Cytoarchitecture, fiber connections, and ultrastructure of nucleus isthmi in a teleost (Navodon modestus) with a special reference to degenerating isthmic afferents from optic tectum and nucleus pretectalis.

Cytoarchitecture and fiber connections of the nucleus isthmi in a teleost (Navodon modestus) were studied by means of Nissl, Bodian, toluidine blue, Golgi, and Fink-Heimer methods. Synaptic terminals were classified by the ultrastructural characteristics, and their origins were determined by electron microscopic degeneration experiments. The nucleus isthmi is composed of an outer cellular area or shell and an inner noncellular area or core. The shell covers anterior, dorsal, and ventral aspects of the core. The cell bodies in the shell are oval (15 x 20 micrometers) with an anteroposterior long axis, and have many somatic spines. Spines are also seen on the initial segment of the axon. Primary dendrites extend posteromedially and branch out in the core. The core contains thin and thick myelinated fibers, which originate in the optic tectum and in the nucleus pretectalis, respectively. At least two types of axon terminal were distinguished in the nucleus isthmi: S type, containing spherical vesicles, and F type, containing flattened vesicles. S terminals are derived from thin myelinated fibers and are only seen in the core where they form asymmetric synapses with dendrites. Frequently a portion of the S terminal membrane near the usual synaptic cleft is in close apposition with the membrane of an adjacent small dendrite or spine. F terminals, which derived from thick myelinated fibers, make symmetric synaptic contacts with both cell bodies in the shell and dendrites in the core. S terminals degenerate after ipsilateral ablation of the optic tectum, whereas F terminals degenerate after destruction of the nucleus pretectalis.

Fine structure of synapses and retinal innervation of substance P and adenosin deaminase containing neurons in the superior colliculus of the rat.

The fine structure of substance P (SP) and adenosine deaminase (ADA) immunoreactive structures in synaptic contacts localized to the superficial layers of the superior colliculus of the rat was investigated by means of immunoelectron microscopy. We also examined the possibility of retinal innervation of SP- and ADA- containing neurons by immunohistochemistry after degeneration of retinal terminals caused by enucleation. SP-like immunoreactive presynaptic terminals of the stratum griseum superficiale (SGS) formed both asymmetric and symmetric synaptic contacts. Presynaptic dendritelike structures were also observed. SP immunoreactive postsynaptic elements made contacts with terminals showing diverse features. ADA-like immunoreactive structures were seen only as postsynaptic elements to different kinds of nonimmunoreactive terminals and were mostly localized in the ventral third of the SGS and the dorsalmost stratum opticum (SO). After enucleation, degenerating retinal terminals were found to form synaptic contacts with SP and ADA immunoreactive structures. The highest number of such degenerating terminals on ADA immunoreactive structures was observed 2 days after retinal denervation, very few being seen after 5 days. These degenerating terminals were restricted to the ventral SGS and dorsal SO. SP immunoreactive structures postsynaptic to degenerating retinal terminals were most numerous 5 days after enucleation and mainly localized in the dorsal SGS. Occasionally, SP immunoreactive dendritelike processes forming synapses with degenerating retinal terminals were simultaneously presynaptic to other nonimmunoreactive profiles, defining, therefore, serial synapses. The present results suggest that SP-I and ADA-I collicular neurons may be part of distinct channels carrying visual information to the lateral posterior and lateral geniculate nuclei of the thalamus, respectively.

Substance P and enkephalins in the superficial layers of the rat superior colliculus: differential plastic effects of retinal deafferentation.

