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The primary olfactory projections of channel catfish Ictalurus punctatus have been examined with postmortem tracing by using either 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate or 1,1-dilinoleyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI). Following DiI deposition into small areas in different parts of the posterior olfactory bulb, olfactory sensory neurons always were labeled throughout the olfactory epithelium. No obvious topographical mapping exists between the epithelium and olfactory bulb. The different dye placements, however, did result in labeling of different morphologies of receptor cells, depending on the site of injection. Retrogradely labeled neurons in the olfactory epithelium were classified into three types on the basis of their height: tall, intermediate, and short. Tall olfactory sensory neurons had perikarya at the bottom one-fourth of the epithelium, extended slender dendrites to the epithelial surface, and possessed numerous cilia on the apical dendritic tips. These tall olfactory sensory neurons were labeled predominantly following DiI applications to the ventral part of the posterior olfactory bulb. In contrast, the short olfactory sensory neurons had perikarya situated within the superficial half of the epithelium and with short apical dendrites bearing microvilli. These short olfactory sensory neurons projected predominantly to the dorsal, posterior olfactory bulb. Thus, short microvillous receptor cells and tall ciliated receptor cells connect to different parts of the olfactory bulb, although the receptor cells are intermingled within the olfactory epithelium. Because different parts of the olfactory bulb are thought to respond preferentially to different classes of odorants, these results suggest that receptor cell morphology may be related to odorant quality detection. In addition, to compare this study with previous in vivo studies, Fluoro-Gold was injected in vivo into either the olfactory bulb or intraperitoneally. These in vivo studies show that so-called "type II ciliar receptor cells" of the nonsensory epithelium are labeled nonselectively by blood-borne substances, but they are not labeled by postmortem injections of DiI anywhere in the olfactory bulb.

The calcium ion plays an important role in some critical developmental events in the nervous system, such as neurulation and neurite elongation. Therefore, as the intracellular calcium-binding proteins calbindin-D28k (CaB) and parvalbumin (PV) may be expressed in these developmental events. Accordingly, the ontological expression of CaB and PV was examined immunocytochemically in the spinal cord and dorsal root ganglia (DRG) of the rat, in order to evaluate the relationship between CaB and PV expression, and other important developmental events. During the ontogenesis of the spinal cord, the CaB-like immunoreactivity was mainly observed in the cell somata. The immunoreactive cells in the ventral horn of the cervical and thoracic, lumbar, and sacral segments first appeared at embryonic day (E)-12, E-13, and E-14, respectively. However, these cells were not detected in the intermediate gray matter of the same segments at E-14, E-15, and E-16, respectively, and in the dorsal horn at E-14-E-15, E-16, and E-17, respectively. The peak of immunoreactive cells, both as to number and intensity, occurred in the perinatal period. However, from postnatal day (P)-14 on, the number and intensity of the positive cells decreased, the adult levels being reached at P-35. The PV-like immunoreactivity was mainly detected in the fibers and punctata during the ontogenesis of the spinal cord. The immunoreactive fibers first appeared on the surface of the dorsal horn in the cervical and thoracic segments at E-14, then entered the dorsal horn at E-15, and reached the intermediate gray matter and ventral horn at E-16. The first appearance of these fibers in the same areas of the lumbar and sacral segments occurred 1 day later than in the cervical and thoracic segments. During the perinatal period, the maximum content of PV-like immunoreactive fibers, together with many punctata, was seen in the gray matter. However, between P-14 and P-17, most of them lost immunoreactivity rapidly, with the exception of the medial region of the intermediate gray matter, where the PV-immunoreactive punctata remained up to the adult stage. In DRG neurons, both CaB and PV was expressed, but in different neurons. Neurons labeled with anti-CaB and anti-PV sera were first detected at E-16 and E-14, respectively. These neurons were large or medium-sized in the prenatal period.(ABSTRACT TRUNCATED AT 400 WORDS)

