Julián Yáñez

Experimental study of the connections of the telencephalon in the rainbow trout (Oncorhynchus mykiss). II: Dorsal area and preoptic region

In this study and the accompanying article (Folgueira et al., 2004a ), the fluorescent carbocyanine dye 1,1′‐dioctadecyl 3,3,3′,3′‐tetramethylindocarbocyanine perchlorate (DiI) was used in fixed tissue to comprehensively analyze the connections of the different regions of the telencephalic lobes and the preoptic region of the rainbow trout. Here, we analyze the connections of the dorsal area (D; pallium) of the telencephalon, and the preoptic region, as well as the telencephalic connections of several structures in the diencephalon and brainstem of juvenile trout. The dorsal plus dorsolateral pallial zone of D (Dd+Dl‐d) receives afferents from contralateral Dd+Dl‐d, the ventral area of the telencephalon, preoptic nucleus, suprachiasmatic nucleus, medial thalamus, preglomerular complex, anterior and lateral tuberal nuclei, posterior tuberal nucleus, posterior hypothalamic lobe, superior raphe nucleus, and the rhombencephalic central gray and reticular formation, and projects to the central zone of D (Dc), medial thalamus, and some caudomedial hypothalamic regions. The medial zone of D (Dm) maintains reciprocal connections with the preglomerular complex and also receives afferents from the preoptic nucleus, suprachiasmatic nucleus, anterior tuberal nucleus, preglomerular tertiary gustatory nucleus, posterior tubercle, superior raphe nucleus, locus coeruleus, and the rhombencephalic central gray, and reticular formation. Dc receives fibers mainly from Dd+Dl‐d, preoptic nucleus, preglomerular complex, and torus semicircularis and projects to several extratelencephalic centers, including the paracommissural nucleus, optic tectum, torus semicircularis, thalamus, preglomerular complex, posterior tubercle nuclei, and inferior hypothalamic lobes. The posterior zone of D (Dp) is mainly connected with the olfactory bulbs, the ventral and supracommissural nuclei of the ventral area (subpallium), the preoptic nucleus, and the preglomerular complex and projects to wide hypothalamic and posterior tubercular regions. The preoptic nucleus projects to the olfactory bulb, to most regions of the telencephalic lobes, and to several diencephalic and brainstem structures. These results reveal complex and specialized connectional patterns in the rainbow trout dorsal telencephalon and preoptic region. Most of these connections have not been described previously in salmonids. These connections indicate that the salmonid telencephalon is involved in multisensorial processing and modulation of brain activity. J. Comp. Neurol. 480:204–233, 2004.

Distribution of choline acetyltransferase (ChAT) immunoreactivity in the brain of the adult trout and tract-tracing observations on the connections of the nuclei of the isthmus.

The distribution of cholinergic neurons and fibers was studied in the brain and rostral spinal cord of the brown trout and rainbow trout by using an antiserum against the enzyme choline acetyltransferase (ChAT). Cholinergic neurons were observed in the ventral telencephalon, preoptic region, habenula, thalamus, hypothalamus, magnocellular superficial pretectal nucleus, optic tectum, isthmus, cranial nerve motor nuclei, and spinal cord. In addition, new cholinergic groups were detected in the vascular organ of the lamina terminalis, the parvocellular and magnocellular parts of the preoptic nucleus, the anterior tuberal nucleus, and a mesencephalic tegmental nucleus. The presence of ChAT in the magnocellular neurosecretory system of trout suggests that acetylcholine is involved in control of hormone release by neurosecretory terminals. In order to characterize the several cholinergic nuclei observed in the isthmus of trout, their projections were studied by application of 1,1`‐dioctadecyl‐3,3,3`,3`‐tetramethylindocarbocyanine perchlorate (DiI) to selected structures of the brain. The secondary gustatory nucleus projected mainly to the lateral hypothalamic lobes, whereas the nucleus isthmi projected to the optic tectum and parvocellular superficial pretectal nucleus, as previously described in other teleost groups. In addition, other isthmic cholinergic nuclei of trout may be homologs of the mesopontine system of mammals. We conclude that the cholinergic systems of teleosts show many primitive features that have been preserved during evolution, together with characteristics exclusive to the group. J. Comp. Neurol. 428:450–474, 2000.

