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Biomedical subjects

K Shinoda

Publications and source records attributed to K Shinoda.

At least 145 records · Page 8Linked to original sources
PubMed1988

Interaction between adrenergic fibers and intermediate cholinergic neurons in the rat spinal cord: a new double-immunostaining method for correlated light and electron microscopic observations.

Relationships between cholinergic neurons and adrenergic fibers in the intermediate region of the rat thoracic spinal cord were examined using a new immunohistochemical double-staining method for light and electron microscopic observations. Cholinergic neurons were labeled by a monoclonal antibody to choline acetyltransferase and stained bluish green by 5-bromo-4-chloro-3-indolyl-beta-D-galactoside reaction products using beta-galactosidase as a marker. On the same sections, adrenergic fibers were labeled by a polyclonal antiserum to phenyl-ethanolamine-N-methyltransferase and stained brown by diaminobenzidine reaction products using peroxidase as a marker. After embedding in Epon, the sections were examined in the light and electron microscopes. In the light microscope, choline acetyltransferase-like immunoreactive cells were seen in the four discrete areas of the intermediate region: the principal intermediolateral nucleus, the central autonomic nucleus, the intercalated nucleus and the funicular intermediolateral nucleus. These cell groups seemed to be connected to each other by their processes, and they showed a "ladder-like appearance" as a whole. Phenylethanolamine-N-methyltransferase-like immunoreactive fibers were present only along this "ladder-like structure" and were the most rich in the principal intermediolateral nucleus. In the electron microscope, some of the choline acetyltransferase-like immunoreactive neurons, which were identified by light micrographs, were found to receive synaptic inputs from phenylethanolamine-N-methyltransferase-like immunoreactive boutons in the principal intermediolateral nucleus. These findings suggest that the adrenergic axons in the principal intermediolateral nucleus directly affect the activity of the cholinergic preganglionic sympathetic neurons.

Adrenergic Fibers↗
PubMed1988

A reliable method combining horseradish peroxidase histochemistry with immuno-beta-galactosidase staining.

A sensitive combination of horseradish peroxidase (HRP) tracing and immunohistochemistry was used by Rye et al. [J Histochem Cytochem (1984) 32:1145] in a search for the origins of neurotransmitter- and neuromodulator-containing nerve fibers in brain. In this combination, peroxidase as a marker in immunohistochemistry was thought to yield a homogeneous brown immunoreaction product of diaminobenzidine, different from the black granular reaction product of retrogradely transported HRP, which is visualized by the tetramethylbenzidine (TMB) reaction and subsequent stabilization. A neuron that exhibits both kinds of reaction products in its cytoplasm in sections subjected to combination staining is referred to as a double-labeled cell. With a combined HRP and corticotropin-releasing factor (CRF) immunoperoxidase-antiperoxidase (PAP) method, the first set of experiments showed "false" double-labeled cells in the pyramidal cell layer of rat cerebral cortex, but only rarely in the subcortical areas, possibly because of the use of one enzyme system in two different histochemical procedures. This limitation of the double-staining technique prompted us to demonstrate an alternate combination of HRP tracing and immunohistochemistry in the second set of experiments by employing two previously described independent enzyme systems: HRP as a retrograde tracer and beta-galactosidase as a marker for immunohistochemical demonstration of CRF. A homogeneous blue reaction product indicated immuno-beta-galactosidase staining, and a granular black or brown reaction product labeled retrogradely transported HRP in double-labeled cells in subcortical regions. Neither double labeling nor "false" double labeling was seen in pyramidal cells of cerebral cortex. These findings suggest that application of two independent enzyme systems in a combined HRP and immunohistochemical method may be useful for investigating in origins of peptidergic fibers in brain when the combination of HRP histochemistry and the PAP method appears to be inappropriate.

Animals↗
PubMed1987

The histaminergic innervation of the mesencephalic nucleus of the trigeminal nerve in rat brain: a light and electron microscopical study.

Histaminergic fibers in the mesencephalic nucleus of the trigeminal nerve of Long-Evans rats were examined by light and electron microscopy after peroxidase-antiperoxidase immunocytochemical staining for histidine decarboxylase (HDC) as a marker. By light microscopy, neurons in the mesencephalic nucleus of the trigeminal nerve were seen to be surrounded by a number of HDC-like immunoreactive (HDCI) fibers, suggesting the presence of axo-somatic contact. This finding was supported by immunoelectron microscopic demonstration of synaptic contact of some HDCI fibers with the soma of the neurons in this nucleus. These findings indicate that histamine is involved in the sensory regulation of movement of the masticatory muscles at the level of the trigeminal mesencephalic nucleus.

Animals↗
PubMed1987

Hippocampofugal gamma-aminobutyric acid (GABA)-containing neuron system in the rat: a study using a double-labeling method that combines retrograde tracing and immunocytochemistry.

Using a double-labeling method that combines retrograde tracing and immunocytochemistry, we demonstrated the presence of a hippocampofugal gamma-aminobutyric acid (GABA)-like immunoreactive (GABA-LI) neuron system in the rat. As a preliminary experiment, knife-cut of the dorsal fornix was made and it resulted in a remarkable accumulation of GABA-LI fibers caudal to the section. This suggested the presence of a hippocampofugal GABAergic projection. The hypothesis was confirmed by the double-labeling method: injection of the biotinylated wheat germ agglutinin (B-WGA) into the septum resulted in labeling numerous neurons in the hippocampus and some of them also showed GABA-like immunoreactivity by the simultaneous immunostaining with the antiserum against GABA.

Animals↗
PubMed1987

Analysis of the habenulopetal enkephalinergic system in the rat brain: an immunohistochemical study.

The enkephalinergic afferent system in the rat habenula was examined by the indirect immunofluorescence method with antibodies against leucine-enkephalin. Leucine-enkephalin-like immunoreactive (L-ENKI) fibers were observed in the dorsal portion of the medial habenular nucleus (MHb), the intermediate portion of the lateral habenular nucleus (LHb), and the border zone between the MHb and the LHb (BZHb). Knife-cut studies demonstrated that almost all the fibers were supplied via the stria medullaris. Also two discrete ENKergic afferents to the MHb and the LHb were found by several kinds of lesion studies: the MHb was ipsilaterally innervated from L-ENKI neurons in "septoperiventricular area," which is the junctional area between the ventral supracommissural septum and the rostral thalamic periventricular region. The LHb was ipsilaterally innervated from L-ENKI neurons in the rostral portion of the lateral hypothalamus. Our findings also suggested that the L-ENKI fibers in the BZHb are supplied via the stria medullaris with an ipsilateral predominance and that, at the most caudal level, they arise not only from the ipsilateral stria medullaris but also from the contralateral stria medullaris via the habenular commissure.

Animals↗