Isthmotectal Projection Research Articles

Current source density analysis of contra- and ipsilateral isthmotectal connections of the frog

The nucleus isthmi (NI) of the frog receives input from the ipsilateral optic tectum and projects back to both optic tecta. After ablation of NI, frogs display no visually elicited prey-catching or threat avoidance behavior. Neural mechanisms that underlie the loss of such important behavior have not been solved. Electrophysiological examination of the contralateral isthmotectal projection has proved that it contributes to binocular vision. On the other hand, there are very few physiological investigations of the ipsilateral isthmotectal projection. In this study, current source density (CSD) analysis was applied to contra- and ipsilateral isthmotectal projections. The contralateral projection produced monosynaptic sinks in superficial layers and in layer 8. The results confirmed former findings obtained by single unit recordings. The ipsilateral projection elicited a prominent monosynaptic sink in layer 8. Recipient neurons were located in layers 6-7. These results, combined with those from the former intracellular study, led to the following neuronal circuit. Afferents from the ipsilateral NI inhibit non-efferent pear shaped neurons in the superficial layers, and strongly excite large ganglionic neurons projecting to the descending motor regions. Thus feedback to the output neurons strengthens the visually elicited responses.

Effects of choline and other nicotinic agonists on the tectum of juvenile and adult Xenopus frogs: a patch-clamp study

We have used anatomical methods and whole-cell patch-clamp recording to assess the distribution of nicotinic receptors in the tectum of Xenopus frogs and to measure effects of nicotinic ligands (carbachol, cytisine and nicotine) on glutamatergic spontaneous miniature excitatory postsynaptic currents. Our results confirm that retinotectal axons account for the majority of nicotinic receptors in the tectum and that nicotinic agonists exert presynaptic effects that increase the rate of transmitter release on to tectal cells. The nicotinic blockers mecamylamine and methyllycaconitine reduced responses to carbachol and cytisine. A small percentage of cells also showed postsynaptic responses. We have assessed whether there are developmental changes in the frequency of occurrence of spontaneous miniature excitatory postsynaptic currents. The first three months post-metamorphosis fall within the critical period for the dramatic plasticity displayed by binocular inputs during development in Xenopus. During this period, visual activity governs the formation of orderly maps relayed from the ipsilateral eye via the cholinergic projection from the nucleus isthmi to the tectum. In this study, we have found that critical-period tecta (two to 12 weeks postmetamorphosis) tend to have higher spontaneous activity than do older tecta (two to 69 weeks postmetamorphosis), and that nicotinic agonists increase that activity in both groups, with the result that the peak rates in response to nicotinic agonists are higher during the critical period than later. We also investigated the possible role of choline as an agonist of nicotinic receptors in the tectum. We have found that choline, as well as carbachol and cytisine, can cause a reversible increase in the rate of miniature excitatory postsynaptic currents. This result may help to explain how the isthmotectal projection, which accounts for the overwhelming majority of cholinergic input to the tectum, can exert effects on retinotectal terminals even though there are no morphologically identifiable synapses between the two populations. We have examined the morphology of cells filled with biocytin during the patch-clamp experiments, and we find that cells with dendrites in the stratum zonale, a layer with particularly dense input from the contralateral nucleus isthmi, have higher spontaneous activity than cells with dendrites that do not extend into that layer. Nicotinic agonists increased the activity recorded in both classes of cells. In addition, four pretectal cells were identified. Nicotinic agonists increased the rate of spontaneous activity recorded in that population. The results indicate that retinotectal transmission in the superior colliculus can be increased presynaptically by activity of the cholinergic projections of the nucleus isthmi. This modulation may be the basis for observations that blocking of cholinergic input disrupts the formation of topographic retinotectal projections. Moreover, the ability of choline to activate these receptors suggests that this metabolite of acetylcholine may permit paracrine activation of presynaptic receptors even though the tectum contains high acetylcholinesterase activity.

Seeing beyond the midline: the role of the contralateral isthmotectal projection in the leopard frog.

