Form Deprivation Myopia In Guinea Pigs Research Articles

Changes in choroidal hemodynamics of form-deprivation myopia in Guinea pigs

PurposeWe studied changes in the choroid, particularly variation in blood flow, during the development of myopia. The hemodynamic mechanism in play remains unclear. We evaluated blood flow by quantitating indocyanine green (ICG) fluorescence in a guinea pig model of form-deprivation myopia. MethodsGuinea pigs were divided into form-deprivation myopia (FDM) and normal control (NC) groups. Ocular biometric and choroidal hemodynamics parameters were quantitatively derived via ICG imaging, and included the maximal ICG fluorescence intensity (Imax), rising time (Trising), blood flow index (BFI), and mean transit time (MTT). ResultsForm deprivation was associated with significant interocular differences in terms of both refractive error and axial length. ICG fluorescence hemodynamic maps of fundal blood flow and vasculature density were evident. In deprived eyes, the fluorescence signals exhibited significantly longer Trising and MTT but lower Imax and BFI than fellow eyes and NC group. The interocular differences in terms of the ocular biometric and hemodynamic parameters were significantly correlated. Hemodynamic analysis of choriocapillaris lobules revealed weakened fluorescence intensity and prolonged arrival and filling times in deprived eyes. Form deprivation reduced the number of lobulated choriocapillaris structures. ConclusionForm-deprivation myopia triggered changes in the hemodynamic and vascular network structures of the choroid and choriocapillaris. The ICG fluorescence imaging/analysis method provides a unique tool for further myopia research.

Widespread Involvement of Acetylation in the Retinal Metabolism of Form-Deprivation Myopia in Guinea Pigs.

Myopia has become the major cause of visual impairment worldwide. Although the pathogenesis of myopia remains controversial, proteomic studies suggest that dysregulation of retinal metabolism is potentially involved in the pathology of myopia. Lysine acetylation of proteins plays a key role in regulating cellular metabolism, but little is known about its role in the form-deprived myopic retina. Hence, a comprehensive analysis of proteomic and acetylomic changes in the retinas of guinea pigs with form-deprivation myopia was performed. In total, 85 significantly differential proteins and 314 significantly differentially acetylated proteins were identified. Notably, the differentially acetylated proteins were markedly enriched in metabolic pathways such as glycolysis/gluconeogenesis, the pentose phosphate pathway, retinol metabolism, and the HIF-1 signaling pathway. HK2, HKDC1, PKM, LDH, GAPDH, and ENO1 were the key enzymes in these metabolic pathways with decreased acetylation levels in the form