Relative number of cells projecting from contralateral and ipsilateral nucleus isthmi to loci in the optic tectum is dependent on visuotopic location: horseradish peroxidase study in the leopard frog.

The leopard frog optic tectum is the principal target of the contralateral retina. The retinal terminals form a topographic map of the visual field. The tectum also receives bilateral topographic input from a midbrain structure called nucleus isthmi. In this study we determined the relative strength of n. isthmi projections to different loci in the tectum. Horseradish peroxidase (HRP) was applied at single superficial tectal locations in a series of leopard frogs. The application sites were distributed across the tectum. Retrogradely filled cells were counted in ipsilateral and contralateral nucleus isthmi. Although all regions of the tectum receive input from both n. isthmi, the relative number of labeled cells in the two n. isthmi is dependent on visuotopic location. Input to the rostromedial tectum representing the visual field ipsilateral to the labeled tectum comes primarily from the contralateral n. isthmi. Input to the caudolateral tectum representing the visual field contralateral to the labeled tectum originates mostly from the ipsilateral n. isthmi. Tectal application sites representing the visual midline had approximately equal numbers of labeled cells in the two n. isthmi. The results are similar at postapplication survival times ranging from 2 to 14 days. Using application of HRP to rostral tectum and application of nuclear yellow to caudal tectum, we show that the anisotropy in isthmi labeling is not due to take up of these labels by isthmotectal fibers passing through the application sites that terminate elsewhere.     

Relationship between isthmotectal fibers and other tectopetal systems in the leopard frog

We studied the relationship of isthmotectal input to other tectal afferent fiber systems in three ways. 1) Using horseradish peroxidase (HRP) histochemistry, we determined the nonretinal inputs to the superficial tectum. In different sets of animals we a) applied HRP to the tectal surface; b) inserted HRP crystals into the tectum; c) injected small volumes of HRP solutions into the superficial tectum. N. isthmi accounts for more than 65% of the nonretinal extrinsic input in the superficial tectal layers. One set of fibers from the contralateral n. isthmi projects to the most superficial layer. Fibers from posterior thalamus and tegmentum project to both superficial and deeper layers in the tectum, but not to the most superficial layer. The ipsilaterally projecting isthmotectal fibers terminate in the deeper superficial layers. 2) We investigated the relationship between retinofugal and contralaterally projecting isthmotectal pathways. We orthogradely labelled n. isthmi fibers by unilateral HRP injections into n. isthmi, and we also labelled retinal fibers by injecting tritiated l-proline into both eyes. In such animals contralaterally projecting isthmotectal fibers cross in the dorsal posterior region of the optic chiasm. From the chiasm to the tectum isthmotectal fibers and retinofugal fibers are admixed. 3) We determined whether other fiber systems cross with contralaterally projecting isthmotectal fibers. We cut the posterior part of the optic chiasm and applied HRP crystals to the cut. Only n. isthmi and retina are retrogradely labelled.     

Fiber connections of the nucleus isthmi in the carp (Cyprinus carpio) and tilapia (Oreochromis niloticus)

The nucleus pretectalis (NP) is a prominent nucleus in the percomorph pretectum and has been shown to project to the nucleus isthmi in the filefish by an HRP tract-tracing method [Ito et al., 1981], but a homologous nucleus to the NP is apparently lacking in ostariophysans. The present study examined fiber connections of the nucleus isthmi in an ostariophysan teleost, the carp (Cyprinidae, Cyprinus carpio), to identify a nucleus homologous to the percomorph nucleus pretectalis. Identical studies in a percomorph tilapia (Cichlidae, Oreochromis niloticus) were also performed. Injections of biotinylated dextran amine (BDA) or biocytin to the carp nucleus isthmi labeled cells in the ipsilateral optic tectum and nucleus ruber of Goldstein [1905]. Labeled tectal neurons were located in the stratum periventriculare (SPV) and the stratum fibrosum et griseum superficiale (SFGS). The somata in the SPV were pyriform and those in the SFGS were fusiform. No labeled cells were found in the pretectum. Labeled terminals were seen in the ipsilateral nucleus pretectalis superficialis pars parvocellularis (PSp), optic tectum, and bilateral nucleus ruber. Terminals in the nucleus ruber appear to come from tectal neurons in the SFGS labeled by isthmic injections. Thus the nucleus isthmi has reciprocal fiber connections with the ipsilateral optic tectum, receives projections from the ipsilateral nucleus ruber, and projects to the ipsilateral PSp. The nucleus pretectalis homologue is apparently absent in the carp. Studies in tilapia showed that the nucleus isthmi receives bilateral projections from the NP and optic tectum. In addition, the present study revealed a previously unknown afferent from the nucleus ruber to the percomorph nucleus isthmi. The tilapia nucleus isthmi projects to the same targets as in the carp. Isthmic projection neurons in the tilapia optic tectum were located in the SPV and pyriform with a similar shape to the carp SPV neurons that project to the nucleus isthmi. No labeled cells were found in the SFGS of tilapia optic tectum. The fusiform neurons in the SFGS of the carp optic tectum possess various hodological similarities with the NP and may correspond to the NP neurons of percomorphs.     