The ground level visual field of each eye of the leopard frog includes the entire ipsilateral 180-deg field and approximately 60 deg into the frontal contralateral field. When one eye is covered with an opaque patch, a frog responds to prey stimuli over the entire field of the other eye. Nevertheless, when one optic nerve is cut, the animal responds to prey in the ipsilateral hemifield of the connected eye, but only responds as far as about 30 deg past the frontal midline. If one optic tract is cut, the animal does not respond to prey past the frontal midline. We hypothesized that the responses past the frontal midline might be mediated by input from contralaterally projecting isthmotectal fibers. These fibers originate in the nucleus isthmi, a posterior midbrain structure. We found that when we placed an opaque patch over one eye and either ablated the ipsilateral nucleus isthmi, or cut crossing isthmotectal fibers in the optic chiasm, or blocked input to nucleus isthmi by ablating the ipsilateral tectal lobe, animals did not respond to prey stimuli past the frontal midline. We found that when we placed an opaque patch over one eye and cut crossing optic fibers in the anterior part of the optic chiasm (sparing crossing isthmotectal fibers), animals responded to prey stimuli in the nasal half of the seeing eye's contralateral frontal field. Our results suggest that contralaterally projecting isthmotectal fibers enable the frog to respond to stimuli past the frontal midline. We suggest a one-dimensional model of how nucleus isthmi influences tectal function.

Experience-dependent mechanism of binocular map plasticity in Xenopus: incongruent connections are masked by retinal input

When one eye in larval Xenopus is rotated to change the orientation of its visual map on the contralateral tectum, the map from the other eye to the same tectum can subsequently alter its orientation to match the rotated input. This plasticity re-establishes binocular congruence between the two maps and involves an experience-dependent reorganization of functional connections in the crossed isthmotectal projection, which delivers the ipsilateral eye's map to the tectum. Here, recordings were made at the same tectal sites in 8 eye-rotated frogs immediately before and after removing the rotated retina. All had congruent binocular maps prior to enucleation, with matching receptive fields in the two eyes at each recording site. In 5 frogs, eye removal unmasked additional, incongruent receptive fields in the ipsilateral eye, recorded from previously undetectable crossed isthmotectal connections occupying their original positions in the tectum. This result suggests that the plasticity also involves functional suppression of binocularly-incongruent inputs from the ipsilateral eye by retinotectal afferents.

GABA as an inhibitory transmitter in the pigeon isthmo-tectal pathway

In the present investigation, the effects of inhibitory amino acids and their antagonists were tested on isthmo-tectal projection in the pigeon. The majority of superficial tectal cells are inhibited following stimulation of the parvocellular division of nucleus isthmi. A smaller portion of tectal cells showed excitatory responses followed by an inhibitory period. Both inhibitory mechanisms are blocked by the specific γ-aminobutyric acid (GABA)-antagonist bicuculline but not by strychnine. Our results support the idea that GABA acts as an inhibitory neurotransmitter in the pigeon isthmo-tectal pathways.

The development of non-retinal afferent projections to the frog optic tectum and the substance P immunoreactivity of tectal connections

Accessibility in early development and the presence of a retinotopic map have made the amphibian optic tectum a popular system for exploration of the role of synaptic function in central map refinement. Although a great deal is known about the development of retinal innervation of the tectum, little information exists about the development of non-retinal tectal inputs. Since these other afferent systems may contribute to the synaptic drive of developing tectal cells and thereby be involved in the activity-dependent refinement of the retinotectal map, we sought to determine whether these inputs are present at the early tadpole stages when the first retinal axons refine their synaptic order within the tectal neuropil. Rhodamine-labelled latex beads, retrogradely transported from injection sites in the optic tecta, were used to identify tectal afferent projections. Projection patterns in very young tadpoles were identical to those found in juvenile frogs and heavily labelled regions included areas of the posteroventral tegmental field, the posterior tuberculum, the ventromedial thalamic nucleus, the ventral part of the ventrolateral thalamic nucleus, the suprachiasmatic nucleus and discrete regions within the central and anterior thalamic nuclei. Ipsilateral nucleus isthmi cells were also labelled, indicating the existence of an isthmo-tectal projection in even the youngest animals examined. Additionally, substance P-like immunoreactive tecto-isthmal fibers were traced from the optic tectum to the nucleus isthmi. The presence of these connections suggests that feedback from the nucleus isthmi and/or input from the other brain areas projecting to the tectum may play a role in modulating the cellular mechanisms that underlie the formation of the visual map.

Xenopus exhibits seasonal variation in retinotectal latency but not tecto-isthmo-tectal latency.

