Retinotectal Pathway Research Articles

In vitro reconstruction and functional development of the superior colliculus in the retinotectal pathway

In order to examine the formation of a neural network and the functional development of a visual pathway, we performed in vitro reconstruction of the retinotectal pathway using organotypic explants and co-culture methods. Retinas and superior colliculus (SC) slices obtained from embryonic rats were co-cultured on microelectrode array (MEA) substrates for four weeks. We observed retinal ganglion cell neurites innervating SC slices that evoked responses in retinas or SC slices after applying electrical stimulation. Functional connections between retinas and SC slices were formed in the cultures. At the same time, spontaneous electrical activities were recorded from both the retinas and SC slices over the four weeks. In the co-cultured SC slices, sporadic firings were initially observed at 3–4 days in vitro (DIV), and thereafter the frequency of spontaneous firing increased and synchronized activities occurred after two weeks in vitro (WIV). In most of the single-cultured SC slices, however, only sporadic firings were observed over four weeks. In addition, the retinas and SC slices were co-cultured to enable the exchange of soluble factors with each other via culture medium but not via direct neural connections. The activity patterns resembled ones of single-cultured SC slices. These results suggest that signal inputs from retinas through direct neural connections affect the development of SCs in the retinotectal pathway.

The path to learning: Action acquisition is impaired when visual reinforcement signals must first access cortex

Animals, interacting with the environment, learn and exploit the consequences of their movements. Fundamental to this is the pairing of salient sensory input with recent motor output to form an action–outcome pair linking a performed movement with its outcome. Short-latency dopamine (DA) signalling in the basal ganglia has been proposed to support this crucial task. For visual stimuli, this DA signalling is triggered at short latency by input from the superior colliculus (SC). While some aspects of the visual signal (e.g. luminance), are relayed directly to the SC via the retinotectal projection, other information unavailable to this subcortical pathway must take a more circuitous route to the SC, first submitting to early visual processing in cortex. By comparing action–outcome pairing when the visual stimulus denoting success was immediately available to the SC, via the retinotectal pathway, against that when cortical processing of the signal was required, the impact this additional sensory processing has on action–outcome learning can be established. We found that action acquisition was significantly impaired when the action was reinforced by a stimulus ineligible for the retinotectal pathway. Furthermore, we found that when the stimulus was eligible for the retinotectal pathway but evoked an increased latency, action acquisition was not impaired. These results suggest that the afferent sensory pathway via the SC is certainly primary and possibly instrumental to the DA neurons’ role in the discovery of novel actions and that the differences found are not due to simple sensory latency.

The Tumor Suppressor Gene Retinoblastoma-1 Is Required for Retinotectal Development and Visual Function in Zebrafish

Mutations in the retinoblastoma tumor suppressor gene (rb1) cause both sporadic and familial forms of childhood retinoblastoma. Despite its clinical relevance, the roles of rb1 during normal retinotectal development and function are not well understood. We have identified mutations in the zebrafish space cadet locus that lead to a premature truncation of the rb1 gene, identical to known mutations in sporadic and familial forms of retinoblastoma. In wild-type embryos, axons of early born retinal ganglion cells (RGC) pioneer the retinotectal tract to guide later born RGC axons. In rb1 deficient embryos, these early born RGCs show a delay in cell cycle exit, causing a transient deficit of differentiated RGCs. As a result, later born mutant RGC axons initially fail to exit the retina, resulting in optic nerve hypoplasia. A significant fraction of mutant RGC axons eventually exit the retina, but then frequently project to the incorrect optic tectum. Although rb1 mutants eventually establish basic retinotectal connectivity, behavioral analysis reveals that mutants exhibit deficits in distinct, visually guided behaviors. Thus, our analysis of zebrafish rb1 mutants reveals a previously unknown yet critical role for rb1 during retinotectal tract development and visual function.