In this work we studied the effects of unilateral eye enucleation on the contents and distribution of leu-enkephalin-, met-enkephalin-arg6-gly7-leu8-, and substance-P-like immunoreactivities (L-ENK-I, ENK-8-I, and SP-I, respectively) in the superficial layers of the rat superior colliculus (SC) by means of the unlabelled antibody peroxidase-antiperoxidase method. In the normal rat only a few L-ENK-I neurons appear dispersed in the stratum griseum superficiale. No immunostained somata appear in the stratum opticum. The most striking effect of unilateral enucleation was the dramatic appearance of a laminarly distributed population of L-ENK-I and/or ENK-8-I neurons in the dorsal stratum opticum of the SC contralateral to the enucleated side. This population of immunoreactive cells was observed with all the survival times examined in the present study (3, 7, 15, and 30 days) and was always accompanied by an increase in the immunostaining of L-ENK-I and/or ENK-8-I fibers in the contralateral stratum griseum superficiale. Enucleation also produced a decrease in the immunostaining of SP-I dendrites that only became obvious 15 days after enucleation. However, the number of SP-I somata or terminal-like immunoreactive structures showed no detectable changes. These results show that retinal deafferentation of the superficial layers of the rat SC has different effects on some immunohistochemically distinguishable neuronal subgroups in the SC, suggesting different functional or trophic relationships of the retinal input to these groups of neurons.

Laminar and segregated distribution of immunoreactivities for some neuropeptides and adenosine deaminase in the superior colliculus of the rat.

The distribution and morphology of adenosine deaminase, substance P, leucine-enkephalin, corticotropin-releasing factor, and calcitonin gene-related peptidelike immunoreactive cells and fibers throughout the superior colliculus of the rat were examined by means of the unlabelled-antibody peroxidase-antiperoxidase method. Adenosine deaminase immunoreactive cells were found in the stratum opticum and lower stratum griseum superficiale; substance P immunoreactive cells were localized to the upper stratum griseum superficiale, and calcitonin gene-related peptide immunolabelled neurons were situated in deeper strata. Substance P, leucine-enkephalin, and calcitonin gene-related peptide immunoreactive fibers were distributed similarly in their lamination and in their patchlike organization. Corticotropin-releasing factor immunoreactive fibers were observed evenly throughout all the strata and were fewer in the stratum griseum superficiale. These findings suggest that, as in afferent modules and segregated efferents of the mammalian superior colliculus, the cells and fibers containing neuroactive substances and neuroactive substance-related enzymes also show a segregated and laminar distribution.

Effects of eye-enucleation on substance P-immunoreactive fibers of some retinorecipient nuclei of the rat in relation to their origin from the superior colliculus.

We have previously shown that retinal deafferentation causes a decrease in immunoreactive dendrites of substance P-positive neurons of the superficial superior colliculus of the rat. Since some retinorecipient thalamic and pretectal nuclei are putative targets for substance P-containing cells of the superior colliculus, the present study attempted to ascertain whether substance P-immunoreactive fibers in these nuclei are also affected by retinal denervation. We found that unilateral eye removal produced a progressive increase in fibrous substance P immunoreactivity in the nucleus of the optic tract, lateral posterior nucleus, and lateral geniculate nucleus of the side contralateral to the enucleation. On the other hand, unilateral lesions to the superficial layers of the superior colliculus produced a dramatic reduction in substance P immunoreactivity in the ipsilateral nucleus of the optic tract, lateral posterior nucleus, and dorsal and ventral lateral geniculate nuclei. In bilaterally enucleated animals, unilateral lesion to the superior colliculus produced, as expected, loss of immunoreactive fibers only in the lateral posterior nucleus and the retinorecipient nuclei ipsilateral to the lesion. These results suggest that transneuronal changes in the distribution of substance P in collicular neurons observed after enucleation could be reflected in their projections to the other primary visual centers and to the lateral posterior nucleus.

Striatal calcitonin gene-related peptide-like immunoreactive afferents from the regions ventral and medial to the medial geniculate nucleus of rats.