Localization of neurons afferent to the optic tectum in longnose gars

Afferent pathways to the optic tectum in the longnose gar were determined by unilateral tectal injections of HRP. Retrogradely labeled cells were observed in the ipsilateral caudal portion of the rostral entopeduncular nucleus and bilaterally in the rostral half of the lateral zone of area dorsalis of the telencephalon. The following diencephalic cell groups were also labeled following tectal injections: the ipsilateral anterior, ventrolateral, and ventromedial thalamic nuclei, the periventricular pretectal nucleus, and the central pretectal nucleus (bilaterally); the ventromedial thalamic and central pretectal nuclei revealed the largest number of labeled cells. At midbrain levels, retrogradely labeled cells were seen in the ipsilateral torus longitudinalis, nucleus isthmi, and accessory optic nucleus; cells were labeled bilaterally in the torus semicircularis and a rostral tegmental nucleus. Only a few cells were labeled in the contralateral optic tectum, suggesting that few of the fibers of the intertectal commissure are actually commissural to the tectum. At hindbrain levels, retrogradely labeled cells were seen bilaterally in the locus coeruleus, the superior, medial, and inferior reticular formations, the eurydendroid cells of the cerebellum, and the nucleus of the descending trigeminal tract; the contralateral dorsal funicular nucleus also exhibited labeling. Clearly, the tectum in gars receives a substantial number of nonvisual afferents from all major brain areas, most of which have been reported in other vertebrates. The functional significance of these afferent sources and their probable homologues in other vertebrate groups are discussed.     

Anatomy and physiology of a binocular system in the frograna pipiens

The locations of tectal neurons projecting to nucleus isthmi (n. isthmi) were found by iontophoretic injection of horseradish peroxidase (HRP) into n. isthmi. After retrograde transport, stained tectal somata are found to lie almost exclusively in layer 6 and below of the ipsilateral tectum. Many cells are colored throughout the extent of their dendrites into the fine rami, giving the appearance of a Golgi stain. Nucleus isthmi receives projections from the ipsilateral tectum and from no other region. Nucleus isthmi units recorded electrically respond to visual stimuli and are arranged in a topographic map of the visual field. There are two types of receptive fields, those with small centers and those with large centers. The small centers are about 3-5 degrees in diameter, similar to type 2 optic nerve fibers. Their response is to many of the same geometric features of stimulus as excite type 2 fibers. The large centers are at least 7-10 degrees in diameter and respond to many of the same features as excite types 3 and 4 optic nerve fibers. The responsiveness of small and large center n. isthmi units is very similar to the elements of the ipsilateral visual field projection onto tectum, i.e. the neuropilar units recorded in layers A and 8 of the tectum when the contralateral eye is occluded. These are in strong contrast to those of tectal cells of layer 6 and below, which have large receptive fields, show far less vivacious response, adapt extremely rapidly to repeated stimuli and are hard to describe in terms of characteristic stimuli because they are unresponsive most of the time. We suggest, therefore, that the axons of tecto-isthmic cells are quite active and that their cell bodies, located in layer 6 and below, only fire occasionally on the firing of their axons.     

Topographic projections between the nucleus isthmi and the optic tectum in a teleost,Navodon Modestus

Following horseradish peroxidase injections into the optic tectum of a teleost,Navodon modestus, reciprocal and topographic projections between the nucleus isthmi and the ipsilateral optic tectum were determined. The isthmo-tectal fibers diverge to the optic tectum while maintaining the spatial arrangements of the isthmic cells from which the fibers originate. The tecto-isthmic projections also keep the spatial arrangements in the optic tectum. The tectal fibers converge near the nucleus isthmi and terminate in the non-cellular portion of the nucleus. The reciprocal topography is apparent in the combined results of 9 experiments with one tectal injection in each region. No labeled cells and fibers were found in the contralateral nucleus isthmi.     