Imaging Axon Pathfinding in Xenopus In Vivo

Axon pathfinding in the developing animal involves a highly dynamic process in which the axonal growth cone makes continuous decisions as it navigates toward its target. Changes occurring in the growth cone with respect to retracting from or extending into complex new territories can occur in minutes. Thus, the advent of strategies to visualize axon path-finding in vivo in a live intact animal is crucial for a better understanding of how the growth cone makes such rapid decisions in response to multiple cues. Combining these strategies with loss-of-function and/or gain-of-function techniques allows one to gain some insight as to which molecules are crucial to particular growth cone behaviors at specific choice points during navigation. The main advantage of using Xenopus lies in the accessibility of major axon tracts for live microscopy, as their embryonic development occurs ex utero. Furthermore, the robust embryos remain healthy during immobilization and allow for good imaging for long periods. This protocol describes the methods for stabilizing and preparing live Xenopus embryos for imaging labeled axonal tracts at high spatial and temporal resolution for up to 72 h. This approach can been used to investigate how the knockdown of certain gene functions can affect the speed of navigation through the well-studied Xenopus retinotectal pathway. It can be adapted to visualize other axon tracts of interest.

Visuospatial information in the retinotectal system of xenopus before correct image formation by the developing eye

The retinotectal pathway of Xenopus laevis is a well-established experimental model for studying activity-dependent processes during visual system development. Such processes can be guided by stimulus-evoked activity patterns, which depend on the refractive characteristics of the eye. Previous work has shown that many animals are hyperopic at early developmental stages due to immature refractive properties. Whether this is also the case for Xenopus laevis is unknown. Here, we measure the focal length of the lens and the size of the eye of embryos at different stages and find that Xenopus laevis exhibits a similar shift from hyperopia to emmetropia. At early stages, immediately after innervation of the tectum by the optic nerve, Xenopus embryos are hyperopic. Soon afterwards the focal length of the lens decreases and the eye converges to a state of emmetropia. Despite being hyperopic we find that some visuospatial information is available to the young circuit. Calculations based on the optical properties of the eye show that even when the animals are hyperopic the blurred retinal image provides a crude spatial resolution. Furthermore, using whole-cell recordings in the optic tectum combined with visual stimulation through the intact eye, we show that tectal neurons in hyperopic embryos have spatially restricted glutamatergic receptive fields. Our data demonstrate that Xenopus laevis eyes undergo a process of developmental emmetropization, and suggest that despite an initial stage of suboptimal image formation there is potentially enough information to guide activity-dependent refinements of the retinotectal pathway from the onset of vision.

Types of Parvalbumin-Containing Retinotectal Ganglion Cells in Mouse

The calcium-binding protein parvalbumin (PV) occurs in the retinal ganglion cells (RGCs) of various vertebrate species. In the present study, we aimed to identify the types of PV-containing RGCs that project to the superior colliculus (SC) in the mouse. We injected retrograde tracer dextran into the mouse SC to label RGCs. PV-containing RGCs were first identified by immunocytochemistry and then neurons double-labeled with dextran and PV were iontophoretically injected with a lipophilic dye, DiI. Subsequently, confocal microscopy was used to characterize the morphologic classification of the PV-immunoreactive (IR) retinotectal ganglion cells on the basis of dendritic field size, branching pattern, and stratification within the inner plexiform layer. Among the 8 different types of PV-containing RGCs in the mouse retina, we found all 8 types of RGCs projecting to the SC. The RGCs were heterogeneous in morphology. The combined approach of using tracer injection and a single cell injection after immunocytochemistry on a particular protein will provide valuable data to further understand the functional features of the RGCs which constitute the retinotectal pathway.

