The role of visual experience in the formation of binocular projections in frogs

Many parts of the visual system contain topographic maps of the visual field. In such structures, the binocular portion of the visual field is generally represented by overlapping, matching projections relayed from the two eyes. One of the developmental factors which helps to bring the maps from the two eyes into register is visual input. The role of visual input is especially dramatic in the frog, Xenopus laevis. In tadpoles of this species, the eyes initially face laterally and have essentially no binocular overlap. At metamorphosis, the eyes begin to move rostrodorsally; eventually, their visual fields have a 170 degree region of binocular overlap. Despite this major change in binocular overlap, the maps from the ipsilateral and contralateral eyes to the optic tectum normally remain in register throughout development. This coordination of the two projections is disrupted by visual deprivation. In dark-reared Xenopus, the contralateral projection is nearly normal but the ipsilateral map is highly disorganized. The impact of visual input on the ipsilateral map also is shown by the effect of early rotation of one eye. Examination of the tectal lobe contralateral to the rotated eye reveals that both the contralateral and the ipsilateral maps to that tectum are rotated, even though the ipsilateral map originates from the normal eye. Thus, the ipsilateral map has changed orientation to remain in register with the contralateral map. Similarly, the two maps on the other tectal lobe are in register; in this case, both projections are normally oriented even though the ipsilateral map is from the rotated eye. The discovery that the ipsilateral eye's map reaches the tectum indirectly, via a relay in the nucleus isthmi, has made it possible to study the anatomical changes underlying visually dependent plasticity. Retrograde and anterograde tracing with horseradish peroxidase have shown that eye rotation causes isthmotectal axons to follow abnormal trajectories. An axon's route first goes toward the tectal site where it normally would arborize but then changes direction to reach a new tectal site. Such rearrangements bring the isthmotectal axons into proximity with retinotectal axons which have the same receptive fields. Anterograde horseradish peroxidase filling has also been used to study the trajectories and arborizations of developing isthmotectal axons. The results show that the axons enter the tectum before the onset of eye migration but do not begin to branch profusely until eye movement begins to create a zone of binocular space.(ABSTRACT TRUNCATED AT 400 WORDS)     

Acceleration by NMDA treatment of visually induced map reorganization in juvenile Xenopus after larval eye rotation

Each tectal lobe of Xenopus forgs receives two topographic maps, one via the ipsilateral eye and one via the contralateral eye. The alignment of the ipsilateral map with the contralateral map depends upon bincoular visual input during a critical period that extends from late tadpole to early juvenile stages. Rotation of one eye during the critical period leads to reorganization of the ipsilateral map, which eventually comes back into alignment with the contralateral map despite the abnormal eye position. The ipsilateral eye's map initially develops as if there had been no alteration in eye position; there is a delay of 4–6 weeks before reorganization can be detected by electrophysiological mapping. In this paper, the possible role of the NMDA receptor in the delay in reorganization is addressed. The degree of NMDA receptor activation may need to be above some threshold level to trigger reorganization. If NMDA receptor activation normally is below that level until after the first month postmetamorphosis, then exogenous NMDA might boost the process sufficiently to start the reorganization process sooner than usual. In order to test this possibility, the left eye of tadpoles was rotated and NMDA was applied to the right tectal lobe for 3–5 weeks, starting at 1 week postmetamorphosis. Electrophysiological mapping demonstrated that reorganization takes place more rapidly than in untreated forgs or frogs treated with vehicle only. This result is consistent with the interpretation that the activation of the NMDA receptor is a rate-limiting step in the activity-dependent matching of binocular maps in Xenopus tectum. 1994 John Wiley & Sons, Inc.     

Topographic targeting and pathfinding errors of retinal axons following overexpression of ephrinA ligands on retinal ganglion cell axons

In the retinotectal projection, the Eph receptor tyrosine kinase ligands ephrinA2 and ephrinA5 are differentially expressed not only in the tectum, but also in a high-nasal-to-low-temporal pattern in the retina. Recently, we have shown that retrovirally driven overexpression of ephrinA2 on retinal axons leads to topographic targeting errors of temporal axons in that they overshoot their normal termination zones in the rostral tectum and project onto the mid- and caudal tectum. The behavior of nasal axons, however, was only marginally affected. Here, we show that overexpression of ephrinA5 affects the topographic targeting behavior of both temporal and nasal axons. These data reinforce the idea that differential ligand expression on retinal axons contributes to topographic targeting in the retinotectal projection. Additionally, we found that ectopic expression of ephrinA2 and ephrinA5 frequently leads to pathfinding errors at the chiasm, resulting in an increased stable ipsilateral projection.     

