Retinocollicular Map Research Articles

Roles of Ephrin-As and Structured Activity in the Development of Functional Maps in the Superior Colliculus

The orderly projections from retina to superior colliculus (SC) preserve a continuous retinotopic representation of the visual world. The development of retinocollicular maps depend on a combination of molecular guidance cues and patterned neural activity. Here, we characterize the functional retinocollicular maps in mice lacking the guidance molecules ephrin-A2, -A3, and -A5 and in mice deficient in both ephrin-As and structured spontaneous retinal activity, using a method of Fourier imaging of intrinsic signals. We find that the SC of ephrin-A2/A3/A5 triple knock-out mice contains functional maps that are disrupted selectively along the nasotemporal (azimuth) axis of the visual space. These maps are discontinuous, with patches of SC responding to topographically incorrect locations. The patches disappear in mice that are deficient in both ephrin-As and structured activity, resulting in a near-absence of azimuth map in the SC. These results indicate that ephrin-As guide the formation of functional topography in the SC, and patterned retinal activity clusters cells based on their correlated firing patterns. Comparison of the SC and visual cortical mapping defects in these mice suggests that although ephrin-As are required for mapping in both SC and visual cortex, ephrin-A-independent mapping mechanisms are more important in visual cortex than in the SC.

Implication of neuropilin 2/semaphorin 3F in retinocollicular map formation

Neural representations of the environment within the brain take the form of topographic maps whose formation relies on graded expression of axon guidance molecules. Retinocollicular map formation, from retinal ganglion cells (RGCs) to the superior colliculus (SC) in the midbrain, is mainly driven by Eph receptors and their ligands ephrins. However, other guidance molecules participate in the formation of this map. Here we demonstrate that the receptor Neuropilin-2 is expressed in an increasing nasal-temporal gradient in RGCs, whereas one of its ligands, Semaphorin3F, but not other Sema3 molecules, presents a graded low-rostral to high-caudal expression in the SC when mapping is underway. Neuropilin-2 and its coreceptor Plexin A1 are present on RGC growth cones. Collapse assays demonstrate that Semaphorin3F induces significant growth cone collapse of temporal, but not nasal, RGCs expressing high levels of Neuropilin-2. Our results suggest that Neuropilin-2/Semaphorin3F are new candidates involved in retinotopy formation within the SC.

Mechanisms of response homeostasis during retinocollicular map formation

The mechanisms of Hebbian synaptic plasticity have been widely hypothesized to play a role in the activity-dependent development of neural circuits. However, these mechanisms are inherently unstable and would lead to the runaway excitation or depression of circuits if left unchecked. In the last decade, a number of elegant studies have demonstrated that homeostatic plasticity mechanisms exist to stabilize neural networks and maintain the constancy of neuronal output in response to changes in activity levels. These include synaptic scaling, sliding threshold models of synaptic plasticity, dynamic regulation of the number and strength of synapses, and bidirectional control of intrinsic excitability. Recently, we showed that the total synaptic input onto individual neurons of the mouse superior colliculus is preserved regardless of the size of their visual receptive fields, a phenomenon we term 'response homeostasis'. Here, we argue that regulating the capacity for synaptic plasticity and controlling the number and strength of retinocollicular inputs can preserve collicular neuron output, and we present evidence that changes in intrinsic excitability are not associated with response homeostasis. We also review findings from a number of different mutant mice and discuss whether and how different cellular mechanisms may underlie response homeostasis. Combined with other studies, our work reveals an important role for homeostatic mechanisms in regulating functional connectivity during the construction of receptive fields and the refinement of topographic maps.

Bone Morphogenetic Proteins, Eye Patterning, and Retinocollicular Map Formation in the Mouse

Patterning events during early eye formation determine retinal cell fate and can dictate the behavior of retinal ganglion cell (RGC) axons as they navigate toward central brain targets. The temporally and spatially regulated expression of bone morphogenetic proteins (BMPs) and their receptors in the retina are thought to play a key role in this process, initiating gene expression cascades that distinguish different regions of the retina, particularly along the dorsoventral axis. Here, we examine the role of BMP and a potential downstream effector, EphB, in retinotopic map formation in the lateral geniculate nucleus (LGN) and superior colliculus (SC). RGC axon behaviors during retinotopic map formation in wild-type mice are compared with those in several strains of mice with engineered defects of BMP and EphB signaling. Normal RGC axon sorting produces axon order in the optic tract that reflects the dorsoventral position of the parent RGCs in the eye. A dramatic consequence of disrupting BMP signaling is a missorting of RGC axons as they exit the optic chiasm. This sorting is not dependent on EphB. When BMP signaling in the developing eye is genetically modified, RGC order in the optic tract and targeting in the LGN and SC are correspondingly disrupted. These experiments show that BMP signaling regulates dorsoventral RGC cell fate, RGC axon behavior in the ascending optic tract, and retinotopic map formation in the LGN and SC through mechanisms that are in part distinct from EphB signaling in the LGN and SC.

