Yasumitsu Nakai

Localization of phosphoneuroprotein 14 (PNP 14) and its mRNA expression in rat brain determined by immunocytochemistry and in situ hybridization.

Distribution and ultrastructural localization of a novel phosphoneuroprotein with a molecular mass of 14 kDa (PNP 14), and expression of its mRNA were studied in the adult rat central nervous system (CNS) by immunocytochemistry and in situ hybridization histochemistry. PNP 14 immunoreactivity was abundant in the molecular layer, present moderately in the granular layer, and rare in the Purkinje cell layer of the cerebellar cortex. No PNP 14 immunoreactivity was detected in the cerebellar medulla. In the forebrain, immunoreactivity was found in the hippocampus, striatum, and throughout the cerebral cortex, especially in layer V. Electron microscopic immunocytochemical observation in the cerebellar cortex revealed many PNP 14-immunoreactive axon terminals making synaptic contact with dendritic processes in both granular and molecular layers. PNP 14 immunoreactivity was present mainly in the cytoplasmic matrix in the presynaptic axon terminals. PNP 14 mRNA was localized in the granular layer of the cerebellar cortex, in the hippocampus, and in the cerebral cortex, suggesting that PNP 14 is synthesized in neurons of the granular layer and then transported to the molecular layer by axonal transport. These morphological findings suggest that PNP 14 is likely to modulate the function of selected CNS synapses.

Distribution and synaptic relations of NOS neurons in the dorsal raphe nucleus: A comparison to 5-HT neurons

Anti-nitric oxide synthase antibody was used to study the distribution, cytoarchitecture, and synaptic relations of nitric oxide synthase-like immunoreactive neurons in the whole rostral-caudal length of the dorsal raphe nucleus of the rat and compared them with serotonergic neurons. Results showed that the distribution of the nitric oxide synthase in the dorsal raphe nucleus was similar to that of the serotonergic neurons at the rostral part of the dorsal raphe nucleus, including the mediodorsal and the medioventral cell groups, and changed at the middle and caudal parts of the dorsal raphe nucleus. The cytoarchitecture of the nitric oxide synthase-like immunoreactive neurons in the medioventral cell group of the dorsal raphe nucleus was similar to that of the serotonergic neurons. Similar to the serotonergic neurons there, nitric oxide synthase-like immunoreactive neurons also received synapses from axon terminals that contained round, or flattened vesicles, or both kinds. Different to the serotonergic neurons, the few nitric oxide synthase-like immunoreactive axon terminals that were in this area formed synapses.

Direct projections from catecholaminergic neurons in the caudal ventrolateral medulla to vasopressin-containing neurons in the supraoptic nucleus: a triple-labeling electron microscope study in the rat

Direct projections form the A1 catecholaminergic cell group in the caudal ventrolateral medulla (CVLM) to arginine-vasopressin-containing (AVP-containing) neurosecretory neurons in the hypothalamic supraoptic nucleus (SON) were examined electron microscopically in the rat by a triple-labeling technique in which an anterograde tract-tracing method of wheat germ agglutinin-conjugated horseradish peroxidase (WGA-HRP) was combined with immunocytochemistry for tyrosine hydroxylase (TH) and that for AVP. In the SON, TH-like immunoreactive axon terminals which were labeled with WGA-HRP injected in the CVLM were in synaptic contact with neuronal profiles with AVP-like immunoreactivity. The results indicate that AVP-containing neurons in the SON receive monosynaptic catecholaminergic synaptic input from the CVLM.

ELECTRON MICROSCOPIC STUDY OF GABAERGIC SYNAPTIC INNERVATION OF NITRIC OXIDE SYNTHASE IMMUNOREACTIVE NEURONS IN THE DORSAL RAPHE NUCLEUS IN THE RAT

A double immunocytochemical method combining the preembedding avidin biotin peroxidase complex technique and the postembedding immunogold technique was used to examine synaptic interactions between GABAergic and nitric oxide synthase containing neurons in the same tissue sections of the dorsal raphe nucleus of the Wistar white rat. Although a large number of immunogold stained GABAergic axon terminals were found to be presynaptic to dendrites containing nitric oxide synthase‐like immunoreaction product, synapses between GABA‐like immunoreactive axon terminals and nitric oxide synthase‐like immunoreactive perikarya were rare. The labeled boutons were found to make symmetrical and asymmetrical synapses. No axo‐axonic synapse was found. These results suggest that GABAergic neurons could modulate nitric oxide producing neurons in the dorsal raphe nucleus through direct synaptic relations. Synapse 25:24–29, 1997.

Immunoelectron microscopy of enkephalinergic innervation of GABAergic neurons in the periaqueductal gray

The pre-embedding double immunoreaction method was used to study synaptic relations of enkephalinergic and GABAergic neuronal elements in the ventrolateral part of the periaqueductal gray of the Wistar albino rat. The enkephalin-like neuronal elements were immunoreacted by the silver-gold intensified peroxidase-antiperoxidase method and the GABA-like immunoreactive neurons were immunoreacted by the unintensified peroxidase-antiperoxidase method. GABA-like immunoreactive neuronal somata were post-synaptic to both the enkephalin-like immunoreactive and the non-immunoreactive axon terminals. Enkephalin-like immunoreactive axon terminals were found to make synapses with GABA-like immunoreactive and non-immunoreactive dendrites. The synapses between the two kinds of chemically characterized neurons appeared to be both asymmetrical and symmetrical. Possible functional activity related to pain modulation, and synaptic relations between the enkephalinergic and GABAergic neurons in the periaqueductal gray and the dorsal raphe nucleus, are discussed.