Mutations affecting retinotectal axonal pathfinding in Medaka, Oryzias latipes

We screened for mutations affecting retinotectal axonal projection in Medaka, Oryzias latipes. In wild-type Medaka embryos, all the axons of retinal ganglion cells (RGCs) project to the contralateral tectum, such that the topological relationship of the retinal field is maintained. We labeled RGC axons using DiI/DiO at the nasodorsal and temporoventral positions of the retina, and screened for mutations affecting the pattern of stereotypic projections to the tectum. By screening 184 mutagenized haploid genomes, seven mutations in five genes causing defects in axonal pathfinding were identified, whereas mutations affecting the topographic projection of RGC axons were not found. The mutants were grouped into two classes according to their phenotypes. In mutants of Class I, a subpopulation of the RGC axons branched out either immediately after leaving the eye or after reaching the midline, and this axonal subpopulation projected to the ipsilateral tectum. In mutants of Class II, subpopulations of RGC axons branched out after crossing the midline and projected aberrantly. These mutants will provide clues to understanding the functions of genes essential for axonal pathfinding, which may be conserved or partly divergent among vertebrates.     

Binocular depth perception mechanisms in tongue-projecting salamanders

Tongue-projecting salamanders (Bolitoglossini) combine extreme speed and high precision in prey capture. They possess all requirements for stereoscopic depth perception: frontally oriented eyes, a substantial amount of direct ipsilateral projection in addition to the contralateral one, and binocularly driven neurons. Extracellular recordings were made from retinal afferents in the tectum as well as from the somata of tectal neurons. RF-sizes of afferents and tectal neurons were determined, and the response properties of tectal neurons were tested under monocular and binocular conditions with stimuli of different size and velocity. While RF-sizes and response properties of binocular neurons during binocular and contralateral stimulation were similar, ipsilaterally stimulated neurons exhibited much smaller RFs, lower spike rates and different size preferences.Furthermore, the contralateral retinotectal projection from one eye and the ipsilateral from the other are in register. While retinal afferents are distributed linearly over the tectal surface, most tectal neurons are activated by a retinal area corresponding to the frontal visual field; this results in a magnification of this region. The two monocular receptive fields of binocular neurons exhibit zero disparities (horopter) at distances that coincide with the maximum reach of the tongue. We hypothesize that bolitoglossine salamanders (as well as amphibians in general) make use of two kinds of disparities: (1) between the maps in the left and right tectal hemisphere, coding for the lateral eccentricity of an object, and (2) between the ipsilateral and contralateral retinotectal map, coding for the distance. The presence of substantial direct ipsilateral afferents in bolitoglossine salamanders appears to be the basis for a fast computation of object distance, which is characteristic of these animals.Abbreviations Ax/Ay coordinates of a recorded afference - Nx/Ny coordinates of a recorded neuron - RF receptive field - RFc contralateral receptive field - RFi ipsilateral receptive field - RFx/RFy coordinates of a receptive field center - RGC retinal ganglion cell     

Pax genes in development and maturation of the vertebrate visual system: implications for optic nerve regeneration

Pax genes play a pivotal role in development of the vertebrate visual system. Pax6 is the master control gene for eye development: ectopic expression of Pax6 in Xenopus laevis and Drosphila melanogaster leads to the formation of differentiated eyes on the legs or wings. Pax6 is involved in formation of ganglion cells of the retina, as well as cells of the lens, iris and cornea. In addition Pax6 may play a role in axon guidance in the visual system. Pax2 regulates differentiation of the optic disk through which retinal ganglion cell axons exit the eye. Furthermore, Pax2 plays a critical role in development of the optic chiasm and in the guidance of axons along the contralateral or ipsilateral tracts of the optic nerve to visual targets in the brain. During development Pax7 is expressed in neuronal cells of one of the major visual targets in the brain, the optic tectum/superior colliculus. Neurons expressing Pax7 migrate towards the pia and concentrate in the stratum griseum superficiale (SGFS), the target site for retinal axons. Together, expression of Pax2, 6 and 7 may guide axons during formation of functional retinotectal/collicular projections. Highly regulated Pax gene expression is also observed in mature animals. Moreover, evidence suggests that Pax genes are important for regeneration of the visual system. We are currently investigating Pax gene expression in species that display a range of outcomes of optic nerve regeneration. We predict that such information will provide valuable insights for the induction of successful regeneration of the optic nerve and of other regions of the central nervous system in mammals including man.     