Afferent and efferent connections of the habenula in the rainbow trout (Oncorhynchus mykiss): An indocarbocyanine dye (DiI) study

The habenula is a conserved structure in the brain of vertebrates. With the aim of further understanding of the evolution of the habenular system in vertebrates, we studied the afferent and efferent connections of the habenula of the rainbow trout. Experiments included application of the carbocyanine dye 1,1′‐dioctadecyl‐3,3,3′,3′‐tetramethylindocarbocyanine perchlorate (DiI) into the habenula, telencephalon, pineal organ, posterior tubercle, and interpeduncular nucleus (IPN). The results obtained reveal a consistent pattern of habenular connections. Most afferents originate from three nuclei, one extending from the preoptic region to the rostral thalamus (the entopeduncular nucleus), the second located in the region of the hypothalamus‐posterior tubercle and consisting of large bipolar cells (tuberculohabenular nucleus), and the third in the preoptic region (preoptic nucleus). A few large neurons of the locus coeruleus appeared to be labeled in some cases. The trout habenula also receives pineal and parapineal projections. Small labeled glial cells were observed in the thalamus around the fasciculus retroflexus and, sometimes, around the IPN. The most conspicuous efferents coursed in the fasciculus retroflexus to the IPN, the isthmal raphe, and the central gray. The existence of olfactohabenular or habenulotelencephalic projections is discussed.

An experimental study of the connections of the telencephalon in the rainbow trout (Oncorhynchus mykiss). I: Olfactory bulb and ventral area

The fluorescent carbocyanine dye 1,1′‐dioctadecyl 3,3,3′,3′‐tetramethylindocarbocyanine perchlorate (DiI) was used in fixed tissue to comprehensively analyze the connections of the olfactory bulbs and the different regions of the ventral (V) area of the telencephalic lobes (subpallium) of the rainbow trout. With this goal, DiI was applied to the different telencephalic nuclei and zones, as well as to the olfactory nerve, the olfactory bulb, the retina, and to several structures in the diencephalon and brainstem of juvenile trout. The olfactory bulbs maintain reciprocal connections with several regions of the telencephalon [ventral nucleus of V (Vv), supracommissural nucleus (Vs), posterior zone of D (Dp), preoptic nucleus], and also project to the diencephalon (posterior tuberal nucleus, posterior hypothalamic lobe). Vv receives afferents from Vs, the dorsal nucleus of V (Vd), the preoptic nucleus, and from several nuclei in the diencephalon and brainstem (suprachiasmatic nucleus, anterior and lateral tuberal nuclei, preglomerular complex, tertiary gustatory nucleus, posterior tubercle, inferior hypothalamic lobes, thalamus, torus semicircularis, secondary gustatory nucleus, locus coeruleus, superior raphe nucleus, central gray, and reticular formation), and projects to dorsal (pallial) regions and most of the nuclei afferent to Vv. The dorsal nucleus of V (Vd) and Vs mainly project to the dorsal area. In an accompanying article (Folgueira et al., 2004 ), we present the results of application of DiI to dorsal (pallial) telencephalic regions, as well as of several experiments of tracer application to extratelencephalic regions. The results presented here, together with those of the accompanying article, reveal a complex connectional pattern of the rainbow trout ventral telencephalon, most of these connections having not been described previously in salmonids. J. Comp. Neurol. 480:180–203, 2004.

AFFERENT AND EFFERENT CONNECTIONS OF THE PARAPINEAL ORGAN IN LAMPREYS : A TRACT TRACING AND IMMUNOCYTOCHEMICAL STUDY

The neural connections of the parapineal organ of two species of lampreys were studied with the fluorescent dye 1,1′‐dioctadecyl‐3,3,3′,3′‐tetramethylindocarbocyanine perchlorate (DiI) and with immunocytochemistry. The lamprey parapineal organ consists of a vesicle and a ganglion that are connected to the left habenula. Labeling experiments included the application of DiI to the parapineal organ, left and right fasciculus retroflexus, left habenula, and the left pretectal region. Afferent parapineal fibers run in the left fasciculus retroflexus to the interpeduncular nucleus. The parapineal fibers of this fascicle arose from parapineal ganglion cells, whereas DiI application to the left habenula labeled both neurons of this ganglion and bipolar cells in the parapineal vesicle. Efferent neurons were observed in the left habenula, and bilaterally in the subhippocampal nucleus and the dorsal pretectum. Labeling with DiI also revealed a hippocampal projection.