1. The tectum of Xenopus receives visuotopic input from both eyes. The contralateral eye's projection reaches the tectum directly, via the optic nerve. The ipsilateral eye's projection reaches the tectum indirectly, via the nucleus isthmi and isthmo-tectal projection. 2. Because of the multi-synaptic nature of the ipsilateral pathway, there is an inherent delay between the time that information from the contralateral eye reaches the tectum and the time that information from the ipsilateral eye arrives at the tectum. The length of the intertectal delay is a function of the latencies of the contralateral and ipsilateral pathways. 3. The length of this intertectal delay has functional, as well as developmental, implications with regard to the role of N-methyl-D-aspartate receptors in tectal cell activity and development of orderly synaptic connections. 4. We have found that the latencies of the contralateral and ipsilateral pathways exhibit a seasonal variation, increasing during the winter months. The increases of both latencies during the winter were of similar magnitude, indicating that there were no significant changes in intertectal delay. The seasonal alteration in contralateral latency was not affected by dark-rearing and was affected to only a minor extent by a week-long alteration of ambient temperature.

Organization of ascending projections from the optic tectum and mesencephalic pretectal gray inRana pipiens

The ascending projections from the dorsal mesencephalon to the thalamus and pretectum in Rana pipiens were investigated by using the anterograde and retrograde transport of HRP with regard to two major issues: (1) the degree of tectotopic organization in the projections, and (2) their cells of origin. The results indicate that the spatial organization of the tecto-thalamic tract is specifically related to the laminar organization of the contributing tectal efferent neurons. Axons of neurons in the superficial portion of tectal layer 8 exit the tectum through layer 9 and travel in the superficial portion of the dorsal and ventral tecto-thalamic tracts and innervate the nucleus lentiformis mesencephali, the posterior lateral dorsal nucleus, and corpus geniculatum. The distribution of terminals within these structures varied with the tectal HRP-injection site. HRP injections in the ventral tecto-thalamic tract retrogradely labeled neurons in the superficial portion of tectal layer 8 across the lateral and caudal portion of the tectal lobe. HRP injections into the dorsal tecto-thalamic tract, at the level of the pretectum, retrogradely labeled pyriform neurons in the superficial portion of tectal layer 8 in the rostral and medial portions of the tectal lobe. With regard to the deep tectal layers, axons from pyramidal neurons in layer 6 and ganglionic neurons in layer 8 leave the tectum through layer 7, travel in both the dorsal and ventral tecto-thalamic tracts, and are located internal to the axons of the pyriform neurons of superficial tectal layer 8. The majority of the ganglionic neurons project to the posterior lateral ventral nucleus and the anterior lateral nucleus. The distribution of terminals within these nuclei did not display a tectotopic organization. A second major projection to the thalamus originates from the mesencephalic pretectal gray and innervates the nucleus lentiformis mesencephali, the posterior lateral dorsal nucleus, the anterior lateral nucleus, dorsal and ventral divisions of the ventral lateral thalamus, and the nucleus of Bellonci. Other axons from the mesencephalic pretectal gray terminate in the contralateral, medial portions of the posterior lateral dorsal thalamus, the ventral lateral thalamus, and the anterior lateral nucleus. The isthmo-tectal projection was also retrogradely labeled following tectal injections of HRP. This pathway travels in the most ventral portion of the ventral tecto-thalamic tract; its axons passed over the lateral margin of the endopeduncular nucleus bilaterally, and crossed the midline in the caudal portion of the optic chiasm. Extensive, bead-like varicosities were observed on these axons both in the endopeduncular nucleus and in the posterior optic chiasm.(ABSTRACT TRUNCATED AT 400 WORDS)

Anatomy, neurophysiology and functional aspects of the nucleus isthmi in salamanders of the family Plethodontidae

Tongue-projecting plethodontid salamanders have massive direct ipsilateral retinal afferents to the tectum opticum as well as a large and well developed nucleus isthmi. Retrograde staining revealed two subnuclei: A ventral one projecting to the contralateral tectal hemisphere and a dorsal one projecting back to the ipsilateral side. The isthmic nuclei show a retinotopic organization, which is in register with that of the tectum. Electrophysiological recordings from nucleus-isthmi neurons revealed response properties that are very similar to those found in tectal neurons. Thus, there is no substantial processing of tectal neural activity in the nucleus isthmi. Measurements of peak latencies after electrical and light stimulation suggest the continuous coexistence of 4 representations of the visual field in the tectum mediated by (1) the contralateral and (2) the ipsilateral direct retinal afferents, (3) the uncrossed and (4) the crossed isthmo-tectal projection. (1) and (2) originate at the same moment in the retina and arrive simultaneously in the tectum. It is assumed that in plethodontid salamanders with massive ipsilateral retino-tectal projections depth perception based on disparity cues is achieved by comparison of these images.