The sources of fibers with immunoreactivity to calcitonin gene-related peptide in the caudal portion of the caudate-putamen were investigated in animals treated by a knife cut, a tracer injection, or an electrolytic lesion of regions ventral and medial to the medial geniculate nucleus. Hemitransection of the brain just caudal to the caudate-putamen decreased ipsilaterally calcitonin gene-related peptide-like immunoreactivity in the caudal portion of the caudate-putamen, suggesting that calcitonin gene-related peptide-like immunoreactive fibers in the caudate-putamen originated from neurons in the area caudal to the caudate-putamen. Some calcitonin gene-related peptide-like immunoreactive neurons in the regions ventral and medial to the medial geniculate nucleus, including the peripeduncular, posterior intralaminar and suprageniculate nuclei, were labeled ipsilaterally with a retrograde tracer after an injection into the caudal portion of the caudate-putamen. In addition, an electrolytic lesion of calcitonin gene-related peptide-like immunoreactive neurons in the regions ventral and medial to the medial geniculate nucleus decreased the number of calcitonin gene-related peptide-like immunoreactive fibers in the ipsilateral caudal portion of the caudate-putamen. These results suggest that some cells containing calcitonin gene-related peptide in several posterior thalamic nuclei project to the caudal caudate-putamen.

Abnormality in the optic nerve of albino mutant quails.

Comparative studies were made between the optic nerves of albino and normal quails. The ipsilateral and contralateral retinofugal projections in the albino quail were similar to those in the normal quail. The abnormality of uncrossed projections in the visual pathways, which is found in albinos of many different mammalian species, was not present in the albino quail. The total cross-sectional area of the optic nerve was approximately 25% smaller in the albino quail than in the normal quail, whereas the fiber density was about 17% higher in the former. The total number of optic nerve fibers in normal and albino quails was therefore the same. Numerous unmyelinated fibers existed in normal and albino quail optic nerves, the ratio of myelinated to unmyelinated fibers being approximately 4:6. Axon diameters of myelinated and unmyelinated fibers in the optic nerves were smaller for the albino quail than for the normal quail. The number of the myelin rings in the albino quail slightly decreased compared with those in the normal quail.

Selective loss of retinal ganglion cells in albino avian glaucoma.

Retinal ganglion cell loss was investigated in the retinae of albino quails before and after the development of glaucoma. The isodensity maps of ganglion cells, the total number of ganglion cells, and the histograms of the cell size in the central region of the retina were similar between albino quails without glaucoma and pigmented quails. However, ganglion cells in the intermediate and peripheral regions of the albino quails retina without glaucoma were significantly smaller than those of the pigmented quail retina. In albino quails with moderate glaucoma in 3 months of age, 11% to 55% of all the retinal ganglion cells had disappeared, with the loss of medium-sized cells (30-60 micron2 occurring earlier than that of small and large cells. In albino quails with advanced glaucoma, there was marked cupping around the optic nerve head, and only small ganglion cells remained in the ganglion cell layer.

Animal model of closed angle glaucoma in albino mutant quails.

The intraocular pressure of imperfect albino mutant quails with a sex-linked recessive gene was measured by using a cannulating method, and morphological changes of the iridocorneal angle were observed with a gonioprism. Albino mutant quails were characterized by an elevated intraocular pressure and lens enlargement at 6 mo after hatching. Initially the iridocorneal angle opened and eventually closed. The usefulness of the closed angle glaucoma as an animal model system for human diseases was discussed.

Increase in participation of vasoactive intestinal peptide in relaxation of the distal colon of Wistar rats with age.