Expression patterns of Ephs and ephrins throughout retinotectal development in Xenopus laevis

The Eph family of receptor tyrosine kinases and their ligands the ephrins play an essential role in the targeting of retinal ganglion cell axons to topographically correct locations in the optic tectum during visual system development. The African claw-toed frog Xenopus laevis is a popular animal model for the study of retinotectal development because of its amenability to live imaging and electrophysiology. Its visual system undergoes protracted growth continuing beyond metamorphosis, yet little is known about ephrin and Eph expression patterns beyond stage 39 when retinal axons first arrive in the tectum. We used alkaline phosphatase fusion proteins of EphA3, ephrin-A5, EphB2, and ephrin-B1 as affinity probes to reveal the expression patterns of ephrin-As, EphAs, ephrin-Bs, and EphBs, respectively. Analysis of brains from stage 40 to adult frog revealed that ephrins and Eph receptors are expressed throughout development. As observed in other species, staining for ephrin-As displayed a high caudal to low rostral expression pattern across the tectum, roughly complementary to the expression of EphAs. In contrast with the prevailing model, EphBs were found to be expressed in the tectum in a high dorsal to low ventral gradient in young animals. In animals with induced binocular tectal innervation, ocular dominance bands of alternating input from the two eyes formed in the tectum; however, ephrin-A and EphA expression patterns were unmodulated and similar to those in normal frogs, confirming that the segregation of axons into eye-specific stripes is not the consequence of a respecification of molecular guidance cues in the tectum.     

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.     

Convergence of multisensory inputs in Xenopus tadpole tectum

The integration of multisensory information takes place in the optic tectum where visual and auditory/mechanosensory inputs converge and regulate motor outputs. The circuits that integrate multisensory information are poorly understood. In an effort to identify the basic components of a multisensory integrative circuit, we determined the projections of the mechanosensory input from the periphery to the optic tectum and compared their distribution to the retinotectal inputs in Xenopus laevis tadpoles using dye‐labeling methods. The peripheral ganglia of the lateral line system project to the ipsilateral hindbrain and the axons representing mechanosensory inputs along the anterior/posterior body axis are mapped along the ventrodorsal axis in the axon tract in the dorsal column of the hindbrain. Hindbrain neurons project axons to the contralateral optic tectum. The neurons from anterior and posterior hindbrain regions project axons to the dorsal and ventral tectum, respectively. While the retinotectal axons project to a superficial lamina in the tectal neuropil, the hindbrain axons project to a deep neuropil layer. Calcium imaging showed that multimodal inputs converge on tectal neurons. The layer‐specific projections of the hindbrain and retinal axons suggest a functional segregation of sensory inputs to proximal and distal tectal cell dendrites, respectively. © 2009 Wiley Periodicals, Inc. Develop Neurobiol, 2009     

The Caenorhabditis elegans Six/sine oculis class homeobox gene ceh-32 is required for head morphogenesis

Repulsion plays a fundamental role in the establishment of a topographic map of the chick retinotectal projections. This has been highlighted by studies demonstrating the role of opposing gradients of the EphA3 receptor tyrosine kinase on retinal axons and two of its ligands, ephrin-A2 and ephrin-A5, in the tectum. We have analyzed the distribution of these two ephrins in other retinorecipient structures in the chick diencephalon and mesencephalon during the period when visual connections are being established. We have found that both ephrin-A2 and ephrin-A5 and their receptors EphA4 and EphA7 are expressed in gradients whose orientation is consistent with the topography of the nasotemporal axis of the respective retinofugal projections. In addition, their distribution suggests that receptor-ligand interactions may be involved in the organization of connections between the different primary visual centers and, thus, in the topographic organization of secondary visual projections. Interestingly, where projections lack a clear topographic representation, a uniform expression of the Eph-ephrin molecules was observed. Finally, we also show that a similar patterning mechanism may be implicated in the transfer of visual information to the telencephalon. These results suggest a conserved function for EphA receptors and their ligands in the elaboration of topographic maps at multiple levels of the visual pathway.     