Postnatal developmental changes of preproenkephalin (PPE) gene expression in rat brainstem neurons were studied by in situ hybridization histochemistry. On the basis of PPE mRNA expression, brainstem neurons were categorized into three types: 1) type I neurons were characterized by constant or increasing expression of PPE mRNA during postnatal development; 2) type II neurons started to express PPE mRNA several days after birth and continued to do so thereafter; and 3) type III neurons showed transient expression of PPE mRNA or stopped expressing the mRNA during early postnatal development. Type I PPE neurons were observed in diverse brainstem structures including the mesencephalic and pontine central gray matter, various reticular and raphe nuclei, the ventral tegmental area of Tsai, the interpeduncular nucleus, the nucleus of the brachium of the inferior colliculus, the ventral and dorsal tegmental nuclei of Gudden, the sphenoid nucleus, the laterodorsal tegmental nucleus, Barrington's nucleus, the parabrachial region, the lateral lemniscus and its related nuclei, the trapezoid nucleus, the rostral and ventromedial periolivary nuclei, the mesencephalic trigeminal and principal sensory trigeminal nuclei, the locus coeruleus, the subcoeruleus nucleus, the medial and spinal vestibular nuclei, the dorsal and ventral cochlear nuclei, the medial and lateral cerebellar nuclei, the Roller nucleus, and the intermedius nucleus of the medulla. Type II PPE neurons were found in the superior colliculus, the inferior colliculus, the central part of the dorsal tegmental nucleus, and as Golgi neurons in the granular layer of the cerebellum. Type III PPE neurons were located in the substantia nigra, the red nucleus, the superior olive, the motor trigeminal nucleus, the facial nucleus, the inferior olive, the dorsal motor nucleus of the vagus, and the hypoglossal nucleus. Such region-specific expression of the PPE gene during postnatal ontogeny suggests that rat brainstem PPE neurons may be involved in a variety of developmental events, such as cell proliferation, differentiation, and migration.

The central fiber connections of the gustatory system (VII, IX, and X nerves) in the crucian carp were examined by the Fink-Heimer method and its modification. The sensory and recurrence roots of the VII enter the brainstem separately and terminate in the ipsilateral half of the facial lobe (L-VII). Afferent fibers of the IX terminate in the glossopharyngeal lobe (L-IX). Most afferent fibers of the X terminate in the sensory layer of the vagal lobe (L-X), in which degenerating terminals occur in some laminae. Some vagal afferents project bilaterally to the commissural nucleus of Cajal. The cutaneous component of the X projects to the nucleus of the spinal trigeminal tract (SpV) and the medial funicular nucleus (nFM). Ascending secondary fibers from the L-VII project bilaterally to the secondary gustatory nucleus (nGS) in the isthmus region. Descending secondary fibers from the L-VII turn caudally in the SpV. These fibers terminate mostly in the nucleus of the SpV and sparsely in the nFM. The L-IX and L-X give rise to the long and short secondary paths. The long path projects as the ascending secondary tract to the ipsilateral nGS. The short path includes secondary fibers projecting to the motor layer of the L-X and the medullary reticular formation. Tertiary gustatory fibers arising in the nGS project ipsilaterally to two diencephalic nuclei: the nucleus glomerulosus and the nucleus diffusus lobi inferioris.

Studies in various vertebrate classes, particularly amphibians and mammals, have revealed that retinal ganglion cells with different functional properties project by means of axons of correspondingly different diameters onto specific target regions. Whether a similar pattern exists in teleosts is partly investigated in the present study. HRP was injected into the optic nerve of Sebastiscus and Navodon. The calibers of intraretinal HRP-labeled axons were classed as fine (ca. 0.8 micron), medium (ca. 1.3 micron), and coarse (ca. 2.5 microns). The calibers of HRP-labeled retinofugal axons were then determined in their target areas, and these can be summarized as follows: Optic hypothalamus: fine, medium. Lateral geniculate nucleus: fine. Dorsolateral thalamic nucleus: fine, medium. Area pretectalis: fine. Nucleus of the posterior commissure: fine, medium. Area ventralis lateralis, contralateral: fine, medium, coarse; ipsilateral: coarse. Optic tectum, stratum opticum: fine, medium; stratum fibrosum et griseum superficiale: fine, medium, coarse, segregated in sublayers; stratum album centrale: fine, medium, coarse. Therefore, fine fibers were found to reach all target areas except the ipsilateral area ventralis lateralis, and these were the only fibers found in the lateral geniculate nucleus, area pretectalis, and stratum griseum centrale of the optic tectum. Coarse fibers, on the other hand, were found only in the area ventralis lateralis and the optic tectum (stratum fibrosum et griseum superficiale and stratum album centrale). Terminal patterns of these fibers were also studied. Most fine fibers take tortuous courses giving off a few branches and terminate with many varicosities, and medium and coarse fibers give off several finer branches and terminate with bulbous swellings. The physiological significance of these findings is discussed. In addition, retrogradely labeled (retinopetal) cells were found in the olfactory bulb and the area ventralis pars ventralis of the telencephalon, as well as in the preoptic area and the dorsolateral thalamic nucleus.

