In vivo retinal imaging in translational regenerative research.

Regenerative translational studies must include a longitudinal assessment of the changes in retinal structure and function that occur as part of the natural history of the disease and those that result from the studied intervention. Traditionally, retinal structural changes have been evaluated by histological analysis which necessitates sacrificing the animals. In this review, we describe key imaging approaches such as fundus imaging, optical coherence tomography (OCT), OCT-angiography, adaptive optics (AO), and confocal scanning laser ophthalmoscopy (cSLO) that enable noninvasive, non-contact, and fast in vivo imaging of the posterior segment. These imaging technologies substantially reduce the number of animals needed and enable progression analysis and longitudinal follow-up in individual animals for accurate assessment of disease natural history, effects of interventions and acute changes. We also describe the benefits and limitations of each technology, as well as outline possible future directions that can be taken in translational retinal imaging studies.

Efficacy and safety of injecting increasing volumes into the extravascular spaces of the choroid using a blunt adjustable depth injector.

PURPOSE: To evaluate the efficacy and safety of injecting increasing volumes into the extravascular spaces of the choroid (EVSC) in rabbit eyes in vivo using a blunt adjustable depth injector. METHODS: Indocyanine green (ICG) was injected in the superior-temporal quadrant, 2 mm posterior to the limbus at increasing volumes (0.1-0.3 ml) into the EVSC of New Zealand rabbit eyes in vivo. Intraocular pressure (IOP) measurements, spectral domain optical coherence tomography (SD-OCT), fundus imaging and histology analysis were performed to assess the safety and efficacy of the injection. RESULTS: Volumes up to 0.3 ml were administered consistently. ICG injection was successfully monitored in vivo using infrared fundus imaging and SD-OCT. ICG was detected across the EVSC compartment, reaching the retinal pigment epithelium, optic nerve head and visual streak. Injection of 0.3 ml yielded maximal dye distribution with a coverage area of 61.8% ± 6.7% (mean ± standard error, SE) of the posterior segment. Maximal IOP elevation was recorded 5 min following injection of 0.2 and 0.3 ml ICG (+ 20.0 mmHg, + 19.4 mmHg, respectively). Twenty minutes post-injection, the IOP was < 15 mmHg in all injection volumes. No retinal detachment or hemorrhages were detected in any of the injected eyes. CONCLUSIONS: This study demonstrates consistent and safe delivery of large volumes within the EVSC using a blunt adjustable depth injector that distributes the dye over 60% of the retinal surface. This injection system may offer a minimally invasive and easy way to deliver large volumes of pharmaceuticals into the posterior segment.

Vision in chameleons-A model for non-mammalian vertebrates.

Chameleons (Chamaeleonidae, Reptilia) are known for their extreme sensory and motor adaptations to arboreal life and insectivoury. They show most distinct sequences of visuo-motor patterns in threat avoidance and in predation with prey capture being performed by tongue strikes that are unparalleled in vertebrates. Optical adaptations result in retinal image enlargement and the unique capacity to determine target distance by accommodation cues. Ocular adaptations result in complex eye movements that are context dependent, not independent, as observed in threat avoidance and predation. In predation, evidence from the chameleons' capacity to track multiple targets support the view that their eyes are under individual controls. Eye movements and body movements are lateralised, with lateralisation being a function of many factors at the population, individual, and specific-situation levels. Chameleons are considered a potentially important model for vision in non-mammalian vertebrates. They provide exceptional behavioural tools for studying eye movements as well as information gathering and analysis. They open the field of lateralisation, decision making, and context dependence. Finally, chameleons allow a deeper examination of the relationships between their unique visuo-motor capacities and the central nervous system of reptiles and ectotherms, in general, as compared with mammals.

In vivo MRI assessment of bioactive magnetic iron oxide/human serum albumin nanoparticle delivery into the posterior segment of the eye in a rat model of retinal degeneration.

BACKGROUND: Retinal degeneration diseases affect millions of patients worldwide and lead to incurable vision loss. These diseases are caused by pathologies in the retina and underlying choroid, located in the back of the eye. One of the major challenges in the development of treatments for these blinding diseases is the safe and efficient delivery of therapeutics into the back of the eye. Previous studies demonstrated that narrow size distribution core-shell near infra-red fluorescent iron oxide (IO) nanoparticles (NPs) coated with human serum albumin (HSA, IO/HSA NPs) increase the half-life of conjugated therapeutic factors, suggesting they may be used for sustained release of therapeutics. In the present study, the in vivo tracking by MRI and the long term safety of IO/HSA NPs delivery into the suprachoroid of a rat model of retinal degeneration were assessed. RESULTS: Twenty-five Royal College of Surgeons (RCS) pigmented rats received suprachoroidal injection of 20-nm IO/HSA NPs into the right eye. The left eye was not injected and used as control. Animals were examined by magnetic resonance imaging (MRI), electroretinogram (ERG) and histology up to 30 weeks following injection. IO/HSA NPs were detected in the back part of the rats' eyes up to 30 weeks following injection by MRI, and up to 6 weeks by histology. No significant differences in retinal structure and function were observed between injected and non-injected eyes. There was no significant difference in the weight of IO/HSA NP-injected animals compared to non-injected rats. CONCLUSIONS: MRI could track the nanoparticles in the posterior segment of the injected eyes demonstrating their long-term persistence, and highlighting the possible use of MRI for translational studies in animals and in future clinical studies. Suprachoroidal injection of IO/HSA NPs showed no sign of adverse effects on retinal structure and function in a rat model of retinal degeneration, suggesting that suprachoroidal delivery of IO/HSA NPs is safe and that these NPs may be used in future translational and clinical studies for extended release drug delivery at the back of the eye.