Isthmotectal axons make ectopic synapses in monocular regions of the tectum in developing Xenopus laevis frogs.

During the development of binocular maps in the tectum of Xenopus laevis, axons that relay input from the ipsilateral eye via the nucleus isthmi undergo a prolonged period of shifting connections. This shifting accompanies the dramatic change in eye position that takes place as the laterally placed eyes of the tadpole move dorsofrontally. There is a concomitant expansion of the proportion of tectum that receives contralateral retinotectal input corresponding to the binocular portion of the visual field. Electrophysiological recording demonstrates that ipsilateral units are present in those rostral tectal zones, and anatomical methods show that the isthmotectal axons arborize densely in the rostral region but also extend sparser branches into the caudal zone, which is occupied by contralateral inputs with receptive fields in the monocular zone of the visual field. A mechanism that aligns the ipsilateral and contralateral maps is activity-dependent stabilization of isthmotectal axons that exhibit firing patterns correlated with those of nearby retinotectal axons. In order for activity patterns to function in stabilizing correct connections and promoting the withdrawal of incorrect connections, synaptic communication of some sort is hypothesized to be essential. We have investigated whether isthmotectal axons make morphologically identifiable synapses during development and where such synapses are located. We find evidence for morphologically identifiable synapses in all regions of the tectum, along with many growth cones and structures that are probably immature synapses. As in the adult, the synapses contain round, clear vesicles, have asymmetric specializations, and terminate on structures that appear to be dendrites. In both adult and tadpole, the rarity of serial synapses involving isthmotectal terminals suggests that the interactions between retinotectal and isthmotectal inputs are mediated by postsynaptic dendrites.     

Cytoarchitecture,fiber connections,and ultrastructure of the nucleus pretectalis superficialis pars magnocellularis (PSm) in carp

The cytoarchitecture, fiber connections, and ultrastructure of the nucleus pretectalis superficialis pars magnocellularis (PSm) were studied in cypriniform teleosts (Cyprinus carpio). The PSm is an oval nucleus in the pretectum. Medium-sized cells and synaptic glomeruli are the main components of the nucleus. A lesser number of small cells are also present. Most of the medium-sized cells form one or two cell layers on the periphery of the nucleus, and some cells are scattered among synaptic glomeruli in the nucleus. Cell bodies in the peripheral cell layer are pyriform and sprout a thick dendrite directed inward. The dendrite gives off fine dendritic branches, which are postsynaptic elements in synaptic glomeruli. The PSm projects to the ipsilateral corpus mamillare (CM) and sends collaterals to the ipsilateral nucleus lateralis valvulae (NLV). Axons of the PSm neurons have terminals with many varicosities in the CM, and collaterals in the NLV have cup-shaped terminals around the cell bodies of the NLV neurons. Following horseradish peroxidase (HRP) injections into the PSm, HRP-labeled cells are found ipsilaterally in the optic tectum, the nucleus tractus rotundus of Schnitzlein, and the nucleus ruber of Goldstein. The tecto-PSm projections are topographically organized. The rostral optic tectum projects mainly to the rostral portion of the PSm, and the caudal tectum projects to the caudal portion of the PSm. The ventral tectum sends fibers mainly to the ventral part of the PSm. The dorsomedial tectum projects to the medial part of the PSm, and the dorsolateral tectum projects to the lateral part of the PSm. Tectal projection neurons to the PSm are of only one type. The tectal cell body is pyriform and is situated in the superficial part of the ipsilateral stratum periventriculare (SPV). The tectal neurons have a long perpendicular dendrite, which branches out in the stratum opticum (SO). An axon emerges from the branching site in the SO. Judging from the dendritic branching pattern of the tectal projection neurons, we concluded that the PSm receives visual information from the optic tectum. © 1993 Wiley-Liss, Inc.     