Cerebellar efferent neurons in a teleost, Sebastiscus marmoratus, were studied by means of the horseradish peroxidase (HRP) tracing and Golgi impregnation methods. The cerebellar efferents or eurydendroid neurons are classified into two types (A and B) on the basis of the morphology of the dendrites, intracerebellar distribution, and other afferent connections. Type A neurons form a cell cluster in the lobus caudalis and have one thick primary dendrite from which several branches with sparse but long spines extend into the molecular layer of the lobus caudalis. Type B neurons, observed in both the valvula and corpus cerebelli, have two or three primary dendrites running along the ganglion cell layer, and the distal dendritic branches are distributed in molecular layer perpendicularly to the cerebellar surface. Small spines are densely studded on the dendrites of type B neurons. Somata of type A and B neurons are located in and just beneath the ganglion cell layer. The cerebellum projects to the ipsilateral nucleus lateralis valvulae and torus longitudinalis and bilaterally, but mostly contralaterally to the nucleus ventromedialis thalami of Schnitzlein ('62), nucleus ruber, the vicinity of the oculomotor complex, torus semicircularis, and brainstem reticular formation. Type A neurons send axons primarily to the vicinity of the oculomotor complex and partly to the nucleus ventromedialis thalami and the nucleus ruber, whereas type B cells project to all main cerebellar targets. In addition, type B cells are organized topographically in the cerebellum in relation to the efferent targets in the brainstem.

The neuronal connections in the central gustatory system of the crucian carp were examined by means of degeneration and HRP methods. Cell morphology in the primary gustatory lobes was studied in Golgi-impregnated material. Medium-sized neurons of the facial lobe emit axons which project to the secondary gustatory nucleus. The nucleus intermedius facialis of Herrick ('05) projects bilaterally. Large neurons send axons through the spinal trigeminal tract to terminate in the spinal trigeminal nucleus and in the medial funicular nucleus. In the vagal lobs, second-order neurons for the ascending projections are located in the superficial part of the sensory zone. These neurons project exclusively to the ipsilateral secondary gustatory nucleus. Neurons located in the deeper part of the sensory zone send axons to the motor zone and to the brainstem reticular formation to form short reflex arcs. The glossopharyngeal lobe has similar neuronal connections to the vagal sensory zone. Both facial and vagal lobes receive afferent projections from the following central structures: nucleus posterioris thalami, nucleus diffusus lobi inferioris, optic tectum, motor nucleus of the trigeminal nerve, medullary reticular formation, and the gray matter of the upper spinal cord. The facial lobe has an additional afferent from the mesencephalic reticular formation. The major sources to the medullary gustatory lobes are the nucleus posterioris thalami and nucleus diffusus lobi inferioris. Each type of neuron classified by morphology and location in the facial, glossopharyngeal, and vagal Jobes was correlated with its particular destination. Topographic projections were demonstrated in the secondary and tertiary gustatory centers.