Chronic effects of NMDA and APV on tectal output in Xenopus laevis

In the South African clawed-toed frog Xenopus laevis, visual experience plays a crucial role in the formation of matching binocular maps in the tectum. The ipsilateral eye's projection, relayed through the crossed isthmotectal projection, displays marked plasticity in response to altered visual input during a critical period of development. This plasticity and the events responsible for the end of the critical period are mediated by N-methyl-D-aspartate (NMDA) receptor function. We have previously reported that chronic blockade of tectal NMDA receptors with the NMDA antagonist 5-amino-phosphonovaleric acid (APV) prevents plasticity of the crossed isthmotectal projection during the critical period, while chronic treatment with NMDA restores this plasticity after the end of the critical period. These results raise the question of whether the effects on plasticity are due to changes in electrical responsiveness of the treated tissue. In this study, we have quantitatively assessed the actions of APV and NMDA on certain aspects of tectal cell activity in Xenopus during and after the critical period by recording the output of the nucleus isthmi cells that are activated by the tectum after three weeks of treatment. We have found that chronic APV treatment does not alter tectal output, as indicated by the firing of isthmotectal axons, during the critical period and that chronic NMDA treatment increases tectal output in postcritical period Xenopus. Tectal output does not differ between normal Xenopus during and after the end of the critical period.(ABSTRACT TRUNCATED AT 250 WORDS)

The developing chick isthmo-optic nucleus forms a transient efferent projection to the optic tectum

The present work describes the formation of a transient efferent axonal projection from the isthmo-optic nucleus (ION) to the ipsilateral optic tectum of the chick embryo. Local application of either the carbocyanine dye DiI or rhodamine-B-isothiocyanate (RITC) to the superficial layers of the optic tectum resulted in retrograde labeling of the corresponding retinal region, and in anterograde staining of tectal axons projecting to the ION. In addition to these known projections, retrogradely labeled ION neurons appeared to be filled from the tectum. This projection, called the isthmo-tectal projection, could be characterized by means of various staining techniques: (i) It first appears at embryonic day E9 and gradually disappears after day E16. It is absent in the hatched chick. (ii) Both the cells inside the ION and those situated outside the border of the ION, the so-called ectopic cells, contribute to the formation of the isthmo-tectal projection. (iii) Double labeling from the contralateral retina (Fast blue) and from the ipsilateral tectum (DiI or RITC) revealed that some of the ION fibers projecting to the tectum are collaterals of axons normally directed to the retina. (iv) Microsurgical removal of the eye anlage early in development resulted in a numerical increase of the ION-tectal fibers. The results are discussed in terms of the role of transient projections during development.

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 retinotectal axons indirectly and that tectal cell dendrites are the critical site of interaction.

Development of topographic connections between the isthmic nuclei and optic tecta in the frog Limnodynastes dorsalis

In the frog Limnodynastes dorsalis, the pattern of topographic connections between the isthmic nuclei and optic tecta was determined by anterograde and retrograde transport of horseradish peroxidase from localised tectal regions. In both larvae and adults, reciprocal mapping of the uncrossed isthmo-tectal and tecto-isthmal projections was evidenced by the juxtaposition of labelled tecto-isthmal terminations with labelled cells in the cortex and medulla of the ipsilateral isthmic nucleus. The crossed isthmo-tectal projection was revealed by labelled cells in the cortex and medulla of the nucleus contralateral to the injection. In adults, rostral tectal areas projected to rostral and ventral regions of the ipsilateral isthmic nucleus. Following more caudal tectal injections, labelled cells were found in progressively more dorsal locations within the nucleus. Labelled cells in the contralateral nucleus were found in the rim cortex abutting a neuropil and in medullary cells adjacent to this region. Connections between ventral isthmic regions and most rostral tectum and between dorsomedial nucleus and caudomedial tectum were similar in both nuclei. However, for isthmic areas projecting to rostromedial and mid-tectum, the location of labelled cells in the contralateral nucleus was inverted with respect to the ipsilateral nucleus. This inversion would allow both nuclei to project to visually corresponding regions of each tectum. During larval stages the basic adult topography was established despite the continued neurogenesis of both isthmic nuclei and optic tecta. In late larval stages a rim neuropil appeared adjacent to the cortical region in the isthmic nuclei where labelled cells of the crossed isthmotectal projection were found. Prior to this stage labelled cells abutted labelled medullary cells. The appearance of this neuropil was approximately temporally correlated with the onset of electrophysiologically detectable responses in the ipsilateral visuotectal projection. Formation of the rim neuropil may relate to maturation of the tecto-isthmo-tectal connections which underlie this visual projection.

