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Differentiating effects of murine nerve growth factor in the peripheral and central nervous systems of Xenopus laevis tadpoles.
Levi-Montalcini R
,
Aloe L
.
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The present investigation was directed toward exploration of the spectrum of action of murine nerve growth factor (m-NGF) in peripheral cells and neurons in the central nervous system (CNS) of Xenopus laevis tadpoles. It was found that systemic m-NGF injections elicit growth and differentiative effects not only on sensory and sympathetic nerve cells but also on several populations in the CNS. The finding that aminergic and peptidergic neurons in brain centers are highly receptive to m-NGF activity provides evidence for the broad spectrum of action of this molecule in lower vertebrates and calls for a systematic search for these and other putative target cells in the CNS of higher vertebrates.
FIG. 1. Comparative volume (expressed in arbitrary units) of the
third, fourth, and fifth spinal sensory ganglia of Xenopus tadpoles
injected for 2, 4, and 6 days with saline (c) or NGF (o). Values are
means ± SEM of four determinations.
FIG. 2. (A and B) Comparative sections through the spinal cord
of the third dorsal root ganglia of stage-48 tadpoles treated for 5 days
with vehicle solution (A) and NGF (B). (Toluidine blue stain; x 125.)
(C and D) Comparative sections at the level of the thoracic
paravertebral ganglia in stage-50, vehicle-injected (C) and NGFinjected
(D) tadpoles. The precocious differentiation of sympathetic
ganglia in the NGF specimen (D) is very apparent. (Formaldehydeinduced
fluorescence preparations; x 140.) (E and F) Comparative
sections of the retroaortic area of stage-48 tadpoles treated for 6 days
with vehicle solution (E) and NGF (F). Immature chromaffin cells
are visible in C. In F, the arrow points to a group of sympathetic-like
nerve cells. (Toluidine blue stain; x 185.)
FIG. 3. Mauthner cells in stage-48 tadpoles injected for 6 days
with vehicle (A) and NGF (B). The arrow in B points to well developed
dendritic processes. (Toluidine blue stain; x320.)
FIG. 4. Time-course accumulation of radioactive NGF in the
isolated brain of X. laevis tadpoles at stage 46 injected in the right
periorbital area with 2 A.l of '"I-NGF (e) or 2 1Al of 'MI-IgG (o).
Values are means ± SEM of four determinations.
FIG. 5. Radioautographic histological sections of the CNS of X.
laevis tadpoles at stage 46 following injection with 125I-NGF (50 ng
per tadpole) alone or with an excess of unlabeled NGF; fixation was
2 hr later. (A) Transverse section through the lower spinal cord
showing the dorsal funicular area heavily labeled. (x 140.) (B)
Transverse section through the caudal medulla oblongata showing
the intense labeling of the postotic ganglion, its nerve, and the
afferent dorsal fiber bundles projecting into the medulla. (x 140.) (C
and D) Caudal and rostral brainstem sections showing heavily labeled
fiber tracts afferent to the brainstem from the pre- and postotic
ganglia. In C, intrinsic fiber tracts (arrow) and scattered cell clusters
are also labeled. (C, x 180; D, x 140.) (E) Round-shaped heavily
labeled cells bordering the third ventricle. (x106.) (F) Comparative
histological section injected with 125I-NGF and an excess of
unlabeled NGF. (x106.) (G and H) Sections through rostral
diencephalon injected with 125I-NGF alone (G) and an excess of
unlabeled NGF (H). Intrinsic fiber bundles in the ventrolateral area
are also covered with iodinated NGF. (x73.)
FIG. 6. Formaldehyde-induced fluorescence of tadpole brains at
stage 48 injected for 6 days with vehicle (A, C, E, G) or NGF (B, D,
F, H). (A and B) Comparative brain sections showing aminergic
neurons in the paraventricular organs. (x50.) (C and D) Comparative
brain sections showing the aminergic cells of the nucleus
infundibularis dorsalis. (x 15.) (E and F) Cells ofnuclei infundibularis
dorsalis at higher magnification. (x122.) (G and H) Comparative
brain sections showing fluorescent nerve cells and fiber tracts in the
pontomesencephalic reticular area. (x43.)
FIG. 7. (A and B) SP immunoreactive cells and fiber tracts in the
mesencephalic area of tadpolebrain at stage 48 injected for 6 days
with vehicle (A) or NGF (B). (x86.) (C and D) Som immunoreactive
nerve cells in the pontomesencephalic area of tadpole brains injected
for 6 days with vehicle (C) or NGF (D). (x75.)
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