Developmental homeostasis of mouse retinocollicular synapses.

Spontaneous retinal waves during development are thought to provide an instructive signal for precise retinotopic mapping by correlating the activity of neighboring retinal ganglion cells. In mutant mice (beta2-/-) that lack correlated waves, retinocollicular map refinement is impaired. In vivo recordings reveal that neurons in the superior colliculus of beta2-/- mice have large receptive fields and low peak visual responses, resulting in a conservation of total integrated response. We find that this "response homeostasis" is maintained on a cell-by-cell basis, and argue that it does not depend on regulation from the visual cortex during adulthood. Instead, in vitro recordings show that homeostasis arises from the conservation of total synaptic input from the retina, and that it is maintained via different mechanisms over development. In the absence of correlated retinal waves, beta2-/- neurons sample a larger number of weaker retinal inputs relative to controls after the first postnatal week. Once retinal waves are restored, developmental learning rules and homeostasis drive refinement so that fewer, stronger synapses are retained, as in wild-type mice, but from a larger retinal area. Homeostasis in neurons has been shown previously to regulate the gain of synaptic transmission in response to perturbations of activity. Our results suggest that during the development of sensory maps, a unique consequence of homeostatic mechanisms is the precise shaping of neuronal receptive fields in conjunction with activity-dependent competition.

Ephrin-As and Patterned Retinal Activity Act Together in the Development of Topographic Maps in the Primary Visual System

The development of topographic maps in the primary visual system is thought to rely on a combination of EphA/ephrin-A interactions and patterned neural activity. Here, we characterize the retinogeniculate and retinocollicular maps of mice mutant for ephrins-A2, -A3, and -A5 (the three ephrin-As expressed in the mouse visual system), mice mutant for the beta2 subunit of the nicotinic acetylcholine receptor (that lack early patterned retinal activity), and mice mutant for both ephrin-As and beta2. We also provide the first comprehensive anatomical description of the topographic connections between the retina and the dorsal lateral geniculate nucleus. We find that, although ephrin-A2/A3/A5 triple knock-out mice have severe mapping defects in both projections, they do not completely lack topography. Mice lacking beta2-dependent retinal activity have nearly normal topography but fail to refine axonal arbors. Mice mutant for both ephrin-As and beta2 have synergistic mapping defects that result in a near absence of map in the retinocollicular projection; however, the retinogeniculate projection is not as severely disrupted as the retinocollicular projection is in these mutants. These results show that ephrin-As and patterned retinal activity act together to establish topographic maps, and demonstrate that midbrain and forebrain connections have a differential requirement for ephrin-As and patterned retinal activity in topographic map development.

Pax7 and superior collicular polarity: insights from Pax6 (Sey) mutant mice

Pax genes are important modulators of CNS development. Pax7 and Pax6 polarise the neural tube and regionalise the brain. Pax7 is pivotal in specifying the superior colliculus/tectum, an important centre for integration of visuomotor responses and a target for Pax6+ retinal ganglion cell axons during retinocollicular mapping. Whilst initial Pax7-specification of the mesencephalon is well-established, a role in regulating polarity within the maturing mouse superior colliculus is yet to be defined, although already detailed for the chick tectum. We therefore quantified Pax7 cellular distribution and expression levels at three functionally distinct stages of superior collicular development, and analysed Pax7 expression in response to aberrant axonal input and altered forebrain/midbrain boundary placement in Pax6 mutant mice. Comparative expression profiles of ephrin-A2 and its co-localisation with Pax7 were determined in wildtype and Pax6 mutant mice. Results indicate that graded Pax7 expression in wildtype mice is perturbed in Pax6 mutant mice; changes manifest as a shift in polarity, loss of graded expression and dramatically reduced protein levels during RGC synaptogenesis. Ephrin-A2 expression is similarly altered. These results implicate Pax7 as an important determinant of polarity within the mouse superior colliculus, and suggest a role in retinotopic mapping.

