Vision plays a crucial role in the survival of animals, and the visual system has particularly selectively evolved in response to the visual environment, ecological niche, and species habitats in vertebrate species. To date, a horizontal streak of retinal ganglion cell (RGC) distribution pattern is observed across mammal species. Here, we report that the giant panda's vertically oriented visual streak, combined with current evidence of the animal's forward-placed eyes, ocular structure, and retinal neural topographic distribution patterns, presents the emergence of a well-adapted binocular visual system. Our results suggest that the giant panda may use a unique way to processing binocular visual information. Results of mathematical simulation are in favor of this hypothesis. The topographic distribution properties of RGCs reported here could be essential for understanding the visual adaptation and evolution of this living fossil.

Purpose We aimed to observe changes in the visual function of patients with primary open-angle glaucoma (POAG) after undergoing binocular asynchronous visual training. Methods Seven patients with POAG with binocular visual field defects underwent binocular asynchronous dichoptic virtual reality (VR)-based visual perceptual training for 20 days (45 min/session/day, 5 sessions/week, for 4 weeks). Perimetry, contrast sensitivity (CS), and vision-related quality of life assessments were performed for all patients. Results Six months after completing training, nine of 14 eyes showed better performance in the perimetry (the mean deviation [MD] of perimetry was improved as compared to baseline), including six severe eyes and three mild POAG eyes. Moreover, the MD values of four of the nine eyes showed significant improvement (more than 1-dB increase as compared to baseline), including three severe eyes and one mild POAG eye. However, the MD values did not differ significantly between baseline and post-training. Contrast sensitivity tests, performed at three spatial frequencies (3, 6, and 18 cycles/degree) were significantly enhanced after asynchronous dichoptic training (p = 0.021, 0.026, and 0.020, respectively). Conclusion Patients with POAG, particularly those with severe POAG, performed significantly better in perimetry after training. All patients showed significantly improved performance on the CS tests. Improvements in visual function were sustained for at least 6 months. These results suggest that visual rehabilitation in patients with POAG can be achieved through asynchronous VR-based dichoptic visual perceptual training. A larger randomized clinical trial is required to confirm these effects. Trial registration: The trial registration number is #ChiCTR2100054625.

Amblyopia resulting from early deprivation of vision or defocus in one eye reflects an imbalance of input from the eyes to the visual cortex. We tested the hypothesis that asynchronous stimulation of the two eyes might induce synaptic plasticity and rebalance input. Experiments on normal adults showed that repetitive brief exposure of grating stimuli, with the onset of each stimulus delayed by 8.3 ms in one eye, results in a shift in perceptual eye dominance. Clinical studies(Clinical trial registration number: Chi CTR2100049130), using popular 3D movies with similar asynchrony between the two eyes(amblyopic eye stimulated first) to treat anisometropic amblyopia, established that just 10.5 h of conditioning over <3 weeks produced improvement that met criteria for successful treatment. The benefits of asynchronous conditioning accumulate over 20–30 45 min sessions, and are maintained for at least 2 years. Finally, we demonstrate that asynchronous binocular treatment alone is more effective than patching only. This novel treatment is popular with children and is some 50 times more efficient than patching alone.

Purpose To examine the effect of intraocularly applied amphiregulin antibody on physiological axial elongation in young nonhuman primates. Methods The experimental study included six male 12-months-old macaque nonhuman primates (body weight:2.46 ± 0.25kg;range:2.20-2.90kg). In the experimental group (n=3 animals), three intravitreal injections of amphiregulin antibody (100μg/50μl) were applied to the left eyes at intervals of 4-6 weeks, and injections of phosphate buffered solution (50μl) were applied to the right eyes. Three other animals were assigned to a blank control group. Results During the study period of 23.6 weeks, axial length in the experimental group did not change in the left eyes (18.91 ± 0.37mm to 18.94 ± 0.67mm; P =0.90), while it linearly increased in the right eyes (18.87 ± 0.38mm to 19.24 ± 0.53mm; P =0.056) and in the control group (left eyes:19.15 ± 0.22mm to 19.48 ± 0.22mm; P =0.009; right eyes:19.17 ± 0.15 mm to 19.46 ± 0.23 mm; P =0.024). The interocular difference in axial elongation increased in the experimental group from -0.11 ± 0.12mm at 4 weeks after baseline to -0.34 ± 0.15mm at the study end, while in the control group, the interocular side difference did not change significantly (from 0.01 ± 0.10 mm to 0.03 ± 0.08 mm; P =0.38). The difference in the interocular difference in axial elongation between the two groups was significant at 8 weeks ( P =0.01), 15 weeks ( P =0.007), and at study end ( P =0.02). The interocular difference in axial length correlated with the interocular difference in vitreous cavity length (standardized regression coefficient beta:0.85; P <0.001). The interocular axial length difference was inversely associated with the interocular refractive error difference (beta:-0.49; P <0.001). Conclusions Intraocularly applied amphiregulin antibody (100μg) reduced the physiological ocular axial elongation in juvenile nonhuman primates.