Ultrastructure of the crossed isthmotectal projection in Xenopus frogs

The nucleus isthmi NI of frogs is a relay for input from the eye to the ipsilateral tectum; each NI receives retinotopic input from one tectum and sends retinotopic output to both tecta. The crossed isthmotectal projection in Xenopus displays tremendous plasticity during development. Physiological and anatomical studies have suggested that the location at which a developing isthmotectal axon will terminate is determined by the correlation of its visually evoked activity with the activity of nearby retinotectal terminals. What structures could mediate such communication? We have examined quantitatively the ultrastructural characteristics of crossed isthmotectal axons and synapses in order to determine whether retinotectal axons communicate directly with isthmotectal axons via axo-axonic synapses or whether the communication is indirect, e.g., via common postsynaptic dendrites. Our results support the conclusion that isthmotectal axons interact with retinotec tal axons indirectly and that tectal cell dendrites are the critical site of interaction.     

Caudal topographic nucleus isthmi and the rostral nontopographic nucleus isthmi in the turtle, Pseudemys scripta

Isthmotectal projections in turtles were examined by making serial section reconstructions of axonal and dendritic arborizations that were anterogradely or retrogradely filled with HRP. Two prominent tectal-recipient isthmic nuclei--the caudal magnocellular nucleus isthmi (Imc) and the rostral magnocellular nucleus isthmi (Imr)--exhibited strikingly different patterns of organization. Imc cells have flattened, bipolar dendritic fields that cover a few percent of the area of the cell plate constituting the nucleus and they project topographically to the ipsilateral tectum without local axon branches. The topography was examined explicitly at the single-cell level by using cases with two injections at widely separated tectal loci. Each Imc axon terminates as a compact swarm of several thousand boutons placed mainly in the upper central gray and superficial gray layers. One Imc terminal spans less that 1% of the tectal surface. Imr cells, by contrast, have large, sparsely branched dendritic fields overlapped by local axon collaterals while distally, their axons nontopographically innervate not only the deeper layers of the ipsilateral tectum but also ipsilateral Imc. Imr receives a nontopographic tectal input that contrasts with the topographic tectal input to Imc. Previous work on nucleus isthmi emphasized the role of the contralateral isthmotectal projection (which originates from a third isthmic nucleus in turtles) in mediating binocular interactions in the tectum. The present results on the two different but overlapping ipsilateral tecto-isthmo-tectal circuits set up by Imc and Imr are discussed in the light of physiological evidence for selective attention effects and local-global interactions in the tectum.     

Topographic and morphometric effects of bilateral embryonic eye removal on the optic tectum and nucleus isthmus of the leopard frog

Rana pipiens were raised through metamorphosis after extirpation of both eye primordia at Shumway embryonic stage 17 (Shumway '40). The visual connections between the isthmic nuclei and the optic tectum were examined in these animals using horseradish peroxidase (HRP) histochemistry. Isthmo-tectal projections are normally aligned with the primary retinotectal map. We asked whether these connections would develop normal topographic organization in the absence of normal retinal input. HRP was formed into a solid pellet (? 200–500 μm diameter) and inserted into one tectal lobe on the tip of a fine metal probe. The procedure produced relatively restricted retrograde label in somas and dendrites in both isthmi nuclei. In the nucleus isthmus ipsilateral to the tectal lobe receiving the HRP pellet, processes of tecto-isthmi neurons were labeled by anterograde transport. The topography of the isthmo-tectal and tecto-isthmic projections were identical in the developmentally enucleated animals and in normal frogs, even though eye removal severely reduced the volume of the optic tecta and the isthmi nuclei. Thus our analyses indicate that retinal contacts do not play an active role in the development of the positional or polarity cues that are involved in “mapping” projections between central visual nuclei. These results are discussed in the context of peripheral specification of central connections and in terms of models that have recently been proposed to explain the development of the retinotectal system.     

Cytoarchitecture,fiber connections,and ultrastructure of nucleus isthmi in a teleost (Navodon modestus) with a special reference to degenerating isthmic afferents from optic tectum and nucleus pretectalis