Analysis of mouse EphA knockins and knockouts suggests that retinal axons programme target cells to form ordered retinotopic maps

I present a novel analysis of abnormal retinocollicular maps in mice in which the distribution of EphA receptors over the retina has been modified by knockin and/or knockout of these receptor types. My analysis shows that in all these cases, whereas the maps themselves are discontinuous, the graded distribution of EphA over the nasotemporal axis of the retina is recreated within the pattern of axonal terminations across rostrocaudal colliculus. This suggests that the guiding principle behind the formation of ordered maps of nerve connections between vertebrate retina and superior colliculus, or optic tectum, is that axons carrying similar amounts of Eph receptor terminate near to one another on the target structure. I show how the previously proposed marker induction model embodies this principle and predicts these results. I then describe a new version of the model in which the properties of the markers, or labels, are based on those of the Eph receptors and their associated ligands, the ephrins. I present new simulation results, showing the development of maps between two-dimensional structures, exploring the role of counter-gradients of labels across the target and confirming that the model reproduces the retinocollicular maps found in EphA knockin/knockout mice. I predict that abnormal distributions of label within the retina lead to abnormal distributions of label over the target, so that in each of the types of knockin/knockout mice analysed, there will be a different distribution of labels over the target structure. This mechanism could be responsible for the flexibility with which neurons reorganise their connections during development and the degree of precision in the final map. Activity-based mechanisms would play a role only at a later stage of development to remove the overlap between individual retinal projection fields, such as in the development of patterns of ocular dominance stripes.

Opposing Gradients of Ephrin-As and EphA7 in the Superior Colliculus Are Essential for Topographic Mapping in the Mammalian Visual System

During development of the retinocollicular projection in mouse, retinal axons initially overshoot their future termination zones (TZs) in the superior colliculus (SC). The formation of TZs is initiated by interstitial branching at topographically appropriate positions. Ephrin-As are expressed in a decreasing posterior-to-anterior gradient in the SC, and they suppress branching posterior to future TZs. Here we investigate the role of an EphA7 gradient in the SC, which has the reverse orientation to the ephrin-A gradient. We find that in EphA7 mutant mice the retinocollicular map is disrupted, with nasal and temporal axons forming additional or extended TZs, respectively. In vitro, retinal axons are repelled from growing on EphA7-containing stripes. Our data support the idea that EphA7 is involved in suppressing branching anterior to future TZs. These findings suggest that opposing ephrin-A and EphA gradients are required for the proper development of the retinocollicular projection.

Guidelines for reporting statistics in journals published by the American Physiological Society

Guidelines for reporting statistics in journals published by the American Physiological Society

A stochastic model for retinocollicular map development

BackgroundWe examine results of gain-of-function experiments on retinocollicular maps in knock-in mice [Brown et al. (2000) Cell 102:77]. In wild-type mice the temporal-nasal axis of retina is mapped to the rostral-caudal axis of superior colliculus. The established map is single-valued, which implies that each point in retina maps to a unique termination zone in superior colliculus. In homozygous Isl2/EphA3 knock-in mice the map is double-valued, which means that each point on retina maps to two termination zones in superior colliculus. This is because about 50 percent of cells in retina express Isl2, and two types of projections, wild-type and Isl2/EphA3 positive, form two branches of the map. In heterozygous Isl2/EphA3 knock-ins the map is intermediate between the homozygous and wild-type: it is single-valued in temporal and double-valued in the nasal parts of retina. In this study we address possible reasons for such a bifurcation of the map.ResultsWe study the map formation using stochastic model based on Markov chains. In our model the map undergoes a series of reconstructions with probabilities dependent upon a set of chemical cues. Our model suggests that the map in heterozygotes is single-valued in temporal region of retina for two reasons. First, the inhomogeneous gradient of endogenous receptor in retina makes the impact of exogenous receptor less significant in temporal retina. Second, the gradient of ephrin in the corresponding region of superior colliculus is smaller, which reduces the chemical signal-to-noise ratio. We predict that if gradient of ephrin is reduced by a genetic manipulation, the single-valued region of the map should extend to a larger portion of temporal retina, i.e. the point of transition between single-and doulble-valued maps should move to a more nasal position in Isl2-EphA3 heterozygotes.ConclusionsWe present a theoretical model for retinocollicular map development, which can account for intriguing behaviors observed in gain-of-function experiments by Brown et al., including bifurcation in heterozygous Isl2/EphA3 knock-ins. The model is based on known chemical labels, axonal repulsion/competition, and stochasticity. Possible mapping in Isl2/EphB knock-ins is also discussed.

