Effects of environmental enrichment on GLUT expression in the visual cortex of amblyopic rats.

OBJECTIVE: To investigate the potential changes of glucose metabolism and glucose transporter protein (GLUT) in the visual cortex of formally deprived amblyopic rats, as well as the effects of enriched environments on the levels of nerve conduction and glucose metabolism in the visual cortex of amblyopic rats. METHODS: 36 rats were randomly divided into three groups: CON + SE (n = 12), MD + SE (n = 12) and MD + EE (n = 12). The right eyelids of both MD + SE and MD + EE groups were sutured. After successful modelling, the MD + EE group was maintained in an enriched environment, and the other two groups were kept in the same environment. Pattern visual evoked potentials (PVEP) was used to confirm models' effect, glucose metabolism was analyzed by Micro-PET/CT (18F-FDG), and the protein as well as mRNA expression levels of GLUT were detected by Western Blot and quantitative RT-PCR (quantitative Reverse Transcription-Polymerase Chain Reaction) analyses, site of GLUT expression by immunofluorescence (IF). RESULTS: After suture modelling, both the MD + EE and MD + SE groups objective visual nerve conduction function decreased, the glucose metabolism in the visual cortex was markedly lower. After the enriched environment intervention, it recovered in the MD + EE group. The expression levels of GLUT1 and GLUT3 were increased in the MD + EE group in comparison with the MD + SE group. GLUT1 was primarily expressed on astrocytes and endothelial cells, but GLUT3 was mainly expressed on neurons. CONCLUSION: Enrichment of the environment exhibited a therapeutic effect on amblyopia, which could be related to the enhancement of glucose metabolism and GLUT expression in the visual cortex.

Visual experience has opposing influences on the quality of stimulus representation in adult primary visual cortex.

Transient dark exposure, typically 7-10 days in duration, followed by light reintroduction is an emerging treatment for improving the restoration of vision in amblyopic subjects whose occlusion is removed in adulthood. Dark exposure initiates homeostatic mechanisms that together with light-induced changes in cellular signaling pathways result in the re-engagement of juvenile-like plasticity in the adult such that previously deprived inputs can gain cortical territory. It is possible that dark exposure itself degrades visual responses, and this could place constraints on the optimal duration of dark exposure treatment. To determine whether eight days of dark exposure has a lasting negative impact on responses to classic grating stimuli, neural activity was recorded before and after dark exposure in awake head-fixed mice using two-photon calcium imaging. Neural discriminability, assessed using classifiers, was transiently reduced following dark exposure; a decrease in response reliability across a broad range of spatial frequencies likely contributed to the disruption. Both discriminability and reliability recovered. Fixed classifiers were used to demonstrate that stimulus representation rebounded to the original, pre-deprivation state, thus dark exposure did not appear to have a lasting negative impact on visual processing. Unexpectedly, we found that dark exposure significantly stabilized orientation preference and signal correlation. Our results reveal that natural vision exerts a disrupting influence on the stability of stimulus preference for classic grating stimuli and, at the same time, improves neural discriminability for both low and high-spatial frequency stimuli.

Effect of regulation of the NRG1/ErbB4 signaling pathway on the visual cortex synaptic plasticity of amblyopic adult rats.

This study aimed to investigate the effect of the neuregulin-1/epidermal growth factor 4 (NRG1/ErbB4) signaling pathway on visual cortex synaptic plasticity in adult amblyopic rats with monocular deprivation (MD). Compared with the control group, the P wave latency and amplitude of the MD group were prolonged and low, respectively, with reduced synaptic plasticity-related protein expression, lower number of visual cortex neurons, and increased apoptosis of visual cortex neurons. Recombinant neuregulin-1 (rNRG1) administration activated the NRG1/ErbB4 signaling pathway and improved the visual cortex synaptic plasticity in MD amblyopic rats. However, the effects of rNRG1 were reversed by AG1478 (ErbB4 receptor blockers). The NRG1/ErbB4 signaling pathway in the parvalbumin neurons from MD rats was also inactivated. Amblyopic rats had significantly low cell activity and downregulated expression of synaptic plasticity-related proteins. Thus, exogenous administration of NRG1 can activate ErbB4 signal transduction and improve the damaged synaptic plasticity of the visual cortex among amblyopic rats. Further studies are warranted to explore the potential for clinical management of amblyopia.