Intravitreous interleukin-2 treatment and inflammation modulates glial cells activation and uncrossed retinotectal development

Interleukin-2 (IL-2) plays regulatory functions both in immune and nervous system. However, in the visual system, little is known about the cellular types which respond to IL-2 and its effects. Herein, we investigated the influence of IL-2 in the development of central visual pathways. Lister Hooded rats were submitted to multiple (at postnatal days [PND]7/10/13) or single (at PND10) intravitreous injections of phosphate-buffered saline (PBS) (vehicle), zymosan, or IL-2. IL-2 receptor α subunit was detected in the whole postnatal retina. Chronic treatment with either PBS or IL-2 increases retinal glial fibrillary acidic protein (GFAP) expression, induces intravitreous inflammation revealed by the presence of macrophages, and results in a slight rearrangement of retinotectal axons. Acute zymosan treatment disrupts retinotectal axons distribution, confirming the influence of inflammation on retinotectal pathway reordering. Furthermore, acute IL-2 treatment increases GFAP expression in the retina without inflammation and produces a robust sprouting of the intact uncrossed retinotectal pathway. No difference was observed in glial cells activity in superior colliculus. Taken together, these data suggest that inflammation and interleukin-2 modulate retinal ganglion cells development and the distribution of their axons within central targets.

Identification of retinotectal projection pathway in the deep tectal laminae in the chick

The optic tectum is a visual center of nonmammalian vertebrates that receives retinal fibers in a retinotopic manner. It has been accepted that retinal fibers project to some superficial laminae of the tectum, but do not go through lamina g of stratum griseum et fibrosum superficiale (SGFS). By a novel fiber-tracing method, we found a novel pathway of retinal fibers that run through deep laminae of the tectum. The retinal fibers that would run through the newly identified pathway first run caudally along the medial edge after invading the tectum, turn laterally, and extend toward the lateral side through the deep pathway. The deep pathway runs through stratum album centrale and stratum fibrosum periventriculare. The fibers that run through the deep pathway do not enter the stratum opticum, where the conventional retinal fibers run. As development proceeds, these fibers decrease and disappear by the adult stage. By the new method, we found that some of the conventional retinal fibers transiently run through lamina g of SGFS and invade laminae h/i. In conclusion, we found distinct but transient retinal fiber pathway in the deep tectal laminae, which have not been thought to be retinorecipient.

Contribution of the retino‐tectal pathway to visually guided saccades after lesion of the primary visual cortex in monkeys

Previous reports on 'blindsight' have shown that some patients with lesions of the primary visual cortex (V1) could localize visual targets in their scotoma with hand and/or eye movements without visual awareness. A role of the retino-tectal pathway on residual vision has been proposed but the direct evidence for this still remains sparse. To examine this possibility, we inactivated the superior colliculus (SC) of unilateral V1-lesioned monkeys using microinjections of muscimol, and analysed the effects on visually guided saccades. Following muscimol injections into the contralesional SC, the monkeys performed the visually guided saccade task with relatively minor deficits. The effects of ipsilesional SC inactivation were more severe. After injections, the monkeys failed to localize the target within the visual field represented at the injection site on the SC map. The effects of ipsilesional SC inactivation may result from sensory deficits, motor deficits or a combination of both. To examine these possibilities, we tested the effects of SC inactivation on the motor system by investigating spontaneous saccades. After inactivation of the ipsilesional SC, spontaneous saccades toward the injection site were not abolished, suggesting that impairment of visually guided saccades following inactivation of the ipsilesional SC could not be explained solely by a motor deficit and was primarily due to a visual deficit, presumably by interfering with processing in the superficial layer. We conclude that the retino-tectal pathway plays an essential role in residual vision after V1 lesion. The results suggest that this pathway may be involved in mediating unconscious vision in blindsight patients.

