Identification and characterization of visual pigments in caecilians (Amphibia: Gymnophiona), an order of limbless vertebrates with rudimentary eyes.

In comparison with the other amphibian orders, the Anura (frogs) and Urodela (salamanders), knowledge of the visual system of the snake-like Gymnophiona (caecilians) is relatively sparse. Most caecilians are fossorial with, as far as is known any surface activity occurring mainly at night. They have relatively small, poorly developed eyes and might be expected to possess detectable changes in the spectral sensitivity of their visual pigments. Microspectrophotometry was used to determine the spectral sensitivities of the photoreceptors in three species of caecilian, Rhinatrema bivittatum, Geotrypetes seraphini and Typhlonectes natans. Only rod opsin visual pigment, which may be associated with scotopic (dim light) vision when accompanied by other 'rod-specific' components of the phototransduction cascade, was found to be present. Opsin sequences were obtained from the eyes of two species of caecilian, Ichthyophis cf. kohtaoensis and T. natans. These rod opsins were regenerated in vitro with 11-cis retinal to give pigments with spectral sensitivity peaks close to 500 nm. No evidence for cone photoreception, associated with diurnal and colour vision, was detected using molecular and physiological methods. Additionally, visual pigments are short-wavelength shifted in terms of the maximum absorption of light when compared with other amphibian lineages.

The pathway controlling the pupillary light reflex in urodeles.

Although the photomechanical response of the iris in amphibians has been characterized, the central pathway of reflex control is unknown. We investigated this pathway by electrophysiological and anatomical techniques in urodeles. Sustained responding neurons in the pretectum were localized and characterized. Three neuron types were found and compared with tonic neurons in mammals. They were driven by the contralateral retina and their dendrites arborized in the pretectal neuropil. The localization of cell bodies resulted in the identification of a physiological nucleus comparable with the nucleus praetectalis olivaris. It was shown that the tonic neurons project ipsi- and contralaterally to the oculomotor region, where axons terminate in deep layers of the stratum album. Using horseradish peroxidase transport the ciliary ganglion in urodeles was identified. It occurs in ontogenesis parallel to the first detectable pupillary light reflex corresponding to the onset of metamorphosis.

Putative neurotransmitters in the retinae of three urodele species (Triturus alpestris, Salamandra salamandra, Pleurodeles waltli).

The immunocytochemical localization of several substances with putative neurotransmitter or modulator properties was investigated in the retinae of three urodele species. Gamma-aminobutyric acid-like immunoreactive labelling appeared in different types of amacrine and horizontal cells. In addition, labelled fibres in the optic nerve were detected. It was not possible to determine whether these fibres were ganglion-cell axons or part of an efferent projection. Endogenous serotonin was found in several populations of amacrine cells including stratified and diffuse types. Glucagon-like immunoreactivity appeared in one bistratified amacrine cell type, and neurotensin-like immunoreactivity was detected in a single monostratified amacrine cell type. Metenkephalin-like-immunoreactive labelling was type. Metenkephalin-like-immunoreactive labelling was rare but found in several sublaminae of the inner plexiform layer. Thus each peptide-like-immunoreactive cell type makes up a distinct and unique population of cells and probably has a special functional role in retinal processing. There are striking similarities in the peptide-like immunoreactive patterns of Triturus alpestris and Necturus maculosus whereas in Ambystomatidae the peptide-like-immunoreactive systems appear to be differently organized. This supports the hypothesis that Salamandridae and Proteidae are more closely related to each other than to the Ambystomatidae.

On the presence of nucleus ruber in the urodele Salamandra salamandra and the caecilian Ichthyophis kohtaoensis.

The presence of nucleus ruber in urodeles and caecilians (amphibia) was investigated. For that purpose, horseradish peroxidase was applied to the rostral spinal cord, the medulla oblongata at various levels and the dorsolateral funiculus. Whereas Salamandra salamandra possesses a rubrospinal tract, it is absent in the limbless caecilian Ichthyophis kohtaoensis.

Two populations of serotonin-immunoreactive neurons in the frog (Rana esculenta) retina.

