Kevin R. Duffy

Darkness Alters Maturation of Visual Cortex and Promotes Fast Recovery from Monocular Deprivation

The existence of heightened brain plasticity during critical periods in early postnatal life is a central tenet of developmental sensory neuroscience and helps explain the enduring deficits induced by early abnormal sensory exposure. The human visual disorder amblyopia has been linked to unbalanced visual input to the two eyes in early postnatal visual cortical development and has been modeled in animals by depriving them of patterned visual input to one eye, a procedure known as monocular deprivation (MD). We investigated the possibility that a period of darkness might reset the central visual pathways to a more plastic stage and hence increase the capacity for recovery from early MD. Here we show that a 10 day period of complete darkness reverses maturation of stable cytoskeleton components in kitten visual cortex and also results in rapid elimination of, or even immunity from, visual deficits linked to amblyogenic rearing by MD. The heightened instability of the cytoskeleton induced by darkness likely represents just one of many parallel molecular changes that promote visual recovery, possibly by release of the various brakes on cortical plasticity.

The case from animal studies for balanced binocular treatment strategies for human amblyopia

Although amblyopia typically manifests itself as a monocular condition, its origin has long been linked to unbalanced neural signals from the two eyes during early postnatal development, a view confirmed by studies conducted on animal models in the last 50 years. Despite recognition of its binocular origin, treatment of amblyopia continues to be dominated by a period of patching of the non‐amblyopic eye that necessarily hinders binocular co‐operation. This review summarizes evidence from three lines of investigation conducted on an animal model of deprivation amblyopia to support the thesis that treatment of amblyopia should instead focus upon procedures that promote and enhance binocular co‐operation. First, experiments with mixed daily visual experience in which episodes of abnormal visual input were pitted against normal binocular exposure revealed that short exposures of the latter offset much longer periods of abnormal input to allow normal development of visual acuity in both eyes. Second, experiments on the use of part‐time patching revealed that purposeful introduction of episodes of binocular vision each day could be very beneficial. Periods of binocular exposure that represented 30–50% of the daily visual exposure included with daily occlusion of the non‐amblyopic could allow recovery of normal vision in the amblyopic eye. Third, very recent experiments demonstrate that a short 10 day period of total darkness can promote very fast and complete recovery of visual acuity in the amblyopic eye of kittens and may represent an example of a class of artificial environments that have similar beneficial effects. Finally, an approach is described to allow timing of events in kitten and human visual system development to be scaled to optimize the ages for therapeutic interventions.

Recovery of neurofilament following early monocular deprivation

Postnatal development of the mammalian geniculostriate visual pathway is partly guided by visually driven activity. Disruption of normal visual input during certain critical periods can alter the structure of neurons, as well as their connections and functional properties. Within the layers of the dorsal lateral geniculate nucleus (dLGN), a brief early period of monocular deprivation can alter the structure and soma size of neurons within deprived-eye-receiving layers. This modification of structure is accompanied by a marked reduction in labeling for neurofilament protein, a principle component of the stable cytoskeleton. This study examined the extent of neurofilament recovery in monocularly deprived cats that either had their deprived eye opened (binocular recovery), or had the deprivation reversed to the fellow eye (reverse occlusion). The loss of neurofilament and the reduction of soma size caused by monocular deprivation were ameliorated equally and substantially in both recovery conditions after 8 days. The degree to which this recovery was dependent on visually driven activity was examined by placing monocularly deprived animals in complete darkness. Though monocularly deprived animals placed in darkness showed recovery of soma size in deprived layers, the manipulation catalyzed a loss of neurofilament labeling that extended to non-deprived layers as well. Overall, these results indicate that both recovery of soma size and neurofilament labeling is achieved by removal of the competitive disadvantage of the deprived eye. However, while the former occurred even in the absence of visually driven activity, recovery of neurofilament did not. The finding that a period of darkness produced an overall loss of neurofilament throughout the dLGN suggests that this experiential manipulation may cause the visual pathways to revert to an earlier more plastic developmental stage. It is possible that short periods of darkness could be incorporated as a component of therapeutic measures for treatment of deprivation-induced disorders such as amblyopia.