Immunoelectron microscopy of β-endorphinergic synaptic innervation of nitric oxide synthase immunoreactive neurons in the dorsal raphe nucleus

On the basis of the comparing of the distribution of β-endorphin-like immunoreactive neuronal fibers and nitric oxide synthase-like immunoreactive neurons in the dorsal raphe nucleus, the synapses between the two immunocytochemically identified neurons were studied with a modified DAB-silver-gold intensification double immunostaining technique at the electron microscopic level. Although both of them can be found in the mediodorsal and medioventral parts of the dorsal raphe nucleus, the synapses between them could only be found in the mediodorsal part. The majority of the β-endorphin-like immunoreactive neuronal fibers contained many dense-cored vesicles. The synapses made by β-endorphin-like immunoreactive neuronal axon terminals on nitric oxide synthase-like immunoreactive neurons were both symmetrical and asymmetrical with the former predominant, especially in the axo-dendritic ones. β-Endorphin-like immunoreactive perikarya could only be found in the ventrobasal hypothalamus. These findings suggest the possibility that the β-endorphin- producing neurons in the ventrobasal hypothalamus could influence nitric oxide synthase-containing neurons in the dorsal raphe nucleus by synaptic relations.

Electron microscopic study of GABAergic synaptic innervation of neurotensin-immunoreactive neurons in the dorsal raphe nucleus

A double immunocytochemical method combining the preembedding avidin-biotin-peroxidase-complex technique and the postembedding immunogold technique was used to examine synaptic interactions between GABAergic and neurotensin-containing neurons in the same tissue sections of the dorsal raphe nucleus of the rat. Whereas the neurotensin-like immunoreactive perikarya rarely received synapses from GABA-like immunostaining axon terminals, the neurotensin-like immunoreactive dendrites frequently received synapses from GABA-like immunoreactive neurons. These results suggest that GABAergic neurons could modulate neurotensinergic neurons in the dorsal raphe nucleus through synaptic relations. The immunocytochemically identified local synaptic circuit in the dorsal raphe was discussed.

Synaptic relations of neurotensinergic neurons in the dorsal raphe nucleus

The ultrastructure and synaptic relations of neurotensinergic neurons in the rat dorsal raphe nucleus (DRN) were examined. The neurotensin-like immunoreactive (NT-L1) neurons in the DRN were fusiform or spherical. The NT-LI perikarya could only be detected in colchicine-treated animals whereas the immunoreactive axon terminals could only be found in the animals not treated with colchicine. Although many NT-LI dendrites received synapses from nonimmunoreactive axon terminals, the NT-LI perikarya received few synapses. NT-LI axon terminals also made synapses on nonimmunoreactive dendrites. Occasionally, synapses were found between the NT-LI axon terminals and NT-LI dendrites in the cases in which the animals were not treated with colchicine.

Reciprocal synaptic relations between enkephalinergic and GABAergic neurons in the area postrema of the rat

A preembedding, double immunostaining technique was used to study synaptic relations between enkephalinergic and GABAergic neurons in the area postrema of the rat. As a main result, the enkephalinergic dendrites received many synapses from GABAergic axon terminals, and most of the synapses were symmetrical. Enkephalinergic neuronal perikarya received a few synapses from GABAergic axon terminals, and a few enkephalinergic axon terminals were found presynaptic to GABAergic neurons. Synapses between enkephalinergic profiles were frequent, but no axo-axonic synapses were seen. These findings suggest that GABAergic innervation of enkephalinergic neurons is the main relation between the two kinds of neurons in the area postrema. The synapses between the enkephalinergic axon terminals and GABAergic neurons might be explained as being part of the local servo system of the area postrema.

The reciprocal synaptic relations between enkephalinergic neurons and catecholaminergic neurons in the area postrema

A preembedding double immunostaining technique using antibodies against methionine-enkephalin and tyrosine hydroxylase was used to study synaptic relations between enkephalinergic and catecholaminergic neurons in the area postrema of the rat at the electron microscopic level. The large nuclei-containing cell bodies of the catecholaminergic neurons displayed well-developed Golgi apparatus. The catecholaminergic somata and dendrites received synapses from enkephalinergic axon terminals, and most of the synapses were symmetrical. Occasionally, the catecholaminergic axon terminals were also found to be presynaptic to the enkephalinergic dendrites. Because the enkephalinergic neurons have been reported to be involved in cardiovascular function and the catecholaminergic neurons involved in the vomiting behavior, the synapses observed in this study may provide morphological evidence of the relationship between the vomiting and cardiovascular functions that are triggered in the area postrema.