Retinotopic Map Refinement Requires Spontaneous Retinal Waves during a Brief Critical Period of Development

During retinocollicular map development, spontaneous waves of action potentials spread across the retina, correlating activity among neighboring retinal ganglion cells (RGCs). To address the role of retinal waves in topographic map development, we examined wave dynamics and retinocollicular projections in mice lacking the β2 subunit of the nicotinic acetylcholine receptor. β2 −/− mice lack waves during the first postnatal week, but RGCs have high levels of uncorrelated firing. By P8, the wild-type retinocollicular projection remodels into a refined map characterized by axons of neighboring RGCs forming focal termination zones (TZs) of overlapping arbors. In contrast, in P8 β2 −/− mice, neighboring RGC axons form large TZs characterized by broadly distributed arbors. At P8, glutamatergic retinal waves appear in β2 −/− mice, and later, visually patterned activity appears, but the diffuse TZs fail to remodel. Thus, spontaneous retinal waves that correlate RGC activity are required for retinotopic map remodeling during a brief early critical period.

Molecular determinants of retinal ganglion cell development, survival, and regeneration.

The retina is an easily accessible part of the CNS with a well-defined cytological architecture. It allows for detailed study of the regulation of neurogenesis, determinants of cell fate specification, and signals for cell survival versus programmed cellular death during development. Within the retina, retinal ganglion cells (RGCs) are the only neurons connecting to the brain. Their axonal projection to the midbrain targets, the superior colliculus (SC), and the lateral geniculate nucleus (LGN) has been subject of a number of investigations, and led to the identification of molecular signals directing topographic information for precise wiring during development. Transcription factors, guidance molecules, extracellular matrix proteins, neurotrophic factors, and cell death-regulating factors of the Bcl-2 family and caspases, have all been reported to be involved in the processes of formation of a precise retino-collicular map, and regulation of developmental cell death.During adulthood, RGCs and their projection have to be maintained, since-to our current knowledge-they cannot be replaced following injury. On the other hand, insults of various kinds can be potentially hazardous to RGCs. Therefore, much work has been directed towards understanding of the molecular regulation of RGC degeneration following insults such as retinal ischaemia, axonal lesion, or in optic neuropathy. Experimental strategies are being devised towards protection of lesioned RGCs. Since following axonal lesion, these cells not only need to survive, but also have to reconnect in order to be functionally relevant, efforts are directed towards not only survival, but also axonal regeneration and proper rewiring of injured RGCs. This paper reviews the molecular determinants of RGC fate determination and the development of the retino-tectal projection. We summarize what is known (and hypothesized) on the determinants of RGC survival during normal adulthood, and the mechanisms of RGC degeneration in the injured retina. We also try to develop perspectives towards neuroprotection and regeneration of adult lesioned RGCs that may be applicable to lesioned CNS neurons in vertebrates in a broader sense.

NMDA antagonists in the superior colliculus prevent developmental plasticity but not visual transmission or map compression.

Partial ablation of the superior colliculus (SC) at birth in hamsters compresses the retinocollicular map, increasing the amount of visual field represented at each SC location. Receptive field sizes of single SC neurons are maintained, however, preserving receptive field properties in the prelesion condition. The mechanism that allows single SC neurons to restrict the number of convergent retinal inputs and thus compensate for induced brain damage is unknown. In this study, we examined the role of N-methyl-D-aspartate (NMDA) receptors in controlling retinocollicular convergence. We found that chronic 2-amino-5-phosphonovaleric acid (APV) blockade of NMDA receptors from birth in normal hamsters resulted in enlarged single-unit receptive fields in SC neurons from normal maps and further enlargement in lesioned animals with compressed maps. The effect was linearly related to lesion size. These results suggest that NMDA receptors are necessary to control afferent/target convergence in the normal SC and to compensate for excess retinal afferents in lesioned animals. Despite the alteration in receptive field size in the APV-treated animals, a complete visual map was present in both normal and lesioned hamsters. Visual responsiveness in the treated SC was normal; thus the loss of compensatory plasticity was not due to reduced visual responsiveness. Our results argue that NMDA receptors are essential for map refinement, construction of receptive fields, and compensation for damage but not overall map compression. The results are consistent with a role for the NMDA receptor as a coincidence detector with a threshold, providing visual neurons with the ability to calculate the amount of visual space represented by competing retinal inputs through the absolute amount of coincidence in their firing patterns. This mechanism of population matching is likely to be of general importance during nervous system development.