NMDA receptor-dependent ocular dominance plasticity in adult visual cortex

The binocular region of mouse visual cortex is strongly dominated by inputs from the contralateral eye. Here we show in adult mice that depriving the dominant contralateral eye of vision leads to a persistent, NMDA receptor-dependent enhancement of the weak ipsilateral-eye inputs. These data provide in vivo evidence for metaplasticity as a mechanism for binocular competition and demonstrate that an ocular dominance shift can occur solely by the mechanisms of response enhancement. They also show that adult mouse visual cortex has a far greater potential for experience-dependent plasticity than previously appreciated. These insights may force a revision in how data on ocular dominance plasticity in mutant mice have been interpreted.     

Retinotectal plasticity in Xenopus: Anomalous ipsilateral projection following late larval eye removal

The left eye was removed from Stage 56 Xenopus tadpoles. Two to 9 months after metamorphosis, electrophysiologic analysis showed that the surviving (right) eye mediated a normal visual field projection to the left (contralateral) optic tectum. In addition, a peripheral region of the same retina innervated the entire right (ipsilateral) tectum. Primary evidence that indicates this anomalous ipsilateral projection was due to direct retina-to-tectum innervation comes from singleunit analysis, latency measurements, and tectal lesion studies. Thus, the peripheral retina simultaneously connected in much different patterns to the two optic tecta, solely on the basis of the presence (in the left tectum) or absence (in the right tectum) of central retinal fibers. This documents a role for fiber-fiber interaction (such as repulsion or competition) acting in combination with fiber-tectum interactions in the formation of the retinotectal map.     

Acute blockade of nitric oxide synthesis induces disorganization and amplifies lesion-induced plasticity in the rat retinotectal projection

In the rat visual system, the uncrossed retinotectal projection undergoes a topographical refinement within the first two postnatal weeks. We have studied the role of nitric oxide (NO), a retrograde messenger which couples pre- and postsynaptic activation, in the development of the uncrossed retinotectal projection and in the plasticity of this pathway as a result of a restricted retinal lesion in the opposite eye. During development, maximal nitric oxide synthase (NOS) activity was observed in homogenates of tectal tissue at postnatal day 5 (PND 5), followed by a two-step decrease at the end of the topographical fine tuning period (PND 21) and the adult stage (PND 42). We also tested the effects of an acute in vivo blockade of NOS during the development of both animals that had not been operated on, and lesioned animals. Animals ranging from PND 4 to PND 42 were treated either with the NOS inhibitor, L-nitro-arginine (Narg 50 mg/kg ip.) or vehicle (NaCl 0.9%) during 4 days (from PND 4-7 or PND 9-12) or 8 days (from PND 20-27 or PND 34-41). Reduction of NOS activity induced sprouting of the ipsilateral pathway up to the second postnatal week in the animals that had not been operated on. Rats that had been operated on, however, showed an amplification of the lesion-induced plasticity up to the fourth postnatal week under NOS blockade. The data suggest that NO plays a role in the stabilization of retinotectal synapses during the critical period of topographic refinement, and indicate that an acute blockade of retrograde signals enables plastic rearrangements in the visual system within this time window.     

Modulation of EphA receptor function by coexpressed ephrinA ligands on retinal ganglion cell axons

The Eph family is thought to exert its function through the complementary expression of receptors and ligands. Here, we show that EphA receptors colocalize on retinal ganglion cell (RGC) axons with EphA ligands, which are expressed in a high-nasal-to-low-temporal pattern. In the stripe assay, only temporal axons are normally sensitive for repellent axon guidance cues of the caudal tectum. However, overexpression of ephrinA ligands on temporal axons abolishes this sensitivity, whereas treatment with PI-PLC both removes ephrinA ligands from retinal axons and induces a striped outgrowth of formerly insensitive nasal axons. In vivo, retinal overexpression of ephrinA2 leads to topographic targeting errors of temporal axons. These data suggest that differential ligand expression on retinal axons is a major determinant of topographic targeting in the retinotectal projection.     

Mouse visual cortex

Neurons in mouse visual cortex have diverse receptive field properties and they respond selectively to specific features of visual stimuli. Owing to the lateral position of the eyes, only about a third of the visual cortex receives input from both eyes, but many cells in this region are binocular. Similar to higher mammals, closing one eye during a critical period shifts the responses of cells, such that they are better driven by the non-deprived eye. In this review I illustrate how the combination of transgenic mouse technology with single cell recording and modern imaging techniques might lead to a further understanding of the mechanisms that underlie the development, plasticity, and function of the mammalian visual cortex.     