Monocular deprivation provokes alteration of the neuronal cytoskeleton in developing cat lateral geniculate nucleus

Monocular deprivation early in development produces considerable change in the organization of connections within the central mammalian visual system. In the dorsal lateral geniculate nucleus, the soma, dendrites, and axon terminal fields of deprived cells become considerably smaller than nondeprived counterparts. We have examined the possibility that subcellular events enabling structural modification of deprived neurons include modification of proteins comprising the cytoskeleton. We examined the integrity of the cytoskeleton by measuring the response of a subset of its proteins to varying durations of monocular deprivation. Loss of all three neurofilament subunits (light, medium, and heavy) within deprived layers was observed to parallel changes in neuron gross structure. Monocular deprivation initiated beyond early life produced neither a change in structure nor a loss of neurofilament labeling.

Receptive field properties of neurons in the primary visual cortex under photopic and scotopic lighting conditions

Knowledge of the physiology of the primate visual cortex (area V-1) comes mostly from studies done in photopic conditions, in which retinal cones are active and rods play little or no part. Conflicting results have come from research into the effects of dark adaptation on receptive field organization of cells in the retina and the lateral geniculate nucleus. These studies claim either that the effect of the surround disappears with dark adaptation or that it does not. The current study has as its objective a comparison of responses of V-1 cells in awake-alert macaque monkeys under conditions of light and dark adaptation. We reasoned that basic receptive field properties of V-1 cells such as orientation selectivity, direction selectivity, and end-stopping should be preserved in scotopic conditions if the receptive field organization of antecedent cells is maintained in dim light. Our results indicate that dark adaptation does not alter basic V-1 receptive field characteristics such as selectivity for orientation, direction, and bar length.

Recovery of visual functions in amblyopic animals following brief exposure to total darkness.

Occlusion of one eye of kittens (monocular deprivation) results in a severe and permanent loss of visual acuity in that eye, which parallels closely the vision loss characteristic of human amblyopia. We extended earlier work to demonstrate that amblyopic vision loss can be either blocked or erased very fast by a 10 day period of total darkness following a period of monocular deprivation that begins near birth and extends to at least 8 weeks of age. The parameters of darkness were strict because no visual recovery was observed after 5 days of darkness. In addition, short periods of light introduced each day during an otherwise 10 day period of darkness obliterated the benefits. Despite recovery of normal visual acuity, only one‐quarter of the animals showed evidence of having attained normal stereoscopic vision. A period of total darkness may catalyse and improve treatment outcomes in amblyopic children.

Experience-dependent changes in NMDAR1 expression in the visual cortex of an animal model for amblyopia.

When normal binocular visual experience is disrupted during postnatal development, it affects the maturation of cortical circuits and often results in the development of poor visual acuity known as amblyopia. Two main factors contribute to the development of amblyopia: visual deprivation and reduced binocular competition. We investigated the affect of these two amblyogenic factors on the expression of the NMDAR1 subunit in the visual cortex because activation of the NMDA receptor is a key mechanism of developmental neural plasticity. We found that disruption of binocular correlations by monocular deprivation promoted a topographic loss of NMDAR1 expression within the cortical representations of the central visual field and the vertical and horizontal meridians. In contrast, binocular deprivation, which primarily affects visual deprivation, promoted an increase in NMDAR1 expression throughout the visual cortex. These different changes in NMDAR1 expression can be described as topographic and homeostatic plasticity of NMDA expression, respectively. In addition, the changes in NMDA expression in the visual cortex provide a greater understanding of the neural mechanisms that underlie the development of amblyopia and the potential for visual recovery.

Cytoskeleton alteration correlates with gross structural plasticity in the cat lateral geniculate nucleus

Monocular deprivation during early development causes rearrangement of neural connections within the visual cortex that produces a shift in ocular dominance favoring the non-deprived eye. This alteration is manifested anatomically within deprived layers of the lateral geniculate nucleus (LGN) where neurons have smaller somata and reduced geniculocortical terminal fields compared to non-deprived counterparts. Experiments using monocular deprivation have demonstrated a spatial correlation between cytoskeleton alteration and morphological change within the cat LGN, raising the possibility that subcellular events mediating deprivation-related structural rearrangement include modification to the neuronal cytoskeleton. In the current study we compared the spatial and temporal relationships between cytoskeleton alteration and morphological change in the cat LGN. Cross-sectional soma area and neurofilament labeling were examined in the LGN of kittens monocularly deprived at the peak of the critical period for durations that ranged from 1 day to 7 months. After 4 days of deprivation, neuron somata within deprived layers of the LGN were significantly smaller than those within non-deprived layers. This structural change was accompanied by a spatially coincident reduction in neurofilament immunopositive neurons that was likewise significant after 4 days of deprivation. Both anatomical effects reached close to their maximum by 10 days of deprivation. Results from this study demonstrate that alteration to the neuronal cytoskeleton is both spatially and temporally linked to the gross structural changes induced by monocular deprivation.