Synaptosomal Actin Dynamics in the Developmental Visual Cortex Regulate Behavioral Visual Acuity in Rats.

Purpose: Synaptosomal actin dynamics are essential for synaptic structural stability. Whether actin dynamics are involved in structural and functional synaptic plasticity within the primary visual cortex (V1) or behavioral visual acuity in rats has still not been thoroughly investigated. Methods: Synaptosome preparation and western blot analysis were used to analyze synaptosomal actin dynamics. Transmission electron microscopy was used to detect synaptic density and mitochondrial area alterations. A visual water maze task was applied to assess behavioral visual acuity. Microinjection of the actin polymerization inhibitor or stabilizer detected the effect of actin dynamics on visual function. Results: Actin dynamics, the mitochondrial area, and synaptic density within the area of V1 are increased during the critical period for the development of binocularity. Microinjection of the actin polymerization inhibitor cytochalasin D into the V1 decreased the mitochondrial area, synaptic density, and behavioral visual acuity. Long-term monocular deprivation reduced actin dynamics, the mitochondrial area, and synaptic density within the V1 contralateral to the deprived eye compared with those ipsilateral to the deprived eye and impaired visual acuity in the amblyopic eye. In addition, the mitochondrial area, synaptic density, and behavioral visual acuity were improved by stabilization of actin polymerization by jasplakinolide microinjection. Conclusions: During the critical period of visual development of binocularity, synaptosomal actin dynamics regulate synaptic structure and function and play roles in behavioral visual acuity in rats.

Subanesthetic Ketamine Reactivates Adult Cortical Plasticity to Restore Vision from Amblyopia.

Subanesthetic ketamine evokes rapid and long-lasting antidepressant effects in human patients. The mechanism for ketamine's effects remains elusive, but ketamine may broadly modulate brain plasticity processes. We show that single-dose ketamine reactivates adult mouse visual cortical plasticity and promotes functional recovery of visual acuity defects from amblyopia. Ketamine specifically induces downregulation of neuregulin-1 (NRG1) expression in parvalbumin-expressing (PV) inhibitory neurons in mouse visual cortex. NRG1 downregulation in PV neurons co-tracks both the fast onset and sustained decreases in synaptic inhibition to excitatory neurons, along with reduced synaptic excitation to PV neurons in vitro and in vivo following a single ketamine treatment. These effects are blocked by exogenous NRG1 as well as PV targeted receptor knockout. Thus, ketamine reactivation of adult visual cortical plasticity is mediated through rapid and sustained cortical disinhibition via downregulation of PV-specific NRG1 signaling. Our findings reveal the neural plasticity-based mechanism for ketamine-mediated functional recovery from adult amblyopia.

Homeostatic regulation of perisynaptic matrix metalloproteinase 9 (MMP9) activity in the amblyopic visual cortex.

Dark exposure (DE) followed by light reintroduction (LRx) reactivates robust synaptic plasticity in adult mouse primary visual cortex (V1), which allows subsequent recovery from amblyopia. Previously we showed that perisynaptic proteolysis by MMP9 mediates the enhancement of plasticity by LRx in binocular adult mice (Murase et al., 2017). However, it was unknown if a visual system compromised by amblyopia could engage this pathway. Here we show that LRx to adult amblyopic mice induces perisynaptic MMP2/9 activity and extracellular matrix (ECM) degradation in deprived and non-deprived V1. Indeed, LRx restricted to the amblyopic eye is sufficient to induce robust MMP2/9 activity at thalamo-cortical synapses and ECM degradation in deprived V1. Two-photon live imaging demonstrates that the history of visual experience regulates MMP2/9 activity in V1, and that DE lowers the threshold for the proteinase activation. The homeostatic reduction of the MMP2/9 activation threshold by DE enables visual input from the amblyopic pathway to trigger robust perisynaptic proteolysis.

Evaluation of Metabolite Changes in the Occipital Cortex of Patients with Idiopathic Infantile Nystagmus or Bilateral Ametropic Amblyopia by Magnetic Resonance Spectroscopy.

