Core-and-belt organisation of the mesencephalic and forebrain auditory centres in turtles: expression of calcium-binding proteins and metabolic activity.

The distribution of immunoreactivity to the calcium-binding proteins parvalbumin, calbindin and calretinin and of cytochrome oxidase activity was studied in the mesencephalic (torus semicircularis), thalamic (nucleus reuniens) and telencephalic (ventromedial part of the anterior dorsal ventricular ridge) auditory centres of two chelonian species Emys orbicularis and Testudo horsfieldi. In the torus semicircularis, the central nucleus (core) showed intense parvalbumin immunoreactivity and high cytochrome oxidase activity, whereas the laminar nucleus (belt) showed low cytochrome oxidase activity and dense calbindin/calretinin immunoreactivity. Within the central nucleus, the central and peripheral areas could be distinguished by a higher density of parvalbumin immunoreactivity and cytochrome oxidase activity in the core than in the peripheral area. In the nucleus reuniens, the dorsal and ventromedial (core) regions showed high cytochrome oxidase activity and immunoreactivity to all three calcium-binding proteins, while its ventrolateral part (belt) was weakly immunoreactive and showed lower cytochrome oxidase activity. In the telencephalic auditory centre, on the other hand, no particular region differed in either immunoreactivity or cytochrome oxidase activity. Our findings provide additional arguments in favour of the hypothesis of a core-and-belt organisation of the auditory sensory centres in non-mammalian amniotes though this organisation is less evident in higher order centres. The data are discussed in terms of the evolution of the auditory system in amniotes.

Preoptic FMRF-amide-like immunoreactive projections to the retina in the lamprey (Lampetra fluviatilis).

A centrifugal visual system showing FMRF-amide-like immunoreactivity has been demonstrated in Lampetra fluviatilis by using immunocytochemical and hodological techniques. From 50 to 60 immunoreactive neurons, labelled after contralateral intraocular injection of rhodamine beta-isothiocyanate, form a small, clearly defined, nucleus in the lateral neural plate of the magnocellular preoptic nucleus. These cells give rise to immunoreactive axons which have been observed at the base of the nucleus, in the optic chiasma and in the optic nerve, to project into the intermediate plexiform layer of the retina, which separates the layer of internal horizontal cells from the layer of external horizontal cells. This FMRF-amide-like immunoreactive centrifugal visual system is compared to that described in Gnathostomes.

Synaptic circuitry in the retinorecipient layers of the optic tectum of the lamprey (Lampetra fluviatilis). A combined hodological, GABA and glutamate immunocytochemical study.

The ultrastructure of the retinorecipient layers of the lamprey optic tectum was analysed using tract tracing techniques combined with GABA and glutamate immunocytochemistry. Two types of neurons were identified; a population of large GABA-immunonegative cells, and a population of smaller, highly GABA-immunoreactive interneurons, some of whose dendrites contain synaptic vesicles (DCSV). Five types of axon terminals were identified and divided into two major categories. The first of these are GABA-immunonegative, highly glutamate-immunoreactive, contain round synaptic vesicles, make asymmetrical synaptic contacts, and can in turn be divided into AT1 and AT2 terminals. The AT1 terminals are those of the retinotectal projection. The origin of the nonretinal AT2 terminals could not be determined. AT1 and AT2 terminals establish synaptic contacts with DCSV, with dendrites of the retinopetal neurons (DRN), and with conventional dendritic (D) profiles. The terminals of the second category are GABA-immunoreactive and can similarly be divided into AT3 and AT4 terminals. The AT3 terminals contain pleiomorphic synaptic vesicles and make symmetrical synaptic contacts for the most part with glutamate-immunoreactive D profiles. The AT4 terminals contain rounded synaptic vesicles and make asymmetrical synaptic contacts with DRN, with DCSV, and with D profiles. A fifth, rarely observed category of terminals (AT5) contain both clear synaptic vesicles and a large number of dense-core vesicles. Synaptic triads involving AT1, AT2 or AT4 terminals are rare. Our findings are compared to these of previous studies of the fine structure and immunochemical properties of the retinorecipient layers of the optic tectum or superior colliculus of Gnathostomes.

Fine structure of the visual dorsolateral anterior thalamic nucleus of the pigeon (Columba livia): a hodological and GABA-immunocytochemical study.

The ultrastructure of the lateroventral subcomponent of the visual dorsolateral anterior thalamic nucleus of the pigeon (DLLv) was analyzed using hodological techniques and GABA-immunocytochemistry. Two types of GABA-immunonegative hyperpalliopetal neurons and a single type of strongly GABA-immunoreactive (-ir) interneuron were identified, the latter displaying long dendrites with some containing synaptic vesicles (DCSV). Ten types of axon terminal were identified and divided into two categories. The first, GABA-immunonegative and making asymmetrical synaptic contact, contain round (RT1, RT2, RT3) or pleiomorphic synaptic and many dense-core vesicles (DCT). RT1 terminals are retinothalamic and RT2 terminals hyperpalliothalamic; both mainly contact dendrites of projection neurons (72% and 78% respectively), less frequently dendrites of interneurons and sometimes DCSV; RT1 terminals are rarely involved in synaptic triads. The second category are consistently GABA-immunopositive. Four types (PT1-4), distinguished by their pleiomorphic synaptic vesicles, make symmetrical synaptic contact essentially with dendrites of projection neurons, more rarely on dendrites of interneurons (PT2). PT1 terminals are very probably those of interneurons, whereas the rare PT4 terminals are of retinal origin. A fifth type (RgT) contains round synaptic vesicles and makes asymmetrical synaptic contact with dendrites of projection neurons and interneurons. PT2 and RgT terminals occasionally contact DCSV of interneurons, which are sometimes involved in synaptic triads. Two final subcategories (DCgT1-2) contain many dense-core vesicles. Our findings are compared with those of previous studies concerning the fine structure and neurochemical properties of the GLd of reptiles and mammals, with special reference to the origin of the extraretinal and extracortical projections to this structure.

Tectorotundal connections in turtles: an electron microscopic tracing and GABA-immunocytochemical study.

The nucleus rotundus of the turtles Emys orbicularis and Testudo horsfieldi was analysed by axonal tracing methods and post-embedding GABA immunocytochemistry. After injections of horseradish peroxidase or biotinylated dextran amine into the optic tectum, electron microscopic observations showed that the vast majority of ipsilateral tectorotundal axon terminals were small in size, had smooth contours and contained small, round, densely packed synaptic vesicles. These terminals were GABA-immunonegative, often gathered in clusters, and established asymmetrical synaptic contacts with either small- or medium-sized GABA-negative dendritic profiles and with GABA-immunoreactive (GABA-ir) dendrites, which did not contain synaptic vesicles. Occasional GABA-ir-labelled axon terminals were observed; these may arise from the rare GABAergic neurons in the central tectal layer, or from neurons in the ventral pretectal nucleus, which projects both to the optic tectum and nucleus rotundus. In addition to tracer-labelled axon terminals, we observed both GABA-negative and GABA-ir cell bodies and dendrites also labelled by the tracer. No GABA-ir presynaptic dendritic profiles containing synaptic vesicles were observed. The existence in reptiles of reciprocal connections between the nucleus rotundus and the optic tectum as a phylogenetically ancient feedback system is discussed.