Extrinsic cells, immunoreactive to Ca-binding protein, as sources of thalamic visual centres in tortoises

Extrinsic sources of calcium-binding proteins involved in immunoreactive innervation of the visual thalamic nuclei Rot and GLd in turtles (Testudo horsfieldi and Emys orbicularis) were studied using HRP tracing method and immunohistochemistry. In 1.5-4.5 months after monocular enucleation calbindin (Calb)-, parvalbumin (Parv)- and calretinin (Calr)-labeling was found in fragments of degenerated retinal fibers in the contralateral optic tract and in some retinorecipient structures (optic tectum, GLd and GLv). Changes in GLd were detected in its neuropil part. in 2.0-3.5 months after unilateral ablation of tectum and pretectum, the densities of Parv-, Calb- and Aclr-immunoreactivity terminals and fibers were diminisched in the ipsilateral n. Rot, with the maximum effect seen in Parv. Following HRP injection into the visual thalamus (Rot and GLd), retrogradely labeled cells with Parv label only, were revealed in the ventrothalamic nucleus Enta, pretectal nucleus Ptv, and in all types of Ca-binding proteins (CaBPr) in separately labeled cells of the optic tectum. Thus, it has been shown that thalamic visual centers in turtles have multiple extrinsic cells, which serve as sources of CaBPr projections. The present data suggest that organization of CaBPr inputs to visual thalamus in reptiles (turtle) and higher amniotes are fundamentally similar.     

Calcium-binding proteins and cytochrome oxidase activity in the turtle optic tectum with special reference to the tectofugal visual pathway

Using immunohistochemistry and a tracer technique we investigated the distribution in the optic tectum of turtles (Emys orbicularis and Testudo horsfieldi) of the calcium-binding proteins (CaBPr) parvalbumin (PV), calbindin (CB) and calretinin (CR) before and after labeling of the nucleus rotundus (Rot) with horseradish peroxidase. The optic tectum activity of the cytochrome oxidase (CO) was studied in parallel. In the principal link of the tectofugal visual pathway (central gray layer, SGC) in both chelonian species, the sparse PV-ir as well as CB- and CR-ir neurons were found significantly varying both in number and the intensity of immunoreactivity of their bodies and dendrites. In contrast, the superficial (SGFS) and deeper periventricular (SGP) tectal layers comprised numerous cells immunoreactive to all three CaBPr in different proportions. Only few retrogradely labeled tectorotundal SGC neurons expressed PV, CB or CR. The very large PV-ir neurons in SGC and SAC were not retrogradely labeled; morphologically they matched the efferent neurons with descending projections. SGC neurons of two chelonian species differed in the level of CO activity. Intense immunoreactivity to all three CaBPr and high CO activity were detected in both species in SGFS neuropil with some differences in sublaminar distribution patterns. The peculiarities of the CaBPr and CO activity distribution patterns in different segments of SGC neurons are discussed as related to the laminar organization of the turtle tectum and its retinal innervation. It is suggested that in the projection tectorotundal SGC neurons the CaBPr are concentrated mainly in their distal dendrites that contact retinal afferents in the superficial retinorecipient tectal layer.     

Projections of Olfactory Bulbs and Non-Olfactory Telencephalic Structures in Amygdaloid Complex of the Turtle <Emphasis Type="Italic">Testudo horsfieldi</Emphasis>: A Study Using Anterograde Tracer Technique

To confirm the discrete character of projections of telencephalic olfactory and non-olfactory structures to the amygdaloid complex (AC) in the terrestrial turtle Testudo horsfieldi, a study was performed by the method of anterograde axonal transport of tracers (HRP, BDA). After a massive injection of the tracers into the main and accessory olfactory bulb, a dense accumulation of labeled fibers and terminals was found in ventral part of AC in the neuropil zones of nbam (J) and ncoam and very scanty—in nmam and ncam. After a massive injection of the tracers into non-olfactory telencephalic structures including dorsal cortex, pallidal enlargement, and ADVR, a very dense terminal field was observed in the dorsal AC part and a less dense one, with predominance of labeled fibers, in its ventral part. Local administration of the tracers separately into the dorsolateral (visual area) and the ventromedial (auditory-somatic area) parts of the ADVR allowed revealing discrete projections, respectively, to the laterocentral and mediocentral areas of the dorsal AC part with a relative overlapping in the central AC area. In all experiments, retrogradely labeled neurons in AC were also observed in zones of the corresponding bulbar and rostrotelencephalic projections. Thus, it has been shown that in the turtle AC there exist not only separation of direct olfactory and non-olfactory projections, but also discrete projections of different sensory areas of ADVR. Reciprocity of these connections is also confirmed. Organization of afferent olfactory and non-olfactory telencephalic connections in AC is similar in reptiles and in mammals.     