Cellular Localization of Ephrin-A2, Ephrin-A5, and Other Functional Guidance Cues Underlies Retinotopic Development across Species

Avian retinotectal and rodent retinocollicular systems are general model systems used to examine developmental processes that underpin topographically organized neuronal circuits. The two systems rely on guidance components to establish their precise retinotopic maps, but many cellular events differ during their development. For example, compared with the chick, a generally less restricted outgrowth pattern is observed when retinae innervate their targets in rodents. Cellular or molecular distributions of guidance components may account for such differences in retinotopic development across species. Candidate repellent molecules, such as ephrin-A2 and ephrin-A5, have been cloned in both chick and rodents; however, it has not yet been shown in rodents that living cells express sufficient amounts of any repellent components to deter outgrowth. We used a coculture assay that gives cellular resolution of retinotarget interactions and demonstrate that living, caudal superior colliculus cells selectively prevent extension of axons from temporal regions of the retinae. Time-lapse video microscopy revealed the cellular localization of permissive and repulsive guidance components in rodents, which differed from that in chick. To analyze the potential molecular basis for these differences, we investigated the function and localization of ephrin-A2 and -A5. Cells transfected with ephrin-A2 and -A5 selectively repelled retinal axons. Ephrin-A2 and -A5 RNA expression patterns differed across cell populations and between species, suggesting molecular mechanisms and key cellular interactions that may underlie fundamental differences in the development of retinotectal and retinocollicular maps.

Chapter 26 The effect of damage of the brachium of the superior colliculus in neonatal and adult hamsters and the use of peripheral nerve to restore retinocollicular projections

Using horseradish peroxidase (HRP) tracing technique, we were able to confirm the critical age in hamsters as reported previously (SO et al., 1981). Thus, following transection of the retinal fibers at the brachium of the superior colliculus (BSC) on postnatal-day 4 (P4) or later, no retinocollicular projections were observed in the adult stage. However, the retinal fibers were observed to reinnervate the superior colliculus (SC) if the BSC was cut on P3 or earlier. Physiological recording showed a close to normal retinocollicular map following a BSC damage on P0. Although retinal fibers did not reinnervate the SC following a BSC cut on or after P4, they could be observed to grow along a membrane over the damaged site. Bridging the site of BSC damage in adult hamsters using a segment of peripheral nerve (PN), retinal fibers labelled with WGA-HRP were observed to reinnervate the SC along the PN graft and visual evoked responses could be recorded in the SC showing the PN graft is effective in restoring damaged central visual pathways in adult mammals.

Regulation of NMDA receptor mRNA during visual map formation and after receptor blockade.

The topographic refinement of the rat retinocollicular projection is dependent on normal NMDA receptor function. Here we examined the expression of NMDA and non-NMDA glutamate receptor mRNA in the rat superior colliculus (SC) during this postnatal refinement period. The temporal expression pattern of mRNA coding for the NMDA receptor subunit NR1 in the superficial SC followed the time course of collicular synaptogenesis. A pronounced increase of NR1 mRNA levels occurred during the late stages of retinocollicular map refinement. In cortex, the time course of the expression of NR1 mRNA in cortex was found to be similar to that observed in SC, with low levels during the first postnatal week, a maximum at P19, and a decrease thereafter. In SC, but not in cortex, there was a change in the ratio of the two NR1 transcripts during the second postnatal week that parallels a previously demonstrated developmental change in the mean open time of NMDA channels in collicular neurons. In contrast, the mRNA expression pattern of the non-NMDA receptor subunit GluR2 in the developing SC was not closely correlated with synaptic changes. Chronic treatment of the SC with the NMDA receptor antagonist 2-amino-5-phosphonovalerate (APV) for 12 or 19 days, which disrupts retinocollicular map formation, appears to block the developmental rise in NR1 mRNA levels. These findings support a specific role for the NMDA receptor subtype of glutamate receptors in the control of synaptogenesis and developmental plasticity in the SC.

The locus of optic nerve head representation in the chick retinotectal map lacks a retinal projection

Retinotopic representation of the optic nerve head of the contralateral eye lies at the rostrodorsal face of the avian tectum. Since the homologous mammalian retino-collicular map shows an optic disc gap, the present anterograde HRP transport experiments were designed to detect an equivalent gap in the avian tectal retinorecipient strata. Sections tangential to the tectum at the locus of pecten representation displayed a thin, elongated, projection-less strip, crossing radially all retinorecipient laminae. Histochemical demonstration of NADH-diaphorase on similar sections showed low activity of this enzyme along an equivalent, elongated strip.