PURPOSE: To evaluate the effects of idiopathic infantile nystagmus (IN) and bilateral ametropic amblyopia on metabolites in the occipital cortex by magnetic resonance spectroscopy. METHODS: The children included in this prospective study were divided into three groups. Group 1 consisted of 11 patients with idiopathic IN, group 2 consisted of 10 patients with bilateral ametropic amblyopia and group 3 consisted of nine normal children. A single-voxel magnetic resonance spectroscopy examination was performed by placing a region of interest on the occipital cortex of each participant. N-acetyl aspartate (NAA), creatine (Cr) and choline (Cho) concentrations were measured in the occipital cortex. This was followed by calculating and comparing the NAA/Cr and Cho/Cr ratios between the three groups. The Kruskal-Wallis test, Mann-Whitney U-test, and chi-square test were used for statistical analysis. RESULTS: There was no statistically significant difference in NAA/Cr ratios between patients with idiopathic IN and normal children, but there was a statistically significant difference between these groups when Cho/Cr ratios were compared; the ratio was higher in the idiopathic IN group. There were no statistically significant differences in NAA/Cr or Cho/Cr ratios between patients with bilateral ametropic amblyopia and normal children. CONCLUSIONS: Our findings suggest that the neurochemical profile of the occipital cortex is partially affected by idiopathic IN, but not by bilateral ametropic amblyopia.

Classification of Visual Cortex Plasticity Phenotypes following Treatment for Amblyopia.

Monocular deprivation (MD) during the critical period (CP) has enduring effects on visual acuity and the functioning of the visual cortex (V1). This experience-dependent plasticity has become a model for studying the mechanisms, especially glutamatergic and GABAergic receptors, that regulate amblyopia. Less is known, however, about treatment-induced changes to those receptors and if those changes differentiate treatments that support the recovery of acuity versus persistent acuity deficits. Here, we use an animal model to explore the effects of 3 visual treatments started during the CP (n = 24, 10 male and 14 female): binocular vision (BV) that promotes good acuity versus reverse occlusion (RO) and binocular deprivation (BD) that causes persistent acuity deficits. We measured the recovery of a collection of glutamatergic and GABAergic receptor subunits in the V1 and modeled recovery of kinetics for NMDAR and GABAAR. There was a complex pattern of protein changes that prompted us to develop an unbiased data-driven approach for these high-dimensional data analyses to identify plasticity features and construct plasticity phenotypes. Cluster analysis of the plasticity phenotypes suggests that BV supports adaptive plasticity while RO and BD promote a maladaptive pattern. The RO plasticity phenotype appeared more similar to adults with a high expression of GluA2, and the BD phenotypes were dominated by GABAA α1, highlighting that multiple plasticity phenotypes can underlie persistent poor acuity. After 2-4 days of BV, the plasticity phenotypes resembled normals, but only one feature, the GluN2A:GluA2 balance, returned to normal levels. Perhaps, balancing Hebbian (GluN2A) and homeostatic (GluA2) mechanisms is necessary for the recovery of vision.

Retinal oximetry in normal and amblyopic children: a pilot study.

PURPOSE: To study the retinal vascular oxygen saturation in amblyopic eyes and compare them to unaffected fellow eyes and eyes of normal subjects. METHODS: A total of 32 amblyopic, 24 normal and 13 fellow eyes of patients below age of 18 were enrolled in this prospective observation study. Retinal oximetry was performed using the Oxymap T1 retinal oximeter. Retinal vascular oxygen saturations and diameters were compared between amblyopic eyes, normal eyes and unaffected fellow eyes. RESULTS: The average age was 8.6 years in the amblyopia group (M:F 16:16) and 10.9 years in the normal group (M:F 7:5; p = 0.024). Median corrected distance visual acuity in the amblyopia group was 20/50; it was 20/20 in the other groups (p < 0.001). The average arteriolar and venous saturation in the amblyopia, normal and fellow group was 84.5% (95% CI: 82.6-86.4), 83.2% (95% CI: 80.7-85.6) and 80.8% (95% CI: 78.6-82.9) and 51.9% (95% CI: 50.4-53.4), 50.8% (95% CI: 48.2-53.4) and 49.3% (95% CI: 45.7-52.9). There was no statistically significant difference between the saturation values of the amblyopia group and the controls, however, significantly higher values were found in the amblyopia group compared to the fellow group for arteriolar and venous saturations (p = 0.013; p = 0.005). Arteriolar and venous diameters showed no significant difference between groups. CONCLUSION: Amblyopic eyes showed higher mean oxygen saturations than the fellow eyes. This observation could be due to altered neuronal activity or could be due to a measurement artefact due to alterations in retinal reflectivity.