Central projections of the pineal complex in the silver lamprey Ichthyomyzon unicuspis

Summary The central projections of the pineal complex of the silver lamprey Ichthyomyzon unicuspis were studied by injection of horseradish peroxidase. The pineal tract courses caudally along the left side of the habenular commissure, and a few fibers penetrate the brain through the caudalmost portion of this commissure. Most of the fibers, however, continue caudally and enter the brain through the posterior commissure. The pineal tract projects bilaterally to the subcomissural organ, the superficial and periventricular pretectum, the posterior tubercular nucleus, the dorsal and ventral thalamus, the dorsal hypothalamus, the optic tectum, the torus semicircularis, the midbrain tegmentum, and the oculomotor nucleus. A few fibers decussate in the tubercular commissure, but the course of these decussate fibers could not be followed owing to the bilateral nature of the projections. No retrogradely labeled cells were found in the brain. With the exception of the projections to the optic tectum and torus semicircularis, the pineal projections in the silver lamprey are similar to those reported in other anamniote vertebrates.     

Retinal projections in the caecilian Ichthyophis kohtaoensis (Amphibia,Gymnophiona)

Summary The retinal projections of the caecilian Ichthyophis kohtaoensis were investigated by anterograde transport of HRP. The optic tract forms two bundles in the diencephalon, a narrow medial bundle in the optic tectum, and a basal optic tract consisting of few fibres. Terminal fields are in the thalamus, pretectum, tectum, and as a circum-scribed basal optic neuropile in the tegmentum. Thalamic, pretectal and tectal projections are contralateral as well as ipsilateral. The reduced but existing visual projection corresponds to a reduced but existing visually guided behaviour.     

Retinofugal projections in salamanders of the family Plethodontidae

Summary A comparison of the retinofugal projections in 14 species of plethodontid salamanders by means of the horseradish peroxidase (HRP) technique revealed almost identical contralateral projections. In all species studied three optic tracts were found. Behind the chiasma opticum the basal optic tract runs to the peduncle region, there forming the basal optic neuropil. The marginal optic tract courses from the chiasma over the thalamus to the tectum opticum where it covers the entire surface. In the anterior thalamus the marginal optic tract innervates the neuropil Bellonci-pars lateralis and the corpus geniculatum thalamicum, and more caudally the neuropil posterior thalami. The medial optic tract supplies the neuropil Bellonci-pars lateralis and pars medialis in the anterior thalamus from where it runs medial to the marginal optic tract as a separate tract to the uncinate field in the posterior thalamus.The ipsilateral projections show differences among the species studied, although the global organization remains constant. The differences mainly concern the marginal optic tract which varies from being weakly labeled and restricted to the rostral part of the tectum opticum, to being heavily labeled and innervating the entire tectum to its caudal edge. Species with the heaviest ipsilateral projections all belong to the plethodontid tribe Bolitoglossini, all of which show direct development, a highly projectile tongue, rather frontally oriented eyes and excellent depth perception. In these species the thalamic ipsilateral projection areas are equal in size and shape to the contralateral one. The ipsilateral projections to the tectum show two distinct layers, a superficial and a deep one, which intermingle with the contralateral projections. The two other ipsilateral tracts do not differ significantly among the plethodontid species: the medial optic tract is always heavily and the basal optic tract always weakly labeled.     

Enkephalin-immunoreactive cells in the mesencephalic tegmentum project to the optic tectum of the teleosts Salmo gairdneri and Salmo salar

Summary Immunocytochemistry using antibodies against Met-enkephalin and Leu-enkephalin has demonstrated a group of large enkephalin-immunoreactive neurons in the nucleus of the rostral mesencephalic tegmentum (mRMT) of two teleost fish, Salmo gairdneri and Salmo salar. Injections of cobalt-lysine in the medial optic tectum retrogradely labeled the above group of tegmental neurons. Tegmental neurons were labeled only ipsilaterally to the injection site. This indicates that enkephalinergic neurons in the nRMT project to the optic tectum, and that at least some of the enkephalinergic axons observed in the optic tectum belong to a tegmento-tectal pathway. Comparable enkephalinergic pathways have been described in reptiles and birds, where pretectal-mesencephalic nuclei contribute to the enkephalin-containing fibers that project to the optic tectum.     