Binocular eyelid closure promotes anatomical but not behavioral recovery from monocular deprivation

Deprivation of patterned vision of frontal eyed mammals early in postnatal life alters structural and functional attributes of neurones in the central visual pathways, and can produce severe impairments of the vision of the deprived eye that resemble the visual loss observed in human amblyopia. A traditional approach to treatment of amblyopia has been the occlusion of the stronger fellow eye in order to force use of the weaker eye and thereby strengthen its connections in the visual cortex. Although this monocular treatment strategy can be effective at promoting recovery of visual acuity of the amblyopic eye, such binocular visual functions as stereoscopic vision often remain impaired due in part to the lack of concordant vision during the period of unilateral occlusion. The recent development of binocular approaches for treatment of amblyopia that improve the possibility for binocular interaction have achieved success in promoting visual recovery. The full and rapid recovery of visual acuity observed in amblyopic kittens placed in complete darkness is an example of a binocular treatment whose success may in part derive from a restored balance of visually-driven neural activity. In the current study we examined as an alternative to dark rearing the efficacy of binocular lid suture (BLS) to stimulate anatomical and visual recovery from a preceding amblyogenic period of monocular deprivation. In the dorsal lateral geniculate nucleus (dLGN) of monocularly deprived kittens, darkness or BLS for 10days produced a complete recovery of neurone soma size within initially deprived layers. The growth of neurone somata within initially deprived dLGN layers after darkness or BLS was accompanied by an increase in neurotrophin-4/5 labeling within these layers. Although anatomical recovery was observed in both recovery conditions, BLS failed to promote any improvement of the visual acuity of the deprived eye no matter whether it followed immediately or was delayed with respect to the prior period of monocular deprivation. Notwithstanding the lack of visual recovery with BLS, all animals in the BLS condition that were subsequently placed in darkness exhibited a substantial recovery of visual acuity in the amblyopic eye. We conclude that the balanced binocular visual input provided by BLS does not stimulate the collection of neural events necessary to support recovery from amblyopia. The complete absence of visually-driven activity that occurs with dark rearing evidently plays an important role in the recovery process.

Rapid recovery from the effects of early monocular deprivation is enabled by temporary inactivation of the retinas

Significance Normal development of the visual cortex depends critically on early life visual experience. In humans, a disparity in the quality of vision between two eyes during infancy or early childhood leads to a visual impairment called amblyopia. In animal models, amblyopia can be induced early in life using a brief period of monocular deprivation via eyelid closure. Here we show in two evolutionarily divergent species that experimental amblyopia can be rapidly corrected if binocular visual experience is restored after temporarily silencing the retinas with a local anesthetic. These findings point the way to an approach for clinical management of amblyopia with advantages over the current standard of care. A half-century of research on the consequences of monocular deprivation (MD) in animals has revealed a great deal about the pathophysiology of amblyopia. MD initiates synaptic changes in the visual cortex that reduce acuity and binocular vision by causing neurons to lose responsiveness to the deprived eye. However, much less is known about how deprivation-induced synaptic modifications can be reversed to restore normal visual function. One theoretically motivated hypothesis is that a period of inactivity can reduce the threshold for synaptic potentiation such that subsequent visual experience promotes synaptic strengthening and increased responsiveness in the visual cortex. Here we have reduced this idea to practice in two species. In young mice, we show that the otherwise stable loss of cortical responsiveness caused by MD is reversed when binocular visual experience follows temporary anesthetic inactivation of the retinas. In 3-mo-old kittens, we show that a severe impairment of visual acuity is also fully reversed by binocular experience following treatment and, further, that prolonged retinal inactivation alone can erase anatomical consequences of MD. We conclude that temporary retinal inactivation represents a highly efficacious means to promote recovery of function.