Saccade, search and orient – the neural control of saccadic eye movements

One of the most important functions of the brain is to guide our behavior through an ever-changing environment as our goals and desires fluctuate. One simple, yet elegant system for investigating how sensory information and goal-related information combine to control and coordinate efficient behavior is the guidance of saccadic eye movements. The brain relies heavily on visual input to guide and coordinate our complex actions, but it is impossible for our brain to simultaneously process every aspect of visual information present in real world scenes at any given moment. Instead, the retina has evolved a highly specialized fovea in the center where we have the highest visual acuity. Therefore, to analyze a visual scene optimally, the eyes reorient in complex sequences so that the high acuity fovea of the retina can be directed to specific objects of interest. These saccadic eye movements are interspersed with periods of active fixation during which the visual system performs a detailed analysis of an object that may pertain to our current goals. Alternating between the serial process of making saccades and the analytical process of active fixation is repeated several hundred thousand times per day and is critical for numerous behaviors like reading this issue of EJN, driving an automobile or negotiating a busy sidewalk. In the laboratory, saccadic behavior can be measured easily and accurately. Quantitative analysis of saccade behavior can also serve as an important tool to investigate brain disorders in a host of neurological and psychiatric disorders (Leigh & Zee, 2006). Over the past few decades, there has been a tremendous increase in our knowledge of the systems that contribute to the control of saccadic eye movements. There have been many recent and exciting advances. Anatomical, physiological, clinical and imaging studies have contributed to our extensive knowledge of the saccade control circuit (Fig. 1), which includes regions of the occipital, parietal and frontal cortex, basal ganglia, thalamus, superior colliculus, cerebellum, and brainstem reticular formation. Within each of these brain regions are multiple populations of neurons and subnuclei that perform critical operations to coordinate behavior. In addition to overt eye movements that explicitly orient the visual system, visual selection may also be directed covertly to different locations or objects without any movement of the eyes. For example, we have the ability to shift attention away from where we look without initiating a saccade. Both overt (move the eyes) and covert (only shift attention) orienting can be directed voluntarily to a specific location or object, or involuntarily ‘captured’ by an abrupt change in the visual environment (Posner, 1980; Theeuwes, 1991; Fecteau & Munoz, 2006). There is significant overlap in the brain areas that participate in overt and covert orienting, which include several components of the frontoparietal network and the superior colliculus. This special issue of EJN contains reviews and original articles by some of the leading experts in the fields of saccades and visual search. There have been many recent exciting advances in these fields that are captured in the contributions in this issue. The contributions focus on different brain areas but the content ranges from the sensory input to the motor output and everything in between (Fig. 1). There are several different state-of-the-art technologies employed including quantitative behavioral analysis, neurophysiology, neuroimaging, clinical investigation, transcranial magnetic stimulation and modeling.

Lesion of Primary Visual Cortex in Monkey Impairs the Inhibitory but Not the Facilitatory Cueing Effect on Saccade

Prior visual stimulus presentation induces immediate facilitation and subsequent inhibition of orienting to an ensuing target at the same location. Recent studies revealed that the superior colliculus (SC) is involved in these facilitatory and inhibitory cueing effects on saccade; however, as the SC receives inputs both directly from the retina (retino-tectal pathway) and indirectly from visual cortices (geniculostriate pathway), it is unclear which visual pathway contributes to the effects. We investigated this issue using monkeys with lesions in the primary visual cortex (V1), thus depriving the SC of the geniculostriate pathway and leaving the retino-tectal pathway intact. We found that the inhibitory cueing effect was selectively impaired and the facilitatory cueing effect was spared after V1 lesions. The results suggest that the geniculostriate and the retino-tectal pathways are differentially involved in the generation of cueing effects on saccade: The former is critically involved in the inhibitory effect whereas the latter alone can induce the facilitatory effect. The results provide the first direct evidence for the involvement of the geniculostriate pathway in the inhibitory cueing effect and further imply that the more recent evolution of the geniculostriate pathway in higher mammals improves the efficiency of visual search by inhibiting orienting to a previously attended location.