Light plays a pivotal role in the regulation of affective behaviors. However, the precise circuits that mediate the impact of light on depressive-like behaviors are not well understood. Here, we show that light influences depressive-like behaviors through a disynaptic circuit linking the retina and the lateral habenula (LHb). Specifically, M4-type melanopsin-expressing retinal ganglion cells (RGCs) innervate GABA neurons in the thalamic ventral lateral geniculate nucleus and intergeniculate leaflet (vLGN/IGL), which in turn inhibit CaMKIIα neurons in the LHb. Specific activation of vLGN/IGL-projecting RGCs, activation of LHb-projecting vLGN/IGL neurons, or inhibition of postsynaptic LHb neurons is sufficient to decrease the depressive-like behaviors evoked by long-term exposure to aversive stimuli or chronic social defeat stress. Furthermore, we demonstrate that the antidepressive effects of light therapy require activation of the retina-vLGN/IGL-LHb pathway. These results reveal a dedicated retina-vLGN/IGL-LHb circuit that regulates depressive-like behaviors and provide a potential mechanistic explanation for light treatment of depression.

The amygdala is a limbic structure that is involved in many brain functions, including emotion, learning and memory. It has been reported that melanopsin-expressing retinal ganglion cells (ipRGCs) innervate the medial amygdala (MeA). However, whether conventional RGCs (cRGCs) project to the MeA remains unknown. The goal of this study was to determine if cRGCs project to the MeA and to determine the morphological properties of MeA-projecting RGCs (MeA-RGCs). Retrogradely labeled RGCs in whole-mount retinas were intracellularly injected to reveal their dendritic morphologies. Immunohistochemical staining was performed to selectively label ipRGCs (MeA-ipRGCs) and cRGCs (MeA-cRGCs). The results showed that 95.7% of the retrogradely labeled cells were cRGCs and that the rest were ipRGCs. Specifically, MeA-cRGCs consist of two morphological types. The majority of them exhibit small but dense dendritic fields and diffuse ramification patterns as previously reported in RGB2 (95%), while the rest exhibit small but sparse dendritic branching patterns resembling those of RGB3 cells (5%). MeA-ipRGCs consist of M1 and M2 subtypes. The MeA-RGCs showed an even retinal distribution patterns. The soma and dendritic field sizes of the MeA-RGCs did not vary with eccentricity. In conclusion, the present results suggest that MeA-RGCs are structurally heterogeneous. These direct RGCs that input to the MeA could be important for regulating amygdala functions.

This study investigated visual response properties of retinal ganglion cells (RGCs) under high glucose levels. Extracellular single-unit responses of RGCs from mouse retinas were recorded. And the eyecup was prepared as a flat mount in a recording chamber and superfused with Ames medium. The averaged RF size of the ON RGCs (34.1±2.9, n=14) was significantly smaller than the OFF RGCs under the HG (49.3±0.3, n=12) (P<0.0001) conditions. The same reduction pattern was also observed in the osmotic control group (HM) between ON and OFF RGCs (P<0.0001). The averaged luminance threshold (LT) of ON RGCs increased significantly under HG or HM (HG: P<0.0001; HM: P<0.0002). OFF RGCs exhibited a similar response pattern under the same conditions (HG: P<0.01; HM: P<0.0002). The averaged contrast gain of ON cells was significantly lower than that of OFF cells with the HM treatment (P<0.015, unpaired Student’s t test). The averaged contrast gain of ON cells was significantly higher than OFF cells with the HG treatment (P<0.0001). The present results suggest that HG reduced receptive field center size, suppressed luminance threshold, and attenuated contrast gain of RGCs. The impact of HG on ON and OFF RGCs may be mediated via different mechanisms.

To characterize recombinant AAV2 (rAAV2)-mediated expression of L132C/T159C ChR2 mutant in retinal ganglion cells (RGCs) of young adult cynomolgus monkeys. rAAV2 vectors carrying a fusion construct of the ChR2 mutant and GFP (ChR2-GFP) were delivered to the vitreous chamber by intravitreal injection. Expression patterns of the ChR2 mutant in RGCs were examined by immunohistochemical methods three months after injection. The RNA-binding protein with multiple splicing (RBPMS) was used as an RGC specific marker to differentiate RGCs from other retinal neurons and non-neuronal cells. The numbers of RBPMS+ and GFP+ double-labeled RGCs in the central foveal varied with the eccentricity. The expression peaked within 100 μm from the edge of the foveola and drastically decreased to a single superficial RGC layer approximately 300 μm from the edge. On average, the ratio of the double-labeled RGCs versus RBPMS+ RGCs approached 0.32±0.15 (n=14 fields) at the central foveal region (0.1 to 0.53 mm). We observed that the ratio reached 0.78±0.16 (n=21 fields) at peripheral retinal locations (eccentricity >7 mm). This investigation demonstrates that RBPMS could serve as a valuable RGC specific marker for future investigations in this field.