Plasticity in the ipsilateral visuotectal projection persists after lesions of one nucleus isthmi inXenopus

Visual input has a profound effect on the development of binocular maps in the tectum of the frog Xenopus laevis. Input from the ipsilateral eye, which is relayed to the tectum via the opposite nucleus isthmi, is normally in register with the retinotectal map from the contralateral eye. However, if one eye is rotated during larval stages while the other eye is left in normal orientation, then the resulting mismatched visual input induces the crossed isthmotectal axons to change their trajectories and to establish a reoriented ipsilateral visuotectal map in register with the contralateral retinotectal map. The major cue which aligns the two maps is the correlation of visually-evoked activity from the two eyes. This experiment was designed to determine whether the uncrossed isthmotectal projection is necessary to organize the map transmitted by the crossed isthmotectal axons. Each NI receives a topographic map from the tectum on the same side of the brain and therefore carries the same topographic information as the retinotectal projection, and each NI transmits that map not only to the opposite tectum but also back to the same tectum from which it received its input. Thus, the uncrossed isthmotectal axons provide each tectum with a map which is essentially topographically identical to the retinotecal map but which is slightly delayed temporally. The uncrossed isthmotectal axons therefore could provide topographic cues to the guide the alignment of the crossed isthmotectal axons as they establish the ipsilateral visuotectal map. In order to determine whether the uncrossed isthmotectal projection is an important source of topographic cues for the crossed isthmotectal axons, the right nucleus isthmi was ablated and one eye was rotated by 90 degrees-150 degrees in midlarval tadpoles.(ABSTRACT TRUNCATED AT 250 WORDS)

Avian nucleus isthmi ventralis projects to the contralateral optic tectum

Injections of HRP throughout the upper tectal strata led in 4 cases to the appearances of retrogradely labeled neurons within n.isthmi ventralis, contralateral to the experimental side. An additional case proved that this projection courses through the ventral supraoptic commissure. This is the first description of a crossed isthmo-tectal projection in birds.

Development of the nucleus isthmi in Xenopus, II: Branching patterns of contralaterally projecting isthmotectal axons during maturation of binocular maps

The tectum of Xenopus frogs receives input from both eyes. The contralateral eye's projection reaches the tectum directly, via the optic nerve, and the ipsilateral eye's projection reaches the tectum indirectly, via the nucleus isthmi. Under normal conditions, the topography of the ipsilateral map relayed from the nucleus isthmi is in register with the topography of the retinotectal map from the contralateral eye. During development, the process of aligning the two maps is complicated by the dramatic changes in binocular overlap of the two eyes' visual fields which take place during late tadpole and juvenile stages. The goal of this study is to determine the branching patterns of contralaterally projecting isthmotectal axons before, during, and after the period of rapid eye migration. Isthmotectal axons were filled by anterograde transport of horseradish peroxidase (HRP) from the nucleus isthmi. The results show that crossed isthmotectal axons enter the entire extent of the tectum before binocular overlap begins to increase. Therefore, binocular overlap is not necessary for the initial isthmotectal projection to span the tectum. The density of isthmotectal branches rises dramatically at the same time that the eyes begin to shift. During the period when eye migration is most rapid, many isthmotectal axons form arbors which resemble adult arbors but which extend over greater proportions of the tectal surface. The axons appear to be directed toward appropriate mediolateral positions as they enter the tectum. Their trajectories are roughly rostocaudal, with relatively little change along the mediolateral dimension. These data, when combined with available physiological data, suggest that mediolateral order is initially established by vision-independent mechanisms but can be altered by vision-dependent mechanisms. Rostrocaudal order becomes discernable only at the time when binocular visual cues become available and appears to be established primarily on the basis of the activity of the retinotectal and isthmotectal axons.