Selective Optical Control of Synaptic Transmission in the Subcortical Visual Pathway by Activation of Viral Vector-Expressed Halorhodopsin

The superficial layer of the superior colliculus (sSC) receives visual inputs via two different pathways: from the retina and the primary visual cortex. However, the functional significance of each input for the operation of the sSC circuit remains to be identified. As a first step toward understanding the functional role of each of these inputs, we developed an optogenetic method to specifically suppress the synaptic transmission in the retino-tectal pathway. We introduced enhanced halorhodopsin (eNpHR), a yellow light-sensitive, membrane-targeting chloride pump, into mouse retinal ganglion cells (RGCs) by intravitreously injecting an adeno-associated virus serotype-2 vector carrying the CMV-eNpHR-EYFP construct. Several weeks after the injection, whole-cell recordings made from sSC neurons in slice preparations revealed that yellow laser illumination of the eNpHR-expressing retino-tectal axons, putatively synapsing onto the recorded cells, effectively inhibited EPSCs evoked by electrical stimulation of the optic nerve layer. We also showed that sSC spike activities elicited by visual stimulation were significantly reduced by laser illumination of the sSC in anesthetized mice. These results indicate that photo-activation of eNpHR expressed in RGC axons enables selective blockade of retino-tectal synaptic transmission. The method established here can most likely be applied to a variety of brain regions for studying the function of individual inputs to these regions.

Processing of low spatial frequency faces at periphery in choice reaching tasks

Various aspects of face processing have been associated with distinct ranges of spatial frequencies. Configural processing of faces depends chiefly on low spatial frequency (LSF) information whereas high spatial frequency (HSF) supports feature based processing. However, it has also been argued that face processing has a foveal-bias (HSF channels dominate the fovea). Here we used reach trajectories as a continuous behavioral measure to study perceptual processing of faces. Experimental stimuli were LSF–HSF hybrids of male and female faces superimposed and were presented peripherally and centrally. Subject reached out to touch a specified sex and their movements were recorded. The reaching trajectories reveal that there is less effect of (interference by) LSF faces at fovea as compared to periphery while reaching to HSF targets. These results demonstrate that peripherally presented LSF information, carried chiefly by magnocellular channels, enables efficient processing of faces, possibly via a retinotectal (subcortical) pathway.

Blindsight and enumeration: A case study

“Blindsight” is a term first coined by Weiskrantz et al. in 1974 to describe residual visual performance in the cortically blind. It has been postulated that blindsight could be due to the retinotectal pathway projecting information past V1 to later cortical structures. Our interest was in whether the pathways responsible for blindsight could also support enumeration. We tested a 53-year-old male, C.H., who had suffered a medial right occipital lobe stroke seven months previous. The stroke resulted in an upper left homonymous quadrantanopia. In order to determine whether there were any residual abilities in the blind field, we first tested basic detection and discrimination skills. C.H. was able to determine whether or not a 7° X 7° visual angle object was presented in his blind field with near perfect accuracy though his accuracy was only 90% for smaller (4.6° X 2.2°) figures. C.H. also discriminated between large X′s and O′s and horizontal and vertical bars with good accuracy when the figures were large (78% for X vs. O and 73% for horizontal vs. vertical bars). Throughout these tests C.H. staunchly maintained that he could not see the stimuli and that he was only guessing. Because it was clear that C.H. had some residual ability in his blind field, we tested his enumeration. The enumeration task involved 1-3 items. These items were 1.5° Blindsight and enumeration: A case study http://www.journalofvision.org/content/10/7/898.short 1 of 2 2015-01-03, 8:42 PM X 1.5° black diamonds, 0-2 in his blind field and 1-3 in his non-blind field. Across all conditions, C.H. was able to use the information in his blind field and identify the total number of items presented at levels of performance that were well above chance. Because C.H. appears to use blind field information to enumerate, we postulate that enumeration ability may be mediated by the same structures that support blindsight. Received June 26, 2010. © 2010 ARVO Articles citing this article Blliindsiight and enumeratiion:: A case study J Vis August 12, 2010 10(7): 898 Abstract Blindsight and enumeration: A case study http://www.journalofvision.org/content/10/7/898.short 2 of 2 2015-01-03, 8:42 PM

Novel retinotectal projection pathway in deeper laminae of the developing chick optic tectum

