the paraventricular and
supraoptic nuclei. With ultrastructural studies, VGF immunoreactivity is
found in presynaptic boutons in the SCN and in axons in the
neurohypophysis. Weak axonal staining is present in some regions of the
hypothalamus and in the external and internal zones of the median eminence.
Immunoreactivity is absent from the intermediate lobe of the hypophysis. In
neonatal rats strong VGF immunoreactivity is found throughout the SCN at
postnatal day 4 but not in the adjacent hypothalamus. VGF immunoreactivity
is also seen in other areas of the brain in neonatal rats, including the
lateral geniculate nucleus; while the staining in the dorsal lateral
geniculate disappears in the adult, that in the intergeniculate leaflet, a
visual center which projects to the SCN, remains.(ABSTRACT TRUNCATED AT 400
WORDS)
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