The area postrema in goldfish is a dorsal midline structure in the caudal medulla spanning the level of the obex. As in other vertebrates, the sinus capillaries of the area postrema in goldfish are fenestrated. In goldfish, however, the area postrema is organized in a unique laminar fashion; from superficial to deep: meninx, vasculature, palisade layer, cell body layer, and ventral neuropil layer. Virtually all of the neurons of the area postrema exhibit tyrosine hydroxylase-like immunoreactivity. Each immunoreactive neuron is essentially bipolar, with a short apical dendrite extending dorsally to reach the external basal lamina of the capillaries and a basal dendrite reaching into the subjacent layer of neuropil. The apical dendrites have no synaptic specializations and probably function as interoceptors detecting blood-borne chemicals that leak out of the fenestrated capillaries. The basal dendrites receive synaptic input both within the neuropil of the area postrema and in the commissural nucleus of Cajal into which they extend. Primary afferent fibers of the subdiaphragmatic branches of the vagus nerve terminate within the area postrema and commissural nucleus. Thus the neurons of the area postrema may serve not only as direct chemoreceptive interoceptors but may also receive input from other visceral afferent systems.

The large majority of intraoral taste buds in goldfish are located on the gill arches and on the palatal organ, a muscular organ situated on the roof of the mouth. These taste buds are innervated by branches of the vagus nerve which terminate in a laminated vagal lobe, itself being an enlargement of the special visceral sensory column of the medulla. The tracer horseradish peroxidase (HRP) was used to determine the connectivity of the various branches of the vagus nerve that innervate the oropharyngeal gustatory surfaces. The entire oral cavity is mapped onto the vagal lobe so that the anterior end of the palatal organ and the most anterior gill arch are represented anteriorly in the vagal lobe; progressively more posterior oral structures are represented progressively more posteriorly in the lobe. The medial part of the palatal organ and the opposing gill arch surface, i.e., the ventromedial portion, are represented ventrally in the vagal lobe. The dorsolateral portions of the palatal organ and gill arches are represented dorsomedially in the vagal lobe. The topographic representation of the oral structures is similar for both the motor and sensory systems. In addition to this overall topographic organization, the different oropharyngeal structures are represented differentially in the layers of the vagal lobe. Palatal organ inputs reach layers VI and IX while gill arch inputs terminate in layers II, IV, and IX. The overall organization of the vagal lobe suggests a highly organized reflex system which is involved in the separation of food from substrate, especially during bottom feeding.

The coelomic root of the vagus nerve in goldfish is connected with sensory and motor nuclei of the medulla that are distinct from those serving the orobranchial roots of the same nerve. The primary sensory nucleus for coelomic sensation is itself divisible into medial and lateral subnuclei on the basis of afferent input and immunocytochemistry. The lateral subnucleus receives sensory input from the specialized chewing organ in the posterior pharynx and is poor in both substance P-like and tyrosine-hydroxylase-like immunoreactivities. The medial subnucleus receives input from the subdiaphragmatic gastrointestinal tract and is rich in substance P-like and tyrosine-hydroxylase-like immunoreactivities. The primary sensory fibers that innervate the gastrointestinal tract also project directly to the area postrema and to the vicinity of subdiaphragmatic visceral motor neurons. The vagal motor neuronal pool is divisible into three columns: paramedian (cardiac), medial, and lateral. The paramedian group innervates the heart and is situated in a loosely aggregated column at the boundary zone between the ventricular ependyma and the underlying brainstem. The medial vagal motor neurons innervate the subdiaphragmatic viscera, while the lateral column motor neurons innervate the posterior pharynx and muscles of the chewing organ. The motor neurons in this motor column are arranged in a topographic rostrocaudal order within the motor column according to the muscle of innervation. Thus both the general visceral sensory and general visceral motor nuclei of the medulla are organized into functional domains. Furthermore, in the goldfish, the special visceral (gustatory) and general visceral sensory nuclei form a continuous series in the medulla with the external and oral systems represented anteriorly and the pharyngeal and digestive systems represented posteriorly.