Novel retinotectal projection pathway in deeper laminae of the developing chick optic tectum

Unconscious cueing effects in saccadic eye movements – Facilitation and inhibition in temporal and nasal hemifield

The current study investigated whether subliminal spatial cues can affect the oculomotor system. In addition, we performed the experiment under monocular viewing conditions. By limiting participants to monocular viewing conditions, we can examine behavioral temporal–nasal hemifield asymmetries. These behavioral asymmetries may arise from an anatomical asymmetry in the retinotectal pathway. The results show that even though our spatial cues were not consciously perceived they did affect the oculomotor system: relative to the neutral condition, saccade latencies to the validly cued location were shorter and saccade latencies to the invalidly cued location were longer. Although we did not observe an overall inhibition of return effect, there was a reliable effect of hemifield on IOR for those observers who showed an overall IOR effect. More specifically, consistent with the notion that processing via the retinotectal pathway is stronger in the temporal hemifield than in the nasal hemifield we found an IOR effect for cues presented in the temporal hemifield but not for cues presented in the nasal hemifield. We conclude that unconsciously processed spatial cues can affect the oculomotor system. In addition, the observed behavioral temporal–nasal hemifield asymmetry is consistent with retinotectal mediation.

The role of retinotectal pathway for saccade control after the lesion of primary visual cortex

The standard approach to brain stimulation in stroke is based on the premise that ipsilesional M1 (iM1) is important for motor function of the paretic upper limb, while contralesional cortices compete with iM1. Therefore, the approach typically advocates facilitating iM1 and/or inhibiting contralesional M1 (cM1). But, this approach fails to elicit much improvement in severely affected patients, who on account of extensive damage to ipsilesional pathways, cannot rely on iM1. These patients are believed to instead rely on the undamaged cortices, especially the contralesional dorsal premotor cortex (cPMd), for support of function of the paretic limb. Here, we tested for the first time whether facilitation of cPMd could improve paretic limb function in severely affected patients, and if a cut-off could be identified to separate responders to cPMd from responders to the standard approach to stimulation.In a randomized, sham-controlled crossover study, fifteen patients received the standard approach of stimulation involving inhibition of cM1 and a new approach involving facilitation of cPMd using repetitive transcranial magnetic stimulation (rTMS). Patients also received rTMS to control areas. At baseline, impairment [Upper Extremity Fugl-Meyer (UEFMPROXIMAL, max = 36)] and damage to pathways [fractional anisotropy (FA)] was measured. We measured changes in time to perform proximal paretic limb reaching, and neurophysiology using TMS.Facilitation of cPMd generated more improvement in severely affected patients, who had experienced greater damage and impairment than a cut-off value of FA (0.5) and UEFMPROXIMAL (26–28). The standard approach instead generated more improvement in mildly affected patients. Responders to cPMd showed alleviation of interhemispheric competition imposed on iM1, while responders to the standard approach showed gains in ipsilesional excitability in association with improvement.A preliminary cut-off level of severity separated responders for standard approach vs. facilitation of cPMd.Cut-offs identified here could help select candidates for tailored stimulation in future studies so patients in all ranges of severity could potentially achieve maximum benefit in function of the paretic upper limb.

Oculomotor Distraction by Signals Invisible to the Retinotectal and Magnocellular Pathways

Irrelevant stimulus onsets interfere with saccade planning to other stimuli, prolonging saccadic latency (the oculomotor distractor effect) or eliciting directional errors (saccadic capture). Such stimulus-driven interference has been associated with the retinotectal pathway, the direct pathway from retina to superior colliculus. Consistent with this theory, the distractor effect has not been found for stimuli visible only to the short-wave cones in the retina (S cones), which are thought not to contribute to the retinotectal pathway. However, S-cone signals are generally slower than luminance signals and such differences in temporal dynamics have not been taken into account when investigating the saccadic distractor effect. Here, by varying the delay between target and distractor, we found that S-cone stimuli do in fact produce a distractor effect, but the optimal delay is generally different from that for luminance distractors. The temporal dynamics of the distractor effect conform to a general framework of saccadic competition that takes sensory transmission time into account. Additionally, we observe that S-cone stimuli are able to produce saccadic capture in our paradigm. We conclude that stimulus-driven oculomotor interference does not rely on the retinotectal pathway, or indeed the magnocellular pathway, which is also blind to our S-cone stimuli.