Transcranial direct-current stimulation (tDCS) improves detection of simple bright stimuli by amblyopic Long Evans rats in the SLAG task and produces an increase of parvoalbumin labelled cells in visual cortices.

In this work visual functional improvement of amblyopic Long Evans rats treated with tDCS has been assessed using the "slow angled-descent forepaw grasping" (SLAG) test. This test is based on an innate response that does not requires any memory-learning component and has been used before for measuring visual function in rodents. The results obtained show that this procedure is useful to assess monocular but not binocular deficits, as controls and amblyopic animals showed significant differences during monocular but not during binocular assessment. On the other hand, parvoalbumin labelling was analysed in three areas of the visual cortex (V1M, V1B and V2L) before and after tDCS treatment. No changes in labelling were observed after monocular deprivation. However, tDCS treatment significantly improved vision through the amblyopic eye, and a significant increase of parvoalbumin-positive cells was observed in the three areas, both in the stimulated hemisphere but also in the non-stimulated hemisphere. This effect occurred both in control and amblyopic animals. Thus, tDCS induced changes are similar in controls and amblyopic animals, although only the last one showed a functional improvement.

Amblyopia, Strabismus and Refractive Errors in Congenital Ptosis: a systematic review and meta-analysis.

Congenital ptosis may be associated with abnormalities of visual development and function, including amblyopia, strabismus and refractive errors. However, the prevalence estimates of these abnormalities vary widely. We performed a systematic review and meta-analysis to estimate the prevalence of amblyopia, strabismus and refractive errors in congenital ptosis. Cochrane, Pubmed, Medline, Embase, and Web of Science were searched by July 2017. We used random/fixed effects models based on a proportion approach to estimate the prevalence. Heterogeneity would be considered signifcant if the p values less than 0.1 and/or I2 greater than 50%. Subgroup analyses, meta-regression analyses and sensitivity analyses were utilized to explore the potential sources of it. A total of 24 studies selected from 3,633 references were included. The highest prevalence was revealed for myopia with 30.2% (95%CI 3.0-69.8%), followed by 22.7% (95%CI 18.5-27.8%) for amblyopia, 22.2% (95%CI 7.8-63.1%) for astigmatism, 19.6% (95%CI 16.5-23.2%) for strabismus, 17.3% (95% CI 13.1-22.9%) for anisometropia and 4.0% for hyperopia (95%CI 1.8-7.1%). Significant heterogeneity was identified across most estimates. Our findings suggest that amblyopia, strabismus and refractive errors in congenital ptosis are present in much higher percentage. This study highlights the importance of early diagnosis and timely treatment of patients with congenital ptosis.

Expression of nitric oxide synthase in the retina of monocular deprivation amblyopia rats.

OBJECTIVE: Amblyopia or lazy eye is a common visual problem affecting children that cannot correct with lenses. Nitric oxide synthase (NOS) is a critical enzyme that regulates the activity of nitric oxide (NO), a key signaling molecule with multiple roles in many tissues. Among its many activities, NOS has been proposed to be required for normal eye development and altered NOS expression can lead to perturbations in eye development and vision. MATERIALS AND METHODS: To examine the potential role of neuronal NOS (nNOS) in vision loss, we generated a model of monocular deprivation amblyopia in rats. After suturing one eye, we examined several parameters of neural activity and nNOS expression in the retina 7, 14 and 28 days later. RESULTS: We found the rapid and progressive loss of neural activity in the retina of sutured eyes compared to non-treated and control eyes. The sutured eyes also showed decreased expression of nNOS at the protein and mRNA levels, indicating a strong correlation between nNOS expression and retina activity. CONCLUSIONS: These data suggest a potential role for nNOS activity in vision loss, opening potential avenues for therapeutic intervention.