The primary gustatory sensory nuclei in catfish are grossly divisible into a vagal lobe and a facial lobe. In this study, the reflex connections of each gustatory lobe were determined with horseradish peroxidase (HRP) tracing methods. In addition, in order to determine the loci and morphology of the other brainstem cranial nerve nuclei, HRP was applied to the trigeminal, facial, glossopharyngeal, or vagus nerve. The sensory fibers of the facial nerve terminate in the facial lobe. The facial lobe projects bilaterally to the posterior thalamic nucleus, superior secondary gustatory nucleus, and medial reticular formation of the rostral medulla. The facial lobe has reciprocal connections with the lobobulbaris, medial reticular formation of the rostral medulla, descending trigeminal nucleus, medial and lateral funicular nuclei, and the vagal lobe, ipsilaterally; and with the facial lobe contralaterally. In addition, the facial lobe receives inputs from the raphe nuclei, from a pretectal nucleus, and from perilemniscal neurons located immediately adjacent to the ascending gustatory lemniscal tract at the level of the trigeminal motor nucleus. The gustatory fibers of the vagus nerve terminate in the vagal lobe, while the general visceral sensory fibers terminate in a distinct general visceral nucleus. The vagal lobe projects ipsilaterally to the superior secondary gustatory nucleus, lateral reticular formation, and n. ambiguus; and bilaterally to the commissural nucleus of Cajal. The vagal lobe has reciprocal connections with the ipsilateral lobobulbar nucleus and facial lobe, In addition, the vagal lobe receives input from neurons of the medullary reticular formation and perilemniscal neurons of the pontine tegmentum. In summary, the facial gustatory system has connections consonant with its role as an exteroceptive system which works in correlation with trigeminal and spinal afferent systems. In contrast, the vagal gustatory system has connections (e.g., with the n. ambiguus) more appropriate to a system involved in control of swallowing. These differences in central connectivity mirror the reports on behavioral dissociation of the facial and vagal gustatory systems.

Extrinsic and intrinsic fiber connections of the telencephalic subdivisions of Nieuwenhuys ('62) in a teleost, Sebastiscus marmoratus, were studied by means of horseradish peroxidase (HRP) and Fink-Heimer methods. The olfactory bulb projects bilaterally to area dorsalis pars posterior, area ventralis pars ventralis, pars lateralis, pars posterior, pars intermedia, and the nucleus posterior tuberis of Peter et al. ('75) and receives fibers from ipsilateral area dorsalis pars centralis, pars posterior, area ventralis pars dorsalis, and pars supracommissuralis. Area dorsalis pars posterior sends numerous fibers to the ipsilateral ventral region of area dorsalis pars medialis, from which fibers of the medial forebrain bundle arise and terminate in the inferior lobe and nucleus posterior tuberis. Area dorsalis pars lateralis, pars dorsalis, and the dorsal region of pars medialis are the main targets of extratelencephalic ascending afferents. Area dorsalis pars lateralis receives fibers from the ipsilateral nucleus prethalamicus of Meader ('34), where tectal projections terminate massively. Area dorsalis pars dorsalis and the dorsal region of pars medialis receive afferents from the ipsilateral nucleus preglomerulosus of Schnitzlein ('62), nucleus posterior tuberis, area preoptica pars medialis of Crosby and Showers ('69), and nucleus entopeduncularis of Sheldon ('12). Raphe nuclei and locus ceruleus project bilaterally to area dorsalis pars centralis, pars dorsalis, pars lateralis, and the dorsal region of pars medialis. Area dorsalis pars centralis, pars dorsalis, and the dorsal region of pars medialis are important sources of extratelencephalic efferents. These subdivisions give rise to the lateral forebrain bundle and project to the ipsilateral nucleus prethalamicus, nucleus preglomerulosus, inferior lobe, nucleus paracommissuralis of Ito et al. ('82), optic tectum, torus semicircularis, and the bilateral mesencephalic tegmentum. Within the telencephalon, most of the ventral subdivisions project to ipsilateral area dorsalis pars centralis, pars dorsalis, pars lateralis, and the dorsal region of pars medialis. Area dorsalis pars centralis has reciprocal connections with ipsilateral area dorsalis pars lateralis, pars dorsalis, pars posterior, and the dorsal region of pars medialis. A dorsal part of the anterior commissure is composed of axons of the ventral region of area dorsalis pars medialis destined to the contralateral ventral region of area dorsalis pars medialis. A ventral part of the anterior commissure contains axons of area dorsalis pars centralis destined to contralateral area dorsalis pars lateralis.