Early development of the retina and pineal complex in the sea lamprey: Comparative immunocytochemical study

Lampreys have a complex life cycle, with largely differentiated larval and adult periods. Despite the considerable interest of lampreys for understanding vertebrate evolution, knowledge of the early development of their eye and pineal complex is very scarce. Here, the early immunocytochemical organization of the pineal complex and retina of the sea lamprey was studied by use of antibodies against proliferating cell nuclear antigen (PCNA), opsin, serotonin, and γ‐aminobutyric acid (GABA). Cell differentiation in the retina, pineal organ, and habenula begins in prolarvae, as shown by the appearance of PCNA‐negative cells, whereas differentiation of the parapineal vesicle was delayed until the larval period. In medium‐sized to large larvae, PCNA‐immunoreactive (‐ir) cells were numerous in regions of the lateral retina near the differentiated part of the larval retina (central retina). A late‐proliferating region was observed in the right habenula. Opsin immunoreactivity appears in the pineal vesicle of early prolarvae and 3 or 4 days later in the retina. In the parapineal organ, opsin immunoreactivity was observed only in large larvae. In the pineal organ, serotonin immunoreactivity was first observed in late prolarvae in photoreceptive (photoneuroendocrine) cells, whereas only a few of these cells appeared in the parapineal organ of large larvae. No serotonin immunoreactivity was observed in the larval retina. GABA immunoreactivity appeared earlier in the retina than in the pineal complex. No GABA‐ir perikaryon was observed in the retina of larval lampreys, although a few GABA‐ir centrifugal fibers innervate the inner retina in late prolarvae. First GABA‐ir ganglion cells occur in the pineal organ of 15–17 mm larvae, and their number increases during the larval period. The only GABA‐ir structures observed in the parapineal ganglion of larvae were afferent fibers, which appeared rather late in development. The time sequence of development in these photoreceptive structures is rather different from that observed in teleosts and other vertebrates. This suggests that the unusual development of the three photoreceptive organs in lampreys reflects specialization for their different functions during the larval and adult periods. J. Comp. Neurol. 442:250–265, 2002.

Experimental study of the connections of the gustatory system in the rainbow trout, Oncorhynchus mykiss.

Salmonids are a group of teleosts with a nonspecialized gustatory system. With the aim of describing the gustatory connections in a member of this group, we carried out tract‐tracing experiments using the lipophilic carbocyanine dye 1,1′‐dioctadecyl 3,3,3′,3′‐tetramethylindocarbocyanine perchlorate (DiI) in fixed brains of the rainbow trout (Oncorhynchus mykiss). The neural tracer was applied to the primary viscerosensory column, secondary gustatory visceral nucleus (SGN), torus lateralis (TL), and tertiary gustatory nucleus (TGN), the dorsal part of the ventral area of the telencephalon (dorsal‐Vv), and the medial area of the dorsal telencephalon (Dm). The primary viscerosensory column projects mainly to the SGN. DiI application to the SGN showed a bilateral and reciprocal connection with the TGN and a rostral portion of the nucleus of the lateral hypothalamic recess. The application of DiI in the dorsal‐Vv and Dm at levels rostral to the anterior commissure led to labeling of a restricted group of diencephalic neurons in the TGN and sending dendrites to the TL. DiI application to the TL/TGN anterogradely labeled fibers that coursed in the medial forebrain bundle innervating the precommissural portion of the dorsal‐Vv and Dm. Caudally, this type of application led to labeling of fibers in the viscerosensory column and perikarya in the SGN. Tract‐tracing results showed direct projections from the diencephalic and rhombencephalic gustatory nuclei to the telencephalon. There was a direct and reciprocal connection between the SGN and the ventral telencephalon. The results showed that the gustatory connections of the trout are similar to those of teleosts, with highly specialized gustatory centers as in cyprinids and ictalurids, and to that observed in the percomorph tilapia, thus demonstrating a basic organization that is shared by most teleosts. J. Comp. Neurol. 465:604–619, 2003.

Cholinergic and GABAergic neuronal elements in the pineal organ of lampreys, and tract-tracing observations of differential connections of pinealofugal neurons.