Cytoarchitecture and fiber connections of the nucleus isthmi in a teleost (Navodon modestus) were studied by means of Nissl, Bodian, toluidine blue, Golgi, and Fink-Heimer methods. Synaptic terminals were classified by the ultrastructural characteristics, and their origins were determined by electron microscopic degeneration experiments. The nucleus isthmi is composed of an outer cellular area or shell and an inner noncellular area or core. The shell covers anterior, dorsal, and ventral aspects of the core. The cell bodies in the shell are oval (15 × 20 μm) with an anteroposterior long axis, and have many somatic spines. Spines are also seen on the initial segment of the axon. Primary dendrites extend postermedially and branch out in the core. The core contains thin and thick myelinated fibers, which originate in the optic tectun and in the nucleus pretectalis, respectively. At least two types of axons terminal were distinguished in the nucleus isthmi: S type, containing spherical vesciles, and F type, containing flattened vesicles. S terminals are derived from thin myelinated fibers and are only seen in the core where they form asymmetric synapses with dendrites. Frequently a portion of the S terminal membrane near the usual synaptic cleft is in close apposition with the membrane of an adjacent small dendrite or spine. F terminals, which derived from thick myelinated fibers, make symmetric synaptic contacts with both cell bodies in the shell and dendrites in the core. S terminals degenerate after ipsilateral ablation of the optic tectum, whereas F terminals degenerate after destruction of the nucleus pretectalis.     

Identification of a nucleus isthmi in the weakly electric fish Apteronotus leptorhynchus (Gymnotiformes)

The nucleus isthmi of teleost fish, amphibians, reptiles and birds, and its probable homologue, the nucleus parabigeminalis of mammals, share in common certain features such as location in the dorsal tegmentum and reciprocal connectivity with the optic tectum. In gymnotid fish the nucleus isthmi is located dorsolaterally in the brainstem tegmentum, ventral to the torus semicircularis and the lateral mesencephalic reticular area and dorsal to the rostral nucleus praeeminentialis. The nucleus isthmi has an ovoid shape, with a compact cellular part on its dorsal, medial and ventral aspects surrounding a hilar region with a sparse population of larger cells. Following wheat germ agglutinin-conjugated horseradish peroxidase injections into the optic tectum, anterogradely labeled fine terminals were observed leaving the tectobulbar tract and entering the ipsilateral nucleus isthmi via its laterally facing hilar region. Retrogradely labeled cells were present in the nucleus isthmi on both sides, indicating the presence of a bilateral isthmotectal projection similar to that reported in amphibians. The putative isthmal nucleus stains densely for acetylcholinesterase. Based on the similarity of its location, shape, cholinesterase histochemistry and reciprocal connectivity with the optic tectum, we identified this structure as the nucleus isthmi of gymnotids. An interesting observation of this study was that the nucleus isthmi, in addition to receiving fine terminals from the optic tectum, is also the recipient of a sparser population of thicker-caliber afferent fibers which terminate not only in the large-celled hilar region but also within the smaller-celled component of the nucleus; this projection appears to emanate from the torus semicircularis dorsalis.     

Topographic projections between the nucleus isthmi and the tectum of the frog Rana pipiens.

The connections between the nucleus isthmi and the tectum in the frog have been determined by several anatomical techniques: iontophoresis of horseradish peroxidase into the tectum, iontophoresis of 3H-porline into the nucleus isthmi and the tectum, and Fink-Heimer degeneration staining after lesions of the nucleus isthmi. The results show that the nucleus isthmi projects bilaterally to the tectal lobes. The ipsilateral isthmio-tectal fibers are distributed in the superficial layers of the tectum, coincident with the retionotectal terminals. The contralateral isthmio-tectal fibers travel anteriorly adjacent to the lateral optic tract and cross the midline in the supraoptic ventral decussation, where they turn dorsally and caudally; upon reaching the tectum, the fibers end in two discrete layers, layers 8 and A of Potter. The tectum projects to the ipsilateral nucleus isthmi and there is a reciprocal topographic relationship between the two structures. Thus, a retino-tecto-isthmio-tectal route exists which may contribute to the indirect ipsilateral retinotectal projection which is observed electrophysiologically. The connections between the nucleus isthmi and the tectum in the frog are strinkingly similar to the connections between the parabigeminal nucleus and the superior colliculus of mammals.     

Isthmic afferent neurons identified by the retrograde HRP method in a teleost, Navodon modestus

Isthmic afferent neurons were investigated by the retrograde horseradish peroxidase (HRP) method in a teleost, Navodon modestus. Following HRP injections into the nucleus isthmi, large pyriform neurons are labeled in the ipsilateral optic tectum. Very large and multipolar neurons are also labeled in the ipsilateral nucleus pretectalis. No labeled neurons were found in other areas.     