Changes in participation of vasoactive intestinal peptide (VIP) in nonadrenergic noncholinergic (NANC) relaxation of longitudinal muscle of the distal colon with age were studied in 2- to 50-week-old Wistar rats in vitro. The extent of the VIP-mediated component of the relaxation induced by electrical field stimulation (EFS) was determined by the effect of VIP(10 - 28), a VIP receptor antagonist. In 2-week-old rats, the extent of the VIP-mediated component of the relaxation was scarce, about 10%, whereas the component gradually increase with age and reached the maximum extent 66% at 50-week-old. Since our previous results suggest that VIP induces NANC relaxation via activation of charybdotoxin (ChTx, a blocker of large conductance Ca(2+)-activated K(+) channel)-sensitive K(+) channels with concomitant slow hyperpolarization in the muscle cells, we next studied whether ChTx-sensitive component and slow hyperpolarization changes with age. Extent of ChTx-sensitive component of the relaxation increased with age, showing a very similar pattern to VIP-mediated one. EFS induced monophasic inhibitory junction potentials (i.j.ps) in longitudinal muscle cells of the distal colon of 2- and 4-week-old. EFS also induced biphasic i.j.ps in many longitudinal muscle cells of 8- and 50-week-old: rapid and subsequent slow hyperpolarization. A VIP receptor antagonist selectively inhibited the slow hyperpolarization. Exogenously added VIP induced no appreciable change in the membrane potential of longitudinal muscle cells of 2-week-old, whereas it induced slight slow hyperpolarization of the cell membrane in 4-week-old and magnitude of the hyperpolarization increased with age. On the other hand, relaxant response of the longitudinal muscle to exogenously added VIP was high in younger rats. The present results suggest that the role of VIP in mediating NANC relaxation of longitudinal muscle of the Wistar rat distal colon is very little at neonatal stage, but it increases with age.

Geniculo-geniculate projection of enkephalin and neuropeptide Y containing neurons in the intergeniculate leaflet of the thalamus in the rat.

Geniculo-geniculate projections of immunoreactive neurons for gamma-aminobutyric acid (GABA), leucine-enkephalin (ENK), neuropeptide Y (NPY) and substance P (SP) in the intergeniculate leaflet (IGL) and ventral lateral geniculate nucleus (VLG) of the rat were examined by using a combination of retrograde tracing method and immunocytochemistry. After injection of the fast blue (FB) dye into the IGL and VLG, many ENK immunoreactive neurons, and some NPY immunoreactive neurons were labelled by FB dye in the contralateral IGL. However, GABA and SP immunoreactive neurons in the IGL and VLG were not labelled retrogradely by FB dye. A unilateral electrical lesion of the IGL and VLG caused a reduction of the ENK and NPY immunoreactive fibres in the contralateral IGL and VLG. These findings suggested that ENK is one major component of the neuroactive substances in the geniculo-geniculate projection, and NPY also contributes partly to this projection. Furthermore, the unilateral destruction of the IGL and VLG showed a marked loss of the NPY immunoreactive fibres and a slight loss of the ENK immunoreactive fibres in the bilateral suprachiasmatic nuclei.

Localization of neuroactive substances in the rat parabigeminal nucleus: an immunohistochemical study.

The distribution of neurons and fibres that contain substance P, cholecystokinin-8, vasoactive intestinal polypeptide, corticotropin-releasing factor, calcitonin-gene-related peptide, choline acetyltransferase, tyrosine hydroxylase, somatostatin, leucine-enkephalin, and neuropeptide Y was examined in the parabigeminal nucleus of the rat by immunohistochemistry. Many choline acetyltransferase-like immunoreactive or calcitonin-gene-related peptide-like immunoreactive neurons were observed in the dorsal, middle and ventral subdivisions of the parabigeminal nucleus. A few corticotropin-releasing factor-like immunoreactive neurons were also seen in these three subdivisions. The double-immunostaining demonstrated that some choline acetyltransferase-like immunoreactive neurons in the dorsal and ventral subdivisions contained calcitonin-gene-related peptide. Fibres containing cholecystokinin-8, substance P or vasoactive intestinal polypeptide were abundant in the parabigeminal nucleus. Fibres containing cholecystokinin-8 were concentrated in the dorsal and ventral subdivisions, and the lateral margin of the middle subdivision, whereas many fibres containing substance P or vasoactive intestinal polypeptide existed in the lateral half of each subdivision. Fibres containing calcitonin-gene-related peptide or corticotropin-releasing factor were mostly observed around the immunoreactive neurons. Tyrosine hydroxylase-like immunoreactive fibres were scattered in the parabigeminal nucleus.