Abstract The putative cholinergic and GABAergic elements of the pineal organ of lampreys were investigated with immunocytochemistry to choline acetyltransferase (ChAT) and γ-aminobutyric acid (GABA), and by acetylcholinesterase (AChE) histochemistry. For comparison we also carried out immunocytochemistry to serotonin (5-HT) and a tract-tracing investigation of the two types of projecting cells, i.e., ganglion cells and long-axon photoreceptors. Most photoreceptors were ChAT-immunoreactive (ChAT-ir) and AChE-positive, while ganglion cells and the pineal tract were ChAT-negative and AChE-negative or only faintly positive. These results strongly suggest the presence of a cholinergic system of photoreceptors in the lamprey pineal organ. GABA-ir fibers that appear to originate from faintly to moderately stained ganglion cells were observed in the pineal stalk. Immunocytochemistry to 5-HT indicated the presence of two types of 5-HT-ir cells, bipolar cells and ganglion-like cells. The connections of the ganglion cells and long-axon photoreceptors were also studied by application of DiI to the pineal stalk in fixed brains or of biotinylated dextran amine (BDA) to one of the main targets of pinealofugal fibers (optic tectum or mesencephalic tegmentum) in isolated brains in vitro. Some long-axon photoreceptors and ganglion cells were labeled from the optic tectum. However, BDA application to the tegmentum exclusively labeled ganglion cells in the pineal organ. These results indicate that the two morphological types of afferent pineal neuron have different projections. No labeled cells were observed in the parapineal organ in BDA experiments, indicating that this organ and the pineal organ are involved in different neural circuits.

Afferent and efferent connections of the cerebellum of a salmonid, the rainbow trout (Oncorhynchus mykiss): A tract-tracing study

The connections of the cerebellum of the rainbow trout were studied by experimental methods. The pretectal paracommissural nucleus has reciprocal connections with the cerebellum. Three additional pretectal nuclei project to both the corpus and valvula cerebelli, and seem to receive cerebellar afferents. A large number of cells of the lateral nucleus of the valvula project to wide regions of the cerebellum, including the valvula, the corpus, the granular eminences, and the caudal lobe, whereas the contralateral inferior olive and scattered reticular cells project only to the corpus and valvula cerebelli. Afferents to the corpus were also observed from the ventral tegmental nucleus, the “paraisthmic nucleus,” the perilemniscal nucleus, the central gray, and the octavolateral area. Valvular afferents were also observed from the torus semicircularis and the midbrain tegmental areas. In most cases of cerebellar application, labeled fibers were seen in the thalamus, the pretectum, the torus longitudinalis and torus semicircularis, the nucleus of the medial longitudinal fascicle, and midbrain and rhombencephalic reticular areas. From the corpus cerebelli some fibers also project to the posterior tubercle and the hypothalamus. Moreover, the granular eminences project to the cerebellar crest. DiI application to most of the areas showing labeled fibers after cerebellar tracer application led to the labeling of characteristic eurydendroid cells, mainly in the valvula cerebelli and the caudal lobe. A few putative eurydendroid cells were labeled from the octavolateralis regions. These results in a teleost with a generalized brain indicate several differences with respect to the cerebellar connections reported in other teleost fishes that have specialized brains. J. Comp. Neurol. 497:542–565, 2006.

Calbindin and calretinin immunoreactivities in the retina of a chondrostean, Acipenser baeri

Abstract. The distribution of calbindin and calretinin in the retina of the sturgeon Acipenser baeri was studied with immunocytochemistry. Western blot analysis of brain extracts, together with immunocytochemical results in the retina and brain, indicated the presence of the two calcium-binding proteins in sturgeon. Calbindin immunocytochemistry revealed only a large displaced bipolar cell type with narrowly stratified axons, similar to some mixed rod and cones bipolar cells described in teleosts. The plexus formed by the axons of these cells in the inner plexiform sublayer was similar to that formed by calbindin-immunoreactive diffuse bipolar cells of some mammals. Calretinin immunocytochemistry also stained these displaced bipolar cells, most ganglion cells including displaced ganglion cells (Dogiel cells), and some amacrine cells of the inner nuclear layer. The distribution of calbindin and calretinin immunoreactivities in the retina of a primitive bony fish indicates that these proteins are highly specific to the cell type.