Serotonin immunocytochemical staining in retina whole-mounts of Rana esculenta revealed two differently stained amacrine cell populations. Statistical investigations using nearest neighbor analysis and comparison with different theoretical cell distributions suggested that the two serotonin-immunoreactive cell populations are related to functionally different types. The analysis of type A (intensely stained somata, less numerous) revealed a good coincidence with the normal Gaussian distribution indicating a functionally homogeneous population. The distribution of the neurons of type B (weakly stained somata, more numerous) reflects rather a random dot pattern than a regular arrangement. Thus there is evidence that this last type is a heterogeneous cell population consisting of several functionally different types.

Topologic and connectional analysis of the dorsal thalamus of Triturus alpestris (amphibia, urodela, salamandridae).

A morphological and connectional analysis was performed on the dorsal thalamus of the alpine newt, Triturus alpestris. We have used a graphic reconstruction technique for the evaluation of the connectional (HRP) data. On the basis of these reconstructions, we propose a subdivision of the salamandrid dorsal thalamus into subhabenular, anteroventral, and posterodorsal zones. Each of these zones is defined by its telencephalic projections ("ascending thalamofugal systems"). The posterodorsal zone projects to the striatum, the anteroventral zone to the pallium. The subhabenular zone projects to the subpallial telencephalon and to the tegmentum. This zonal subdivision allows a more detailed comparison of the salamandrid dorsal thalamic features with ranid dorsal thalamic structures. We compare our dorsal thalamic zones to the ones proposed by Herrick (J. Comp. Neurol. 62:239-261, '35, The Brain of the Tiger Salamander. Chicago: The University of Chicago Press, '48). Furthermore, using the same reconstructive technique, we undertook an analysis of the spatial relations of various inputs to the salamandrid dorsal thalamus ("thalamopetal systems"). Besides the well-known retinal inputs, we identified the tectum and the tegmentum as sources of inputs to the thalamus. We provide evidence that there is no extensive multi- or unimodal overlap of these thalamopetal systems.

Localization of motoneurons innervating the extraocular muscles in Salamandra salamandra L. (Amphibia, Urodela).

The central innervation patterns of the extraocular muscles were investigated in the European fire salamander Salamandra salamandra L. by means of the horseradish peroxidase method. The ipsilateral portion of the nucleus nervi oculomotorii, which is located in the rostral ventral tegmentum mesencephali, supplies the musculi recti inferior and medialis and the musculus obliquus inferior without a clear somatotopic representation of the motoneurons. The musculus rectus superior is innervated mainly by a contralateral portion of this nucleus. A definite nucleus Edinger-Westphal could not be recognized. The nucleus nervi trochlearis, which rostrally joins the nucleus nervi oculomotorii with a gap of only about 40 micron between the nuclei, is situated completely contralateral to the musculus obliquus superior supplied by it. The nucleus nervi abducentis, innervating the musculus rectus lateralis, and the nucleus accessorius nervi abducentis, supplying the musculus retractor bulbi, are found in the ipsilateral medulla oblongata and exhibit a large rostrocaudal extension from the eighth cranial nerve to the first root of the vagus nerve. Dendrites of the nucleus nervi oculumotorii and of the nucleus accessorius nervi abducentis extend into neuropil areas receiving primary sensory afferents.

Two thalamo-telencephalic pathways in a urodele, Triturus alpestris.

Ascending thalamo-telencephalic projection systems have been investigated in an urodele, Triturus alpestris, using the horseradish peroxidase technique. Two separate dorsal thalamic projections onto the telencephalon have been identified; one arises from the posterior dorsal thalamus and terminates in the ipsilateral striatum, the other originates from anterior dorsal thalamic cells and reaches the medial pallium and a part of the dorsal pallium bilaterally. Both systems, which are spatially well segregated, might carry visual information to the telencephalon, as the posterior dorsal thalamus receives tectal, and the anterior dorsal thalamus direct retinal input. The urodele projection scheme as described here shows great similarities to the one described in anurans, although there are remarkable cytoarchitecture differences between the anuran and the urodele thalamus.

Amphibian optokinetic after nystagmus: properties and comparative analysis in various species.