Dynamic aspects of retinotectal map formation revealed by a vital-dye fiber-tracing technique

In the visual system of Xenopus laevis, the axons from the retinal ganglion cells of the eye form a topographic projection onto the optic tectum. Many studies have focused on revealing the mechanisms responsible for this precise and regular projection pattern. In contrast to the static view of the system that one might expect from examining the regularity of the projection, recent work on its regeneration and its changes during larval development indicate that part of the patterning process involves the dynamic behavior of optic fibers. Typically, anatomical and electrophysiological techniques have been used to obtain static views of the developing retinotectal projection which then must be complied to provide a glimpse of any dynamic behavior. Here we report on experiments using a newly developed fiber tracing technique to directly follow the emergence of topography in the developing retinotectal projection. Defined halves of the developing eyebud were labeled with a vital fluorescent dye which fills the growing axons, and the projection of the labeled cells was followed for up to 2 weeks in individual animals. The experiments confirm that dorsal and ventral optic nerve fibers sort out into an ordered projection early in development. In contrast, nasal and temporal fibers initially overlap, and the same sets of prelabeled fibers then sort out into the adult topography over a period of days.     

Photoreceptors and visual interneurons in the medicinal leech

The medicinal leech has five pairs of eyes, each with about 50 photoreceptors. Receptors produce propagating impulses which constitute their output to second order neurons in the CNS. Within the eye, receptors have diverse thresholds, and thus the aggregate output of the eye is graded with light intensity. By having many receptors in parallel, the eye may achieve better intensity discrimination and temporal response than would be predicted from the relatively poor characteristics of individual receptors. Receptors in eyes 3-5 on one side of the animal excite the ipsilateral LV (lateral visual) cell, an interneuron in the first segmental ganglion. By physiological tests the receptor axons are electrically coupled to the LV cell. Moreover, the LV cell is Lucifer Yellow dye-coupled to many fine fibers that appear to be receptor axons of the ipsilateral eyes 3-5. The receptors of the contralateral eyes 3-5, and those of the photosensitive sensilla lining the body inhibit the LV cell via polysynaptic pathways. Thus, the LV cells are central elements of the neural circuit processing input from the leech's spatially distributed visual system.     

Acute blockade of nitric oxide synthesis induces disorganization and amplifies lesion‐induced plasticity in the rat retinotectal projection

In the rat visual system, the uncrossed retinotectal projection undergoes a topographical refinement within the first two postnatal weeks. We have studied the role of nitric oxide (NO), a retrograde messenger which couples pre‐ and postsynaptic activation, in the development of the uncrossed retinotectal projection and in the plasticity of this pathway as a result of a restricted retinal lesion in the opposite eye. During development, maximal nitric oxide synthase (NOS) activity was observed in homogenates of tectal tissue at postnatal day 5 (PND 5), followed by a two‐step decrease at the end of the topographical fine tuning period (PND 21) and the adult stage (PND 42). We also tested the effects of an acute in vivo blockade of NOS during the development of both animals that had not been operated on, and lesioned animals. Animals ranging from PND 4 to PND 42 were treated either with the NOS inhibitor, L‐nitro‐arginine (Narg 50 mg/kg ip.) or vehicle (NaCl 0.9%) during 4 days (from PND 4–7 or PND 9–12) or 8 days (from PND 20–27 or PND 34–41). Reduction of NOS activity induced sprouting of the ipsilateral pathway up to the second postnatal week in the animals that had not been operated on. Rats that had been operated on, however, showed an amplification of the lesion‐induced plasticity up to the fourth postnatal week under NOS blockade. The data suggest that NO plays a role in the stabilization of retinotectal synapses during the critical period of topographic refinement, and indicate that an acute blockade of retrograde signals enables plastic rearrangements in the visual system within this time window. © 2000 John Wiley & Sons, Inc. J Neurobiol 44: 371–381, 2000     

Dendritic spine dynamics are regulated by monocular deprivation and extracellular matrix degradation

The mammalian primary visual cortex (V1) is especially susceptible to changes in visual input over a well-defined critical period, during which closing one eye leads to a loss of responsiveness of neurons to the deprived eye and a shift in response toward the open eye. This functional plasticity can occur rapidly, following even a single day of eye closure, although the structural bases of these changes are unknown. Here, we show that rapid structural changes at the level of dendritic spines occur following brief monocular deprivation. These changes are evident in the supra- and infragranular layers of the binocular zone and can be mimicked by degradation of the extracellular matrix with the tPA/plasmin proteolytic cascade. Further, monocular deprivation occludes a subsequent effect of matrix degradation, suggesting that this mechanism is active in vivo to permit structural remodeling during ocular dominance plasticity.