Efficacy of an Amblyopia Treatment Program with Both Eyes Open: A Functional Near-Infrared Spectroscopy Study.

INTRODUCTION AND PURPOSE: To investigate the efficacy of an amblyopia treatment program with both eyes open. METHODS: Ten subjects (mean age 20.5 ± 1.5 years) were enrolled. All subjects had un-remarkable ophthalmic examinations, but several subjects had minor refractive errors. Vision function was evaluated using the 3-D visual function trainer-ORTe. Brain measurements were made using functional near-infrared spectroscopy (fNIRS) to examine the oxygenated hemoglobin (HbO2) concentration change upon visual stimulus presentation. The three conditions were as follows: both eyes open and both eyes stimulated, both eyes open and only one eye stimulated, and one eye open and one eye stimulated. RESULTS: Changes in HbO2 between the rest and stimulation phases were not statistically different between the unilateral and bilateral stimulation conditions with both eyes open. However, HbO2 change was significantly higher in subjects with both eyes open than in subjects with one eye closed (P < 0.001, all comparisons). CONCLUSION: Greater activation of the visual cortex is achieved when subjects are treated with both eyes open as compared to subjects with one eye occluded. From a perspective of functional brain activation, amblyopia treatment administered without occluding the healthy eye may provide the greatest therapeutic benefit.

Is the Cortical Deficit in Amblyopia Due to Reduced Cortical Magnification, Loss of Neural Resolution, or Neural Disorganization?

The neural basis of amblyopia is a matter of debate. The following possibilities have been suggested: loss of foveal cells, reduced cortical magnification, loss of spatial resolution of foveal cells, and topographical disarray in the cellular map. To resolve this we undertook a population receptive field (pRF) functional magnetic resonance imaging analysis in the central field in humans with moderate-to-severe amblyopia. We measured the relationship between averaged pRF size and retinal eccentricity in retinotopic visual areas. Results showed that cortical magnification is normal in the foveal field of strabismic amblyopes. However, the pRF sizes are enlarged for the amblyopic eye. We speculate that the pRF enlargement reflects loss of cellular resolution or an increased cellular positional disarray within the representation of the amblyopic eye. SIGNIFICANCE STATEMENT: The neural basis of amblyopia, a visual deficit affecting 3% of the human population, remains a matter of debate. We undertook the first population receptive field functional magnetic resonance imaging analysis in participants with amblyopia and compared the projections from the amblyopic and fellow normal eye in the visual cortex. The projection from the amblyopic eye was found to have a normal cortical magnification factor, enlarged population receptive field sizes, and topographic disorganization in all early visual areas. This is consistent with an explanation of amblyopia as an immature system with a normal complement of cells whose spatial resolution is reduced and whose topographical map is disordered. This bears upon a number of competing theories for the psychophysical defect and affects future treatment therapies.

The distinct role of NR2B subunit in the enhancement of visual plasticity in adulthood.

BACKGROUND: Experience-dependent plasticity is confined to the critical period of early postnatal life, and declines dramatically thereafter. This attenuation promotes the stabilization of cortical circuits, but also limits functional recovery of several brain diseases. The cognitive functions and synaptic plasticity in the hippocampus and prefrontal cortex are elevated following chronic magnesium treatment. Here, we explored the effect of magnesium treatment on visual plasticity and the potential clinical significance. RESULTS: Visual plasticity in adult mice was dramatically enhanced following magnesium treatment, which was concurrent with an increase in the expression of NR2 subunits of N-methyl-D-aspartate receptors. Blockade of NR2B activity in both the induction and expression periods of plasticity prevented this reinstatement. However, the plasticity restored via a decrease in cortical inhibition was independent on the activation of NR2B, indicating a different underlying mechanism. The functional excitatory synapses on layer 2/3 pyramidal neurons were increased following magnesium supplementation. Moreover, the synaptic and neuronal responses were reminiscent of that within the critical period, and this rejuvenation of adult visual cortex facilitated the recovery of visual functions in amblyopia. CONCLUSIONS: Collectively, our data reveal two distinct mechanisms underlying the restoration of visual plasticity in adulthood, and the rejuvenation of adult visual cortex following magnesium treatment provides a new avenue to develop clinical therapies for adult amblyopia, as well as to explore plasticity-based treatment of other brain diseases, such as stroke and aphasia.