Optokinetic nystagmus and after nystagmus were studied in six amphibian species, three urodeles and three anurans. It was demonstrated that two of the urodeles, Hydromantes italicus and Salamandra salamandra, display a relatively well-developed optokinetic after nystagmus, which is less pronounced in Bombina variegata and nearly absent in Bufo bufo, Rana temporaria, and Tylototriton verrucosus. These results indicate a certain degree of velocity storage in the optokinetic reflex of some amphibians.

Visual projections in larval Ichthyophis kohtaoensis (Amphibia: gymnophiona).

The visual projection patterns of retinal efferents were studied in larval Ichthyophis kohtaoensis by means of anterogradely transported HRP. Our results show in all larvae a projection contralateral to a thalamic terminal field, a pretectal terminal field, and a basal optic neuropil, but only a sparse innervation of the contralateral tectum. In addition, all larvae possess an uncrossed projection to a thalamic and a pretectal terminal field. The fibers are bilaterally almost confined to the medial optic tract with only a few fibers running in the marginal and basal optic tract. The ipsilateral and contralateral tracts and terminal fields seem to enlarge during larval life. Comparison with other amphibian orders reveals that larval Ichthyophis are unique in that they develop the medial optic tract and the related thalamic and pretectal terminal fields very early in larval life. In addition they possess only a very sparse tectal projection, though it is the largest projection in larval urodeles and anurans. This suggests a selective phylogenetic loss of those ganglion cells or collaterals which project mainly to the tectum in other amphibian orders and a change in the ontogenetic program leading to an earlier development of the medial optic tract in Ichthyophis as compared to urodeles and anurans.

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

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 circumscribed 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.

Eye accommodation during prey capture behaviour in salamanders (Salamandra salamandra L.).

The eyes of fire salamanders were examined refractometrically before, during and after prey capture behaviour. It was ascertained that the refraction makes a myopic shift before the salamander turns to the prey. This means that fire salamanders use accommodation for distance estimation.

Pretectal neurons project to the salamander retina.

Injection of horseradish peroxidase solution into the eye of some european salamander species labeled retrogradely pretectal neurons. These neurons lie near the commissura posterior and extend their dendrites into the visual pretectal neuropil. On the average, each animal contained 28 labeled cells. Their distribution is bilaterally with a slight preference to the contralateral pretectum. As in other vertebrates these retrogradely labeled pretectal cells may be considered as efferents to the salamander retina.

Development of retinofugal neuropil areas in the brain of the alpine newt, Triturus alpestris.

The development of the retinofugal projection areas of the brain has been studied in larvae of Triturus alpestris by means of anterograde transported horseradish peroxidase. The optic tract establishes contacts with the optic tectum prior to the onset of robust terminal formation in the diencephalon. The tectum becomes covered by the retinofugal projection in a rostro-caudal direction. The basal optic neuropil develops synchronously with the oculomotor neurons. Their dendrites extend into this neuropil area. A small amount of uncrossed label occurs long before metamorphosis. Around metamorphotic climax this ipsilateral label increases but does not attain the adult pattern even three monts postmetamorphosis. The data are compared with the onset of visual induced behaviour.

Projections of color coding retinal neurons in urodele amphibians.

Optic fiber projection in the brain of Salamandra salamandra was investigated by degeneration techniques. Terminal fields are described in the thalamus and in the optic tectum. Microelectrode recordings were performed from ganglion cells in the retina and from their terminals in the thalamus and tectum in Salamandra and Triturus alpestris. 'On' cells showed maximal sensitivity either in the blue or in the yellow spectral region; they project to the thalamus. Color coding 'on-off' cells project to the tectum opticum. In Triturus a seasonal change in these neurons occurs. Probably due to transition of vitamin A2 into vitamin A1 the spectral sensitivity is different. In springtime blue-red opponent-color neurons were recorded, in fall however, blue-yellow neurons were found.

Color vision in salamander larvae.

In behavioral experiments using monochromatic prey-patterns, larval salamanders (Salamandra salamandra) are able to discriminate colored from white light. In the retina only blue-yellow opponent-color ganglion cells were recorded. Thus the color vision in these animals is dichromatic.