Differential effects on substance P immunoreactivity in rat suprachiasmatic and olivary pretectal nuclei.

We examined the effects of constant dark or constant light on substance P (SP) and/or neuropeptide Y (NPY) immunoreactive fibres in the rat suprachiasmatic nucleus (SCN) and olivary pretectal nucleus (OPT) by immunohistochemistry. After constant dark, SP immunoreactive fibres and terminals decreased slightly in the SCN, while they increased markedly in the OPT. After constant light, they increased markedly in the SCN, but were little changed in the OPT. NPY immunoreactive fibres and terminals in the SCN decreased slightly after after constant light, but there were no effects on these fibres after constant dark. These findings suggest that SP immunoreactive fibres are involved in mediating illumination discrimination in the SCN and/or OPT.

The organization of the rat lateral geniculate body by immunohistochemical analysis of neuroactive substances.

The organization of neuroactive substances in the rat lateral geniculate body (LGB) was studied with available immunohistochemical stainings. In the dorsal lateral geniculate nucleus (DLG), there existed only gamma-aminobutyric acid (GABA)-like immunoreactive neurons. Immunoreactive fiber plexuses for substance P (SP), cholecystokinin-8 (CCK) and vasoactive intestinal polypeptide (VIP) were present in the lateral margin of the DLG, just beneath the optic tract. There were immunoreactive neurons and fibers for GABA, SP, leucine-enkephalin (ENK) and neuropeptide Y (NPY) in the intergeniculate leaflet (IGL). ENK-, NPY- and SP-like immunoreactive neurons in the IGL were mainly medium-sized, and bipolar or spindle-shaped with a few dendrites oriented dorsoventrally. In the IGL, use of double-labeled immunofluorescence demonstrated that a few neurons exhibited both ENK- and SP-like immunoreactivities, and a few neurons had both GABA- and ENK-like immunoreactivities. Although the morphology of ENK-like immunoreactive neurons resembled to NPY-like immunoreactive neurons, both neurons were clearly different neurons. Many GABA-, ENK- and SP-like immunoreactive neurons and fibers were found in the ventral lateral geniculate nucleus (VLG). These immunoreactive neurons were mainly medium-sized, and bipolar in shape, while a few immunoreactive neurons were of multipolar shape. Neurons containing ENK and fibers containing SP mainly existed in the lateral half of the parvocellular part and in the medial half of magnocellular part of the VLG. In this region, about one-third of the GABA-like immunoreactive neurons contained ENK-like immunoreactivity. Many SP neurons mainly existed in the medial half of the parvocellular part of the VLG. CCK- and VIP-like immunoreactive fibers were present in the lateral half of the magnocellular part of the VLG. Immunoreactive fibers for calcitonin gene-related peptide, corticotropin-releasing factor, neurotensin and tyrosine hydroxylase were disseminated throughout the LGB. The subdivisions of the LGB were discussed, based upon the distribution of neuroactive substances.

Substance P-immunoreactive innervation from the retina to the suprachiasmatic nucleus in the rat.

The present study examined substance P (SP) innervation in the suprachiasmatic nucleus (SCN) of the rat. In the colchicine-untreated rat, SP-immunoreactive fibers formed a dense oval plexus in the ventral part of the SCN. After bilateral eye enucleation, there was a marked reduction in SP-immunoreactive fibers in the ventral part of the SCNs. The SP-immunoreactive neurons in the retinal ganglion cell layer were retrogradely labeled after injection of Fluoro-gold into the SCN. These findings indicate the presence of the SP innervation from the retina to the SCN in the rat. The role of SP in the retino-hypothalamic tract was discussed from the light-dark cycle.