PURPOSE:: Characterize the vertical distribution pattern of retinal ganglion cells in the giant panda. Methods: Four eyeballs were immediately collected from deceased adult pandas. Routine immunohistochemical staining techniques were applied to label retinal ganglion cells (RGCs). RNA-binding protein with multiple splicing (RBPMS) was used as an RGC marker to selectively label RGCs (custom-made by ProSci Inc, CA) and SMI-32 was used as an Alpha RGC marker. Immunostained RGCs were scanned under confocal microscopy and analyzed with commercial software (Adobe Photoshop and Microsoft Excel). RESULTS: As expected, the isodensity map of RGCs in the cat exhibited a typical horizontally positioned visual streak, in contrast, that of the panda showed a vertically oriented isodensity RGCs band. This band crossed the center of AC-like peak density region and extended vertically from the superior to the inferior retina (5.22 ± 0.46 mm, n=3). The averaged width of the band was 2.18 ± 0.32 mm (n=3). SMI-32 positive RGCs were also analyzed, accordingly. CONCLUSION: The present study suggests that the giant panda adopted a retinal structure that is essential for better visual performance and survival. These morphological characteristics of RGCs could be important for understanding visual adaptation and evolution of this living fossil.

Animals promote their survival by avoiding rapidly approaching objects that indicate threats. In mice, looming-evoked defensive responses are triggered by the superior colliculus (SC) which receives direct retinal inputs. However, the specific neural circuits that begin in the retina and mediate this important behaviour remain unclear. Here we identify a subset of retinal ganglion cells (RGCs) that controls mouse looming-evoked defensive responses through axonal collaterals to the dorsal raphe nucleus (DRN) and SC. Looming signals transmitted by DRN-projecting RGCs activate DRN GABAergic neurons that in turn inhibit serotoninergic neurons. Moreover, activation of DRN serotoninergic neurons reduces looming-evoked defensive behaviours. Thus, a dedicated population of RGCs signals rapidly approaching visual threats and their input to the DRN controls a serotonergic self-gating mechanism that regulates innate defensive responses. Our study provides new insights into how the DRN and SC work in concert to extract and translate visual threats into defensive behavioural responses.

PURPOSE:This study was conducted to determine whether alpha lipoic acid (ALA) promotes the survival of retinal ganglion cells (RGCs) in a rat model of optic nerve crush (ONC) injury and to investigate the neuroprotective mechanisms of ALA in the retina in this ONC injury model. METHODS:Adult male Sprague-Dawley rats (180-220 g) were subjected to ONC injury surgery. ALA (63 mg/kg) was injected intravenously 1 day before or after the ONC injury. Animals were euthanized after 10 days, and the number of ganglion cells positive for RNA-binding protein with multiple splicing (Rbpms), which is an RGC marker, were counted on the whole mount retinas. In addition, immunofluorescence and immunoblotting were performed to examine the localization and levels of erythropoietin receptor (EPOR) and neurotrophin-4/5 (NT4/5) in the retinas in all experimental groups. To determine whether the EPO/EPOR signaling pathway was involved in the ALA antioxidant pathway, the rats were subjected to ruxolitinib (INCB018424, 0.25 mg/kg, bid, intraperitoneal, i.p.) treatment after the animals were injected intravenously with ALA 1 day before ONC injury. RESULTS:The average number of Rbpms-positive cells/mm2 in the control group (sham-operated group), the ONC group, the ALA-ONC group, and the ONC-ALA group retinas was 2219±28, 418±8, 848±22, and 613±18/mm2, respectively. The ALA-ONC and ONC-ALA groups showed a statistically significantly increased RGC survival rate compared to the ONC group. There were statistical differences in the RGC survival rates between the ALA-ONC (39%) and ONC-ALA groups (28%; p<0.05). Immunofluorescent labeling showed that EPOR and NT4/5 expression was significant in the retinal ganglion cell layer (GCL). At the same time, western blot analysis revealed that ALA induced upregulation of EPOR protein and NT4/5 protein expression in the retina after ONC injury. However, INCB018424 reversed the protective effects of ALA on the ONC retinas. CONCLUSIONS:ALA has neuroprotective effects on RGCs after ONC injury. Moreover, prophylactic administration of ALA may have a stronger neuroprotective effect against ONC-induced damage. Based on these data, we also conclude that the endogenous EPO/EPOR signaling pathway may contribute to the protective effects of ALA in the retina after ONC injury.