Connections between the nucleus isthmi and the tectum in larval and post-metamorphic axolotls.

The nucleus isthmi (NI) is the primary relay for the frog's ipsilateral visuotectal projection. Using electrophysiological methods, ipsilateral visuotectal activity has been recorded in thyroxine-treated, postmetamorphic axolotls but not in larval axolotls. In order to determine whether changes in isthmotectal projections are responsible for this change in electrophysiological responsiveness, we have investigated the connections between the tectum and the NI using horseradish peroxidase. Our results indicate that the axolotl's isthmotectal pathways are strikingly similar to those of the frog NI, and that the NI sends bilateral projections to the tecta in both larval and thyroxine-treated, postmetamorphic axolotls. Thus, the anatomical connections underlying the ipsilateral visuotectal projection are present during larval stages, despite the lack of electrophysiological evidence for the larval ipsilateral visuotectal projection. We hypothesize that thyroxine-induced metamorphosis produces changes in the terminal arborizations of the crossed isthmotectal projection which allow them to be detected by presynaptic electrophysiological techniques.

Visual experience and the maturation of the ipsilateral visuotectal projection in Xenopus laevis

The effect of visual deprivation upon the maturation of the ipsilateral visuotectal projection has been studied in Xenopus laevis. This topographically ordered projection is polysynaptic. The first stage involves the retinal projection to the contralateral optic tectum. The tectum projects to the nucleus isthmi on the same side. The final stage is the crossed isthmotectal projection from the nucleus isthmi to the tectum ipsilateral to the eye. The topographic precision of connections at various points in this polysynaptic pathway has been investigated by quantifying single-unit and multi-unit receptive field sizes in the contralateral and ipsilateral visuotectal projections. Observations have been made on normal animals of different ages to plot the normal maturational course of events. The effects of visual deprivation on this maturational process has been studied. Between one week and one year after metamorphosis there is an increase in the precision of connections in both the contralateral and ipsilateral visuotectal projections. Visual deprivation had no effect upon the parameters of the contralateral visuotectal projection. Ipsilateral visuotectal single units in dark-reared animals had normal receptive field sizes. Ipsilateral multi-unit receptive fields in dark-reared animals were considerably larger than in normal animals. It was concluded that the effects of visual deprivation are limited to effects on the crossed isthmotectal component of the intertectal system. In this component, however, visual experience seems to play an important role in the normal development and modification of connections. It is suggested that visual experience is utilized to accommodate changes in the system required to respond to normal changes in interocular geometry that take place with development in Xenopus.

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.

The development of the nucleus isthmi in Xenopus laevis. I. Cell genesis and the formation of connections with the tectum.

The nucleus isthmi (NI) of the amphibian relays visual input from one tectum to the other tectum and thus brings a visual map from the eye to the ipsilateral tectum. This isthmotectal visual map develops slowly; it is first detected electrophysiologically at stages 60-62, the age at which the eyes begin their dorsalward migration and the region of binocular overlap beings to increase in extent. During this critical period of life, normal binocular visual input is required for establishment of normal topographic isthmotectal projections. In this study, we have used anatomical methods to trace cell birth, cell death, and formation of connections by the nucleus isthmi during the critical period. Tritiated thymidine labelling demonstrates that cells in the nucleus isthmi are generated throughout most of tadpole life (stages 29-62). Most cells conform to an orderly ventrodorsal gradient starting from stage 29 and extending to stages 56; later cells are inserted at apparently random locations in the nucleus. We have re-examined the hypothesis of Tay and Straznicky ('80) that the order of cell genesis in the NI and tectum could help establish proper isthmotectal connections, and we find that a timing mechanisms does not explain the two-dimensional topography of the isthmotectal map but that timing may aid in proper mediolateral positioning of isthmotectal axons at the points where they first enter the tectum. Horseradish peroxidase labelling was used to investigate whether anatomical projections from tectum to NI and from NI to tectum are present prior to the onset of eye migration. The results show that there are tectoisthmotectal projections by stage 52. Moreover, isthmotectal axons grow into as yet monocular tectal regions prior to the onset of eye migration. At stage 60, when binocular overlap begins, isthmotectal axons are visible throughout the tectum but are densely branched only at the rostral tectal margin, the location where they are predicted to occur on the basis of electrophysiological maps.