Avoidance of a moving threat in the common chameleon (Chamaeleo chamaeleon): rapid tracking by body motion and eye use.

A chameleon (Chamaeleo chamaeleon) on a perch responds to a nearby threat by moving to the side of the perch opposite the threat, while bilaterally compressing its abdomen, thus minimizing its exposure to the threat. If the threat moves, the chameleon pivots around the perch to maintain its hidden position. How precise is the body rotation and what are the patterns of eye movement during avoidance? Just-hatched chameleons, placed on a vertical perch, on the side roughly opposite to a visual threat, adjusted their position to precisely opposite the threat. If the threat were moved on a horizontal arc at angular velocities of up to 85°/s, the chameleons co-rotated smoothly so that (1) the angle of the sagittal plane of the head relative to the threat and (2) the direction of monocular gaze, were positively and significantly correlated with threat angular position. Eye movements were role-dependent: the eye toward which the threat moved maintained a stable gaze on it, while the contralateral eye scanned the surroundings. This is the first description, to our knowledge, of such a response in a non-flying terrestrial vertebrate, and it is discussed in terms of possible underlying control systems.

Lateralization of visually guided detour behaviour in the common chameleon, Chamaeleo chameleon, a reptile with highly independent eye movements.

Chameleons (Chamaeleonidae, reptilia), in common with most ectotherms, show full optic nerve decussation and sparse inter-hemispheric commissures. Chameleons are unique in their capacity for highly independent, large-amplitude eye movements. We address the question: Do common chameleons, Chamaeleo chameleon, during detour, show patterns of lateralization of motion and of eye use that differ from those shown by other ectotherms? To reach a target (prey) in passing an obstacle in a Y-maze, chameleons were required to make a left or a right detour. We analyzed the direction of detours and eye use and found that: (i) individuals differed in their preferred detour direction, (ii) eye use was lateralized at the group level, with significantly longer durations of viewing the target with the right eye, compared with the left eye, (iii) during left side, but not during right side, detours the durations of viewing the target with the right eye were significantly longer than the durations with the left eye. Thus, despite the uniqueness of chameleons' visual system, they display patterns of lateralization of motion and of eye use, typical of other ectotherms. These findings are discussed in relation to hemispheric functions.

Relating lateralization of eye use to body motion in the avoidance behavior of the chameleon (Chamaeleo chameleon).

Lateralization is mostly analyzed for single traits, but seldom for two or more traits while performing a given task (e.g. object manipulation). We examined lateralization in eye use and in body motion that co-occur during avoidance behaviour of the common chameleon, Chamaeleo chameleon. A chameleon facing a moving threat smoothly repositions its body on the side of its perch distal to the threat, to minimize its visual exposure. We previously demonstrated that during the response (i) eye use and body motion were, each, lateralized at the tested group level (N = 26), (ii) in body motion, we observed two similar-sized sub-groups, one exhibiting a greater reduction in body exposure to threat approaching from the left and one--to threat approaching from the right (left- and right-biased subgroups), (iii) the left-biased sub-group exhibited weak lateralization of body exposure under binocular threat viewing and none under monocular viewing while the right-biased sub-group exhibited strong lateralization under both monocular and binocular threat viewing. In avoidance, how is eye use related to body motion at the entire group and at the sub-group levels? We demonstrate that (i) in the left-biased sub-group, eye use is not lateralized, (ii) in the right-biased sub-group, eye use is lateralized under binocular, but not monocular viewing of the threat, (iii) the dominance of the right-biased sub-group determines the lateralization of the entire group tested. We conclude that in chameleons, patterns of lateralization of visual function and body motion are inter-related at a subtle level. Presently, the patterns cannot be compared with humans' or related to the unique visual system of chameleons, with highly independent eye movements, complete optic nerve decussation and relatively few inter-hemispheric commissures. We present a model to explain the possible inter-hemispheric differences in dominance in chameleons' visual control of body motion during avoidance.

Visually guided avoidance in the chameleon (Chamaeleo chameleon): response patterns and lateralization.

The common chameleon, Chamaeleo chameleon, is an arboreal lizard with highly independent, large-amplitude eye movements. In response to a moving threat, a chameleon on a perch responds with distinct avoidance movements that are expressed in its continuous positioning on the side of the perch distal to the threat. We analyzed body-exposure patterns during threat avoidance for evidence of lateralization, that is, asymmetry at the functional/behavioral levels. Chameleons were exposed to a threat approaching horizontally from the left or right, as they held onto a vertical pole that was either wider or narrower than the width of their head, providing, respectively, monocular or binocular viewing of the threat. We found two equal-sized sub-groups, each displaying lateralization of motor responses to a given direction of stimulus approach. Such an anti-symmetrical distribution of lateralization in a population may be indicative of situations in which organisms are regularly exposed to crucial stimuli from all spatial directions. This is because a bimodal distribution of responses to threat in a natural population will reduce the spatial advantage of predators.