Evidence for the existence of substance P-like immunoreactive neurons in the human cerebral cortex: an immunohistochemical analysis.

Cellular localization of substance P-like immunoreactive (SP-IR) structures in the pre- and postcentral gyri of the human cerebral cortex was examined by indirect immunofluorescence. SP-IR was localized mostly in bipolar and partly in multipolar cells in layers II and IV. SP-IR fibers were also noted in these gyri, especially in layers II and IV.

Ultrastructure and retinal innervation of deafferentation-induced enkephalin-immunoreactive elements in the superficial layers of the rat superior colliculus.

Leu-enkephalin-like immunoreactive (ENK-I) elements appearing in the superficial layers of the rat superior colliculus (SC) after eye-enucleation were examined by means of immunoelectronmicroscopy. ENK-I somata were of a single type and formed symmetric and asymmetric synapses with non-immunoreactive axon terminals. Some degenerating retinal terminals made synaptic contacts only with small ENK-I dendrites, suggesting that deafferentation-induced ENK-I neurons in the rat SC receive retinal input onto the distal portions of their dendrites.

A relationship between substance P receptor and retinal fibers in the rat suprachiasmatic nucleus.

The suprachiasmatic nucleus (SCN) in the hypothalamus controls many of the circadian rhythms in mammalian species. In the present study, we investigated the location of substance P receptor (SPR)-containing neurons in the rat SCN, using a specific antibody against SPR, which corresponds to the NK-1 subtype of tachykinin receptors, and also examined the synaptic relationship between SPR-containing neurons and retinal fibers at the ultrastructural level. An SPR-immunoreactive meshwork of labeled somata and dendrites was identified in the SCN. The strongest SPR-immunoreactivity was observed in the dorsal and lateral parts of the SCN. Many labeled somata were identified there and their dendrites protruded ventrally from their somata. A few SPR-immunoreactive somata were observed also in the ventral part of the SCN and within the optic tract. In the SCN of eye-enucleated animals, degenerating retinal fibers were shown to terminate on SPR-immunoreactive dendrites forming asymmetrical axo-dendritic contacts.

Ipsilateral retinal projections and laminations of the dorsal lateral geniculate nucleus in the eastern chipmunk (Tamias sibiricus asiaticus).

The ipsilateral retinal projections and laminations of the dorsal geniculate nucleus (LGNd) were studied in the eastern chipmunk (Tamias sibiricus asiaticus). From cyto-, myelo- and chemoarchitectures the LGNd was divided into the ventromedial and dorsolateral parts. Anterograde axonal transport of wheat germ agglutinated horseradish peroxidase (WGA-HRP) and Fink-Heimer staining after unilateral eye removal both indicated that the ipsilateral projection terminates in lamina 2 of the ventromedial part and in lamina 3b of the dorsalateral part. The remaining laminas, lamina 1 of the ventromedial part and laminas 3a and 3c of the dorsolateral part, receive afferents from the contralateral retina. It was suggested that laminas 1 and 2 of the chipmunk's LGNd correspond to laminas A and A1 of the cat's LGNd, and the lamina 3 complex to its lamina C complex.

Adenosine deaminase containing fiber pathway from the superior colliculus to the lateral posterior nucleus of the rat.

Presence of a projection containing adenosine deaminase (ADA)-like immunoreactivity from the stratum opticum (SO) to the dorsomedial portion of the lateral posterior nucleus of the thalamus (LPN) of the rat was demonstrated using a method combining retrograde tracing by horseradish peroxidase (HRP) and immunohistochemistry for ADA. In the caudal two thirds of the LPN, a clearly delimited ADA-like immunoreactive fiber plexus was located. Injection of HRP into the lateral posterior nucleus labeled many neurons in the medial portion of the SO where medium-sized neurons with ADA immunoreactivity were concentrated. Simultaneous immunostaining showed that some of the HRP-labeled neurons are ADA-positive.