Cytoarchitecture, synaptic organization and Fiber Connections of the nucleus olfactoretinalis in a teleost (Navodon modestus)

Cytoarchitecture, synaptic organization and fiber connections of the nucleus olfactoretinalis (NOR) in a teleost, Navodon modestus, have been studied light- and electron-microscopically using an HRP or HRP-degeneration combined method. Following HRP injections into the optic nerve, most contralateral and a few ipsilateral neurons in the NOR were labeled. There are two types of neurons in NOR. Type I neurons have a medium-sized spindle-shaped soma with a round nucleus, and type II neurons have a large oval soma with an invaginated nucleus and contain cored vesicles (80-130 nm in diameter). Afferent terminals which form synaptic contacts with cell bodies of NOR neurons were classified into 3 types according to their morphological characteristics; S, F1 and F2 terminals. S terminals originated in ipsilateral area ventralis telencephali pars supracommissuralis (Vs). These terminals contain both spherical and cored vesicles, and make synaptic contacts with both type I and type II neurons. F1 terminals, which originated in ipsilateral area dorsalis telencephali pars posterior (Dp), are large in profile, and contain flat vesicles and mitochondria with irregularly arranged cristae. These terminals make synaptic contacts only with type I neurons. F2 terminals are small in profile, and contain flat vesicles, cored vesicles and small mitochondria with regularly arranged cristae. F2 terminals make synaptic contacts with both type I and type II neurons. The functional significance of NOR and the relationship between NOR and the ganglion of the nervus terminalis are discussed.     

Direct synaptic projections to esophageal motoneurons in the nucleus ambiguus from the nucleus of the solitary tract of the rat

Neurons of the nucleus of the solitary tract (NTS) serve as interneurons in swallowing. We investigated the synaptology of the terminals of these neurons and whether they project directly to the esophageal motoneurons in the compact formation of the nucleus ambiguus (AmC). Following wheat germ agglutinin conjugated horseradish peroxidase (WGA-HRP) injection into the NTS, many anterogradely labeled axodendritic terminals were found in the neuropil of the AmC. The majority of labeled axodendritic terminals (89%) contained round vesicles and made asymmetric synaptic contacts (Gray's type I), but a few (11%) contained pleomorphic vesicles and made symmetric synaptic contacts (Gray's type II). More than half of the labeled terminals contacted intermediate dendrites (1-2 μm diameter). There were no retrogradely labeled medium-sized motoneurons, but there were many retrogradely labeled small neurons having anterogradely labeled axosomatic terminals. A combined retrograde and anterograde transport technique was developed to verify the direct projection from the NTS to the esophageal motoneurons. After the esophageal motoneurons were retrogradely labeled by cholera toxin subunit B conjugated HRP, the injection of WGA-HRP into the NTS permitted ultrastructural recognition of anterogradely labeled axosomatic terminals contacting directly labeled esophageal motoneurons. Serial sections showed that less than 20% of the axosomatic terminals were labeled in the esophageal motoneurons. They were mostly Gray's type I, but a few were Gray's type II. In the small neurons, more than 30% of axosomatic terminals were labeled, which were exclusively Gray's type I. These results indicate that NTS neurons project directly not only to the esophageal motoneurons, but also to the small neurons which have bidirectional connections with the NTS. J. Comp. Neurol. 381:18-30, 1997. © 1997 Wiley-Liss, Inc.     

Electron microscopic study of the rubrocerebellar projection in the cat

Rubral neurons sending axons to the cerebellar anterior interpositus nucleus (AIN) in the cat were identified light microscopically by labeling them with horseradish peroxidase (HRP). The synaptic organization of these rubral neurons and of their afferents from the cerebral motor cortex and the AIN was also analyzed electron microscopically by combined anterograde degeneration and retrograde HRP-labeling techniques. In the light microscopic study, either HRP or a mixture of HRP and kainic acid was injected into the AIN. Both of the injections resulted in retrograde labeling of rubrocerebellar projection neurons in the red nucleus on the contralateral side. The labeled neurons were distributed throughout the rostrocaudal extent of the red nucleus: some lay in clusters. Most labeled neurons were small to medium-sized, although some were large. The injection of HRP into the AIN also resulted in anterograde labeling of cerebellorubral projection fibers terminating in a wider area of the red nucleus on the contralateral side of the injection, whereas the injection of a mixture of HRP and kainic acid showed no anterograde labeling of fibers or terminals. In one set of electron microscopic observations, HRP injections into the AIN were combined with ablation of the motor cortex. Degenerating axon terminals were occasionally found to synapse with both dendrites and neuronal somata labeled with HRP retrogradely. In another set of electron microscopic observations, a mixture of HRP and kainic acid was injected into the AIN in order to label rubrocerebellar projection neurons retrogradely and to bring about degeneration in the cerebellorubral projection fibers anterogradely. Abundant degenerating axon terminals were observed to make axosomatic synaptic contacts with rubral neurons labeled with HRP retrogradely and also with unlabeled rubral neurons. These results indicate that cerebrorubrocerebellar and rubrocerebellorubral monosynaptic circuitries exist which constitute one of the cerebrocerebellar linkages, as well as those linkages via the inferior olivary complex and the pontine nuclei.     