Postnatal visual deprivation in rats regulates several retinal genes and proteins, including differentiation-associated fibroblast growth factor-2.

Little is known about the retinal cellular basis of amblyopia, which is a developmental disease characterized by impaired visual acuity. This study examined the retinal transcripts associated with experimentally induced unilateral amblyopia in rats. Surgical tarsorrhaphy of the eyelids on one side was performed in pups prior to eye opening at postnatal day 14, thereby preventing any visual experience. This condition was maintained for over 2 months, after which electroretinograms (ERGs) were recorded, the retinal ganglion cell (RGC) arrangement and number were determined using neuroanatomical tracing, the retinal transcripts were studied using microarray analysis, regulated mRNAs were confirmed with quantitative reverse-transcriptase PCR, and proteins were stained using Western blotting and immunohistochemistry. An attenuated ERG was found in eyes that were deprived of visual experience. Retrograde neuroanatomical staining disclosed a larger number of RGCs within the retina on the visually deprived side compared to the non-deprived, control side, and a multilayered distribution of RGCs. At the retinomic level, several transcripts associated with retinal differentiation, such as fibroblast growth factor 2 (FGF-2), were either up- or downregulated. Most of the transcripts could be verified at the mRNA level. To unravel the role of a differentiation-associated protein, we tested FGF-2 in dissociated postnatal retinal cell cultures and found that FGF-2 is a potent factor triggering ganglion cell differentiation. The data suggest that visual experience shapes the postnatal retinal differentiation, whereas visual deprivation induces changes at the functional, cellular and molecular levels within the retina.

Blocking PirB up-regulates spines and functional synapses to unlock visual cortical plasticity and facilitate recovery from amblyopia.

During critical periods of development, the brain easily changes in response to environmental stimuli, but this neural plasticity declines by adulthood. By acutely disrupting paired immunoglobulin-like receptor B (PirB) function at specific ages, we show that PirB actively represses neural plasticity throughout life. We disrupted PirB function either by genetically introducing a conditional PirB allele into mice or by minipump infusion of a soluble PirB ectodomain (sPirB) into mouse visual cortex. We found that neural plasticity, as measured by depriving mice of vision in one eye and testing ocular dominance, was enhanced by this treatment both during the critical period and when PirB function was disrupted in adulthood. Acute blockade of PirB triggered the formation of new functional synapses, as indicated by increases in miniature excitatory postsynaptic current (mEPSC) frequency and spine density on dendrites of layer 5 pyramidal neurons. In addition, recovery from amblyopia--the decline in visual acuity and spine density resulting from long-term monocular deprivation--was possible after a 1-week infusion of sPirB after the deprivation period. Thus, neural plasticity in adult visual cortex is actively repressed and can be enhanced by blocking PirB function.

Relationship between monocularly deprivation and amblyopia rats and visual system development.

OBJECTIVE: To explore the changes of lateral geniculate body and visual cortex in monocular strabismus and form deprived amblyopic rat, and visual development plastic stage and visual plasticity in adult rats. METHODS: A total of 60 SD rats ages 13 d were randomly divided into A, B, C three groups with 20 in each group, group A was set as the normal control group without any processing, group B was strabismus amblyopic group, using the unilateral extraocular rectus resection to establish the strabismus amblyopia model, group C was monocular form deprivation amblyopia group using unilateral eyelid edge resection + lid suture. At visual developmental early phase (P25), meta phase (P35), late phase (P45) and adult phase (P120), the lateral geniculate body and visual cortex area 17 of five rats in each group were exacted for C-fos Immunocytochemistry. Neuron morphological changes in lateral geniculate body and visual cortex was observed, the positive neurons differences of C-fos expression induced by light stimulation was measured in each group, and the condition of radiation development of P120 amblyopic adult rats was observed. RESULTS: In groups B and C, C-fos positive cells were significantly lower than the control group at P25 (P<0.05), there was no statistical difference of C-fos protein positive cells between group B and group A (P>0.05), C-fos protein positive cells level of group B was significantly lower than that of group A (P<0.05). The binoculus C-fos protein positive cells level of groups B and C were significantly higher than that of control group at P35, P45 and P120 with statistically significant differences (P<0.05). CONCLUSIONS: The increasing of C-fos expression in geniculate body and visual cortex neurons of adult amblyopia suggests the visual cortex neurons exist a certain degree of visual plasticity.