In this study we first sought to determine whether RNA-binding protein with multiple splicing (RBPMS) can serve as a specific marker for cat retina ganglion cells (RGCs) using retrograde labeling and immunohistochemistry staining. RBPM was then used as an RGC marker to study RGC survival after optic nerve crush (ONC) and alpha-lipoic acid (ALA) treatment in cats. ALA treatment yielded a peak density of RBPMS-alpha cells within the peak isodensity zone (>60/mm2) which did not differ from ONC retinas. The area within the zone was significantly enlarged (control: 2.3%, ONC: 0.06%, ONC+ALA: 0.1%). As for the 10-21/mm2 zone, ALA treatment resulted in a significant increase in area (control: 34.5%, ONC: 12.1%, ONC+ALA: 35.9%). ALA can alleviate crush-induced RGC injury.

The dorsal raphe nucleus (DRN), the major source of serotonergic input to the forebrain, receives excitatory input from the retina that can modulate serotonin levels and depressive-like behavior. In the Mongolian gerbil, retinal ganglion cells (RGCs) with alpha-like morphological and Y-like physiological properties innervate the DRN with ON DRN-projecting RGCs out numbering OFF DRN-projecting RGCs. The DRN neurons targeted by ON and OFF RGCs are unknown. To explore retino-raphe anatomical organization, retinal afferents labeled with Cholera toxin B were examined for association with the postsynaptic protein PSD-95. Synaptic associations between retinal afferents and DRN serotonergic and GABAergic neurons were observed. To explore retino-raphe functional organization, light-evoked c-fos expression was examined. Light significantly increased the number of DRN serotonergic and GABAergic cells expressing c-Fos. When ON RGCs were rendered silent while enhancing the firing rate of OFF RGCs, c-Fos expression was greatly increased in DRN serotonergic neurons suggesting that OFF DRN-projecting RGCs predominately activate serotonergic neurons whereas ON DRN-projecting RGCs mainly target GABAergic neurons. Direct glutamatergic retinal input to DRN 5-HT neurons contributes to the complex excitatory drive regulating these cells. Light, via the retinoraphe pathway can modify DRN 5-HT neuron activity which may play a role in modulating affective behavior.

Since significant ocular differences in both anatomical structure and optical properties exist between rodents and humans, clinical imaging devices for human use are not suitable for use on rodents. In this study, we develop a contact probe with a flexible surface that can closely fit the rodent cornea for fundus imaging with a confocal scanning laser ophthalmoscope. Both Zemax simulation and in vivo fundus imaging demonstrate that this contact probe can significantly improve both the imaging quality and the operational convenience.

PURPOSE. A retinal projection into the dorsal raphe nucleus (DRN), namely, the retino-raphe projection, exists in many species. The rat is one of several species in which a retino-raphe projection has been described; however, the retinal ganglion cell (RGC) types that contribute to this pathway are unknown.METHODS. Retrograde tracing via cholera toxin subunit B (CTB) was used to reveal DRN-projecting RGCs in rats, combined with intracellular injection in vitro, melanopsin immunostaining in whole-mounted retinas, and serotonin immunostaining to define the DRN. We modified methods of CTB injection into DRN used previously in order to avoid possible contamination with other retinorecipient regions, particularly the superior colliculus (SC).RESULTS. The majority of DRN-projecting RGCs showed alpha-like morphology, and some CTB-positive RGCs were colabeled with melanopsin. Approximately 80% of the total population of CTB-labeled DRN-projecting RGCs was alpha-like cells including ON alpha cells and OFF alpha cells; these alpha-like cells were melanopsin immunonegative. Approximately 10% of the remaining DRN-projecting RGCs were melanopsin immunopositive, in which the M1 subtype of intrinsically photosensitive retinal ganglion cell (ipRGC) provided the dominant projection of ipRGCs into DRN, with only few non-M1 ipRGCs involved. The DRN-projecting ipRGCs could be retrogradely labeled following tracer injection into all rostrocaudal aspects of the DRN.CONCLUSIONS. Both conventional RGCs with alpha-like morphology and melanopsin-expressing ipRGCs project into the rat DRN. Approximately 10% of DRN-projecting RGCs were colabeled with melanopsin, and the majority of these were the M1 subtype of ipRGCs. An ipRGC component of the retino-raphe projection may contribute to a sustained light-mediated modulation of DRN serotonin release.