Nucleus isthmi enhances calcium influx into optic nerve fiber terminals in Rana pipiens

We examined the role of nucleus isthmi in enhancing intracellular calcium concentrations in retinotectal fibers in the frog optic tectum in vitro. The intracellular calcium levels were measured using the fluorescent calcium-sensitive dye, Calcium Green-1 3000 mw dextran conjugate (CG-1), which was injected into one optic nerve. Electrical stimulation of the labeled optic nerve alone increased tectal CG-1 fluorescence whereas electrical stimulation of nucleus isthmi alone had no effect on CG-1 fluorescence. Electrical stimulation of the nucleus isthmi ipsilateral to the labeled tectum, followed by electrical stimulation to the optic nerve can enhance calcium uptake more than a double pulse stimulation of the optic nerve alone. Maximum enhancement of the calcium signal by nucleus isthmi occurs when optic nerve stimulation follows the ipsilateral nucleus isthmi stimulation by 10 ms. These results suggest that nucleus isthmi input can facilitate retinotectal neurotransmission, and the mechanism could be used to allow the frog to attend to a single prey stimulus in an environment of several prey stimuli.     

Distribution, laminar location, and morphology of tectal neurons projecting to the isthmo-optic nucleus and the nucleus isthmi, pars parvocellularis in the pigeon (Columba livia) and chick (Gallus domesticus): a retrograde labelling study.

Retrograde transport of Phaseolus vulgaris leucoagglutinin (PHA-L), fluorogold, fast blue, rhodamine labelled microspheres, and horseradish peroxidase (HRP) was employed to study the distribution, laminar location within the optic tectum, and morphology of tectal cells projecting upon the isthmo-optic nucleus (ION) and the nucleus isthmi, pars parvocellularis (Ipc), in the pigeon and chick. Following injections into the ION, all retrograde markers labelled tecto-ION neurons and their dendrites in the ipsilateral tectum. The cells were located within a relatively narrow band at the border between layers 9 and 10 of the stratum griseum et fibrosum superficiale (SGFS). Retrogradely labelled neuronal somata were different in both dendritic branching and shape; however, tecto-ION neurons generally possessed non-spiny radially oriented and multi-branched dendrites. The apical processes extended into the retino-recipient layers (2-7) of the SGFS and basal dendrites extended into layers 12-14 of the SGFS. Positive neuronal somata were observed throughout the rostro-caudal extent of the optic tectum. The average distance between adjacent tecto-ION neurons varied from one region to another. Specifically, retrogradely labelled cells were more numerous in the caudal, lateral, and ventral tectum, and less numerous at rostro-dorsal levels. Approximately 12,000 tecto-ION neurons were labelled within the ipsilateral optic tectum following either PHA-L or fluorescent dye injections. While the regional distribution of tecto-Ipc neurons was not examined, the morphology indicated that the cells had a single radially oriented dendritic process. Therefore, the apical dendrites are more restricted than those of tecto-ION cells. Moreover, the dendrites were spiny and arborized within layers 3, 5, and 9 of the ipsilateral optic tectum. The axon of tecto-Ipc cells arise from the apical process as a shepherd's crook and descend into the deep layers of the optic tectum. These results indicate that 1) tecto-ION and tecto-Ipc neurons are possibly monosynaptically activated by retinal input, 2) tecto-ION neurons are heterogeneous in morphology, and 3) there is a differential distribution of the tecto-ION neurons throughout the rostro-caudal extent of the optic tectum, suggesting a greater representation of the caudo-ventral portion of the optic tectum within the ION. The discussion primarily concerns the organization of the retino-tecto-ION-retinal circuit in light of the distribution and morphology of tecto-ION neurons within the optic tectum.