The expression of vasoactive intestinal polypeptide in visual cortex-17 in normal visual development and formation of anisometropic amblyopia.

AIMS: To document the expression of vasoactive intestinal polypeptide (VIP) in the visual cortex-17 of kittens with anisometropic amblyopia, and to investigate the relationship between VIP and the development of the visual system. METHODS: Sixteen normal kittens (4-wk of age) were randomly divided into two groups: control and amblyopic. Amblyopia was produced by atropinization of one eye in eight kittens. Four (2 normal and 2 amblyopia) kittens were sacrificed at weeks 3, 6, 9, or 12 post-treatment respectively. Expression of VIP-mRNA in the visual cortex-17 was detected through in-situ hybridization. Neurons in the visual cortex were visualized by transmission electron microscopy (TEM). The number of neurons was analyzed via light microscopy (LM). RESULTS: VIP-mRNA expression was increased with age in control kittens but remained nearly static in age-matched anisometropic amblyopic kittens (p < 0.05). The number of VIP-positive cells of amblyopic kittens decreased dramatically when compared to normal age-matched kittens (p < 0.05). The total comparison between different positive ranks suggested a significant difference. The degree of expression between these two groups was significantly different. Ultrastructurally, in the control group, the nuclear membrane of most neurons was discernable and chromatin was evenly distributed within the nucleus. Abundant cytoplasm and tubular-shaped mitochondria were observed. These cells were also rich in Golgi bodies, ribosomes, and endoplasmic reticulum. In amblyopic kittens, nuclei of most neurons were aggregated, the number of ribosomes and Golgi bodies was reduced, mitochondria were swollen, and mitochondrial cristae were shortened or even absent. The endoplasmic reticulum was distended and reduced in magnitude. CONCLUSIONS: VIP appears to play an important role in visual development, and its mRNA expression is affected by visual experiences. Visual dysfunction may down-regulate the expression of VIP-mRNA by impairing the structure and function of the neurons in the visual cortex, finally leading to amblyopia.

L-dopa methyl ester attenuates amblyopia-induced neuronal injury in visual cortex of amblyopic cat.

In the present study, we aimed to assess the potential anti-amblyopic effects of L-dopa methyl ester (LDME) on visual cortex area 17 in an amblyopic feline model induced by monocular vision deprivation. After LDME administration, pathophysiologic and ultrastructural observations were utilized to examine the morphological changes of nerve cells in visual cortex area 17. Dopamine (DA) and its metabolite contents in visual cortex area 17 were investigated through HPLC analysis. Apoptotic cells in visual cortex area 17 were evaluated by TUNEL assay. Additionally, the c-fos expression both at gene and protein levels was assessed using RT-PCR and immunohistochemistry analyses, respectively. The contents of DA and its metabolites were elevated in visual cortex area 17. Neuronal rejuvenation which occurred in visual cortex area 17 was observed through anatomical and physiological assessments. Similarly, TUNEL results showed that neuronal apoptosis was inhibited in the visual cortex of amblyopic cats by both L-dopa and LDME therapies. Meanwhile, the c-fos expression was notably up-regulated at both the mRNA and protein levels by the treatments. These findings suggested that LDME treatment could effectively increase DA and its metabolite contents, and restrain the apoptotic process, as well as elevate the c-fos expression in nerve cells of visual cortex area 17. Taken together, LDME might ameliorate the functional cytoarchitecture in visual cortex area 17 through mechanisms that elevate DA content and increase endogenous c-fos expression, as well as inhibit neuronal lesion in visual cortex tissue.