Retinofugal and retinopetal connections in the upside-down catfish (Synodontis nigriventris)

Summary The retinofugal and retinopetal connections in the upside-down catfish Synodontis nigriventris were studied by use of the horseradish-peroxidase (HRP) techniques, autoradiography, and degeneration-silver methods. An unusual retinal projection to the torus semicircularis as well as projections to the retina from three different sources in the brain are described. After intra-ocular injections of HRP, labeled cells were found in the optic tectum, the dorsomedial optic nucleus and one of the pretectal nuclei. These new findings support the basic hypothesis (i) that neuronal connections are more extensive in primitive brains, and (ii) that the evolutionary development of more complex brains involves the loss of some selected connections.     

Experimental study of visual projection specificity in microphthalm and monophthalm embryos and adults of Gallus domesticus L.]

Research was carried out into the visual projections of embryos and chickens of Gallus domesticus L. who had undergone early optic vesicle removal and into microphthalmy or monophthalmy with ipsilateral optic fibres resulting from such removal. The architectonics of primary visual centres (nuclei ectomamillaris, geniculatus lateralis, lateralis anterior superficialis synencephali, griseus tectalis and the tectum opticum superficiale) and of the isthmo-opticus nucleus were compared with the architectonics of the same centres in anophthalms. From this research it can be seen that:--1. Optic fibres coming from limited ocular formation in microphthalms can reach the ectomamillaris nucleus in most cases and sustain existence; they may reach the tectum opticum without playing a qualitatively discernable morphogenetic role and act upon the isthmo-opticus nucleus. For these microphthalms, the nuclei lateralis anterior, geniculatus lateralis, superficialis synencephali and griseus tectalis are comparable to those of anophthalms. 2. Ipsilateral optic fibers can develop and show the same specificity and morphogenetic function as the microphthalms' optic fibres. 3. After hatching, some anophthalms shows an isthmo-opticus nucleus with scores of neurons. In general, observations during this research have shown that the specificity of microphthalms' optic fibres and ipsilateral optic fibres remain strictly the same whatever the operation under consideration.     

Lateral septal projections onto tubero-infundibular neurons in the hypothalamus of the guinea pig

Efferent projections of the lateral septal nucleus (LS) to the preoptic area and the hypothalamus were identified in 20 female guinea pigs after iontophoretic injection of the anterograde axonal tracer Fluoro-Ruby. Tubero-infundibular (TI) neurons of the preoptic area and the hypothalamus were retrogradely labeled after intracardiac injection of Granular Blue or Fluoro-Gold. Magnocellular neurons of the supraoptic and paraventricular nuclei were also labeled. The double labeling procedure allowed an estimation of the extent of the direct relationship between LS efferents and TI neurons. Contacts between lateral septal fibers and TI cell bodies were mainly observed at the light-microscopical level in the preoptic area. A group of labeled fibers coursing along the third ventricle established sparse connections with hypothalamic periventricular TI neurons. A few appositions was observed in the infundibular (arcuate) nucleus, suggestive of a monosynaptic regulation of TI neurons by a septo-arcuate tract. Close association with labeled magnocellular neurons was also noted at the edge of the supraoptic and paraventricular nuclei. The sparse but direct connections between LS and TI neurons may be involved in the neuroendocrine functions of the LS.     