Purinergic modulation in the development of the rat uncrossed retinotectal pathway

Adenosine is a neuromodulator implicated in nervous system development and plasticity and its effects are mediated by inhibitory (A1, A3) and excitatory (A2a, A2b) receptors. The role of adenosine in the synaptic activity depends mainly on a balanced activation of A1 and A2a receptors which are activated by various ranges of adenosine concentrations. Herein, we investigated the expression of A1 and A2a receptors and also the accumulation of cAMP in the superior colliculus at different stages of development. Furthermore, we examined the effects of an acute in vivo blockade of adenosine deaminase during the critical period when the elimination of misplaced axons/terminals takes place with a simultaneous fine tuning of terminal arbors into appropriate terminal zones. Lister Hooded rats ranging from postnatal days (PND) 0–70 were used for ontogeny studies. Our results indicate that A1 expression in the visual layers of the superior colliculus is higher until PND 28, while A2a expression increases after PND 28 in a complementary developmental pattern. Accordingly, the incubation of collicular slices with 5′-N-ethylcarboxamido-adenosine, a non-specific adenosine receptor agonist, showed a significant reduction in cAMP accumulation at PND 14 and an increase in adults. For the anatomical studies, the uncrossed retinotectal projections were traced after the intraocular injection of horseradish peroxidase. One group received daily injections of an adenosine deaminase inhibitor (erythro-9(2-hydroxy-3-nonyl adenine), 10 mg/kg i.p.) between PND 10 and 13, while control groups were treated with vehicle injections (NaCl 0.9%, i.p.). We found that a short-term blockade of adenosine deaminase during the second postnatal week induced an expansion of retinotectal terminal fields in the rostrocaudal axis of the tectum. Taken together, the results suggest that a balance of purinergic A1 and A2a receptors through cAMP signaling plays a pivotal role during the development of topographic order in the retinotectal pathway.

Effects of different neurotrophic factors on the survival of retinal ganglion cells after a complete intraorbital nerve crush injury: A quantitative in vivo study

We examined in adult Sprague Dawley rats the loss of retinal ganglion cells (RGCs) induced by complete intraorbital optic nerve crush (IONC) as well as the effects of several neurotrophic factors to prevent IONC-induced RGC loss. Completeness of the IONC lesion was assessed by investigating the orthograde and retrograde transport of neuronal tracers applied to the origin and termination of the retinotectal pathway. RGC survival after IONC alone or combined with intraocular injection of the neurotrophic factors NT-4, BDNF or CNTF was quantified at survival intervals ranging from 5 to 12 days post-lesion (dpl) by identifying RGCs that had been pre-labelled with fluorogold (FG). RGC loss first appeared at 7 dpl and by 12 dpl only 32% of the RGC population remained in the retina. Intraocular administration of NT-4, BDNF or CNTF resulted in almost a complete protection against IONC-induced RGC loss by 7 dpl, and the protection remained significant by 12 dpl only for NT-4 and BDNF. We have analyzed these results taking into account our previous studies on the loss of RGCs induced by intraorbital optic nerve transection (IONT) and concluded that RGC loss induced by IONC is slower and less severe than that following IONT. Moreover, as for IONT-induced RGC loss, IONC-induced RGC loss may also be prevented with administration of NT-4, BDNF or CNTF, though for NT-4 and CNTF their neuroprotective effects differ depending on the injury type. Overall this data underscore the importance of the type of ON injury on the pattern of RGC degeneration as well as in their response to neuroprotective treatments.