Journal of Neuroscience, Vol 6, 2585-2603, Copyright © 1986 by Society for Neuroscience
Tyrosine hydroxylase in the rat parabrachial region: ultrastructural localization and extrinsic sources of immunoreactivity
TA Milner, TH Joh and VM Pickel
We sought to determine the ultrastructural localization and the extrinsic
sources of the catecholamine-synthesizing enzyme, tyrosine hydroxylase
(TH), in the lateral parabrachial region (PBR) of adult male rats. In the
first portion of the study, a rabbit antiserum to TH was
immunocytochemically localized in coronal sections through the lateral PBR
from acrolein-fixed brains using the peroxidase- antiperoxidase method.
Electron-microscopic analysis revealed that perikarya and dendrites with
peroxidase immunoreactivity for TH constituted only 17% of the total
labeled profiles. Afferents to the TH- labeled perikarya and dendrites
usually failed to exhibit immunoreactivity and were thus considered
noncatecholaminergic. Somatic synapses were most commonly detected on small
immunoreactive perikarya in the central lateral nucleus of the PBR. Other
labeled perikarya located in the dorsal lateral or ventral lateral nuclei
received few somatic synapses and were morphologically distinct in terms of
their larger size, infolded nuclear membrane, and abundance of cytoplasmic
organelles. Axons and axon terminals with peroxidase immunoreactivity
constituted the remaining labeled profiles in the lateral PBR. These
terminals primarily formed symmetric synapses with unlabeled and a few
labeled dendrites. The labeled axon terminals were categorized into 2
types: Type I was small (0.3-0.6 micron), contained many small clear
vesicles, and exhibited few well-defined synaptic densities. The second
type was large (0.8-1.4 micron), contained both small clear and large dense
core vesicles, and exhibited well-defined synaptic densities. The 2 types
of terminals were morphologically similar to dopaminergic terminals. The
location of catecholaminergic neurons contributing to the TH-labeled
terminals was determined by combining peroxidase- antiperoxidase
immunocytochemistry for TH with retrograde transport of wheat germ
agglutinin-conjugated horseradish peroxidase (WGA-HRP). The tracer was
unilaterally injected into the PBR of anesthetized adult rats.
Immunocytochemical labeling for TH was seen as a brown reaction product
within neurons in known catecholaminergic cell groups. A black granular
reaction product formed by a cobalt-intensified and
diaminobenzidine-stabilized tetramethyl benzidine reaction for WGA-HRP was
evident within many TH-labeled and unlabeled neurons.(ABSTRACT TRUNCATED AT
400 WORDS)