Nervous connections of the parietal eye in adult Lacerta s. sicula Rafinesque as demonstrated by anterograde and retrograde transport of horseradish peroxidase

Summary Subsequent to the injection of horseradish peroxidase into the parietal eye of adult Lacerta sicula, the course of the parietal nerve and its projections were determined.The parietal nerve enters the left habenular ganglion where it branches into a medial and a lateral route. Some nerve fibers decussate within the habenular commissure. Whereas this pathway exhibits a striking asymmetry at the level of the habenular ganglia, its projections to the dorsolateral nucleus of the thalamus, the periventricular hypothalamic area, the preoptic hypothalamic and telencephalic regions, and the pretectal area are arranged in a strictly symmetric manner. A possible innervation of tegmental areas could not be proven due to the presence of endogenous peroxidase within these regions. No parietal nerve fibers were observed in the optic tectum.In a few animals investigated, scattered labeled perikarya were located in the periventricular hypothalamic gray indicating a parietopetal innervation in Lacerta sicula. The injection of horseradish peroxidase into one of the lateral eyes revealed terminal areas of the optic nerve within the preoptic region, and the thalamic and pretectal nuclei, displaying partial overlapping with the projections of the parietal nerve to these areas.From the present investigation further evidence is obtained that the pineal complex of lower vertebrates is a component of the photoneuroendocrine system. Particular emphasis is placed upon the nervous connections between the parietal eye and the hypothalamus, described for the first time in the present study.Supported by the Deutsche Forschungsgemeinschaft (Grant Ko 758/1)In partial fulfillment of the requirements of the degree of Dr. med., Faculty of Medicine, Justus Liebig University of Giessen     

Projections to the midbrain tectum in Salamandra salamandra L.

Following unilateral iontophoretic application of HRP into the optic tectum of Salamandra salamandra, retrogradely HRP-filled cells were found bilaterally in the pretectum, tegmentum isthmi, the reticular formation, pars medialis, and in the nucleus vestibularis magnocellularis. The area octavo-lateralis projects only to the caudal part of the tectum. Ipsilateral projections were noted from the dorsal gray columns of the cervical spinal cord, the dorsal tegmentum, the thalamus dorsalis pars medialis, thalamus dorsalis, pars anterior (to the rostral one-third of the tectum), the thalamus ventralis (in its entire rostro-caudal extent), and the preoptico-hypothalamic complex. Retrogradely filled cells were identified in deeper layers of the contralateral tectum. There are two telencephalic nuclei projecting ipsilaterally to the tectum via the lateral forebrain: the ventral part of the lateral pallium, and the posterior strioamygdalar complex.     

Bilateral retinal projections in the black piranah (Serrasalmus niger)

Summary The retinal projections were studied in the black piranah (Serrasalmus niger) with degeneration and autoradiographic methods. The projections are bilateral to the hypothalamic optic nucleus, the dorsomedial optic nucleus, corpus geniculatum ipsum of Meader (1934) and the optic tectum. Unilateral, crossed projections were traced to the pretectal nucleus and the cortical nucleus. The visual system of the black piranah is exceptionally well developed but has retained many primitive features including the extensive bilateral projections.     

Neurochemical organization of reptilian thalamus. Comparative analysis of amniote optical centers

Study of neurochemical characteristics of the turtle thalamus was performed using antibodies against several biologically active compounds; monoamines (5-HT, TH), neuropeptides (SP, m-Enk, NPY), against ChAT and by histochemical detection of NADPH-d. Based on our own results and literature data obtained on other representatives of reptiles, birds, and mammals, a comparative analysis was carried out of neurochemical organization of two thalamic optical centers—relay nuclei of thalamofugal (GLd) and tectofugal (Rot/LP-Pulv) systems of amniotes. Features of similarities and differences of these centers in representatives of non-mammalian and mammalian amniotes are revealed. GLd has a great similarity of the studied neurochemical characteristics in all amniotes. It receives innervation from all studied transmitter-modulatory systems with predominance of serotonin-, choline- and NPY-ergic projections. Neurochemical organization of Rot comparable with the tectorecipient part of mammalian LP-Pulv has a great resemblance in reptiles and birds, with considerable interspecies differences inside each class. In the majority of the studied systems, Rot is characterized by scant innervation. On the contrary, LP-Pulv receives sufficiently massive innervation from these systems. The most characteristic of this complex are rich SP- and cholinergic projections that are scant in Rot of reptiles and birds. The similar feature of Rot and LP-Pulv is the presence of massive serotonin- and NO-ergic (NADPH-positive) projections. The revealed similarities and differences of the neurochemical characteristics of thalamic optical centers among amniotes seem to reflect various transformations of thalamo- and tectofugal visual systems in the course of phylogenetic and adaptive evolution.     

Retinotectal Connexions of a Heterotopic Eye

THE mechanisms which cause the formation of specific synaptic connections in the nervous system are rather obscure. The development of specific connexions between eye and brain indicates a stage-dependent functional specification of the retina¹ which allows the retina to form predictable, specific connexions with the optic tectum, but the manner in which optic nerve fibres terminate in the tectum and the mechanisms which restore the visual projection after optic nerve regeneration are as yet not fully determined².     

Afferent connections of the optic tectum in channel catfish Ictalurus punctatus

Summary Horseradish peroxidase was injected unilaterally into the optic tectum of the channel catfish, Ictalurus punctatus. The sources of tectal afferents were thereby revealed by retrogradely labeled neurons in various brain centers. Retrogradely labeled cells were seen in both the ipsilateral and contralateral telencephalon. The superficial pretectal area was labeled on both sides of the brain. Ipsilateral projections were also observed coming from the entopeduncular nucleus. Both the anterior thalamic nucleus and the ventro-medial thalamic nucleus projected to the ipsilateral optic tectum. Cells in the ipsilateral nucleus of the posterior commissure were seen to project to the tectum. Labeled fibers were visualized in the lateral geniculate nucleus ipsilateral to the injected tectum, however, no labeled cell bodies were observed. Therefore, tectal cells project to the lateral geniculate nucleus, but this projection is not reciprocal. No labeled cells were found in the cerebellum. Labeled cells occurred in both the ipsilateral and contralateral medial reticular formation; they were also observed in the ipsilateral nucleus isthmi. A projection was seen coming from the dorsal funicular nucleus. Furthermore, labeled cells were shown in the inferior raphe nucleus.Abbreviations AP Area pretectalis - C Cerebellum - DPTN Dorsal posterior tegmental nucleus - H Habenula - IRF Inferior reticular formation - LI Inferior lobe - LGN Lateral geniculate nucleus - LR Lateral recess - MB Mammillary body - MRF Medial reticular formation - MZ Medial zone of the telencephalon - NC Nucleus corticalis - NDL-M Nucleus opticus dorsolateralis/pars medialis - NI Nucleus isthmi - NPC Nucleus of the posterior commissure - OPT Optic tectum - OT Optic tract - PC Posterior commissure - PN Pineal organ - PrOP Preoptic nucleus - PT Pretectum - TBt Tectobulbar tract - TEL Telencephalon - TL Torus longitudinalis - TS Torus semicircularis - VC Valvula cerebelli - VLTN Ventrolateral thalamic nucleus - VMTN Ventromedial thalamic nucleus     

Contralateral projections of the optic tectum in the zebra finch (Taenopygia guttata castanotis)

Summary Efferent projections of the optic tectum of zebra finches were investigated by injection of the radioactive anterograde tracer ³H-proline. In addition to a variety of ipsilateral projections, some contralateral connections were found. Quantitative evaluation of the recrossing tecto-rotundal and nucleus subpraetectalis/nucleus interstitio-praetecto-subpraetectalis projection revealed that these connections are much stronger than previously believed. In contrast, the tecto-tectal projection is very weak, as has been shown previously. Further support for this comes from results obtained using injections of retrograde tracers. The role of the different projections in conveying information from the ipsilateral eye to the ectostriatum, the telencephalic end-station of the tectofugal pathway, is discussed.     

Structure of visual pathways in the nervous system of freshwater pulmonate molluscs

Central nervous system of freshwater pulmonate molluscs Lymnaea stagnalis and Planorbarius corneus was stained using retrograde transport of neurobiotin in the optic tract fibers. In both species, perikarya and fibers of the stained neurons are found in all ganglia except the buccal ones. Afferent fibers of the optic nerve form dense sensory neuropil located in relatively small volume of cerebral ganglia. Typical neuronal groups sending their processes into the optic nerves of ipsilateral and contralateral body halves are described. Among them, neurons of visceral and parietal ganglia innervating both eyes concurrently as well as sending projections into peripheral nerves are revealed. These neurons, supposedly, have a function to integrate sensory signals, which may be a basis for regulation of light sensitivity of retina and functioning of peripheral organs. Bilateral links of the molluscan eye with the pedal ganglia cells and statocysts are found, which is, likely, a structural basis of certain known behavioral patterns related to stimulation of visual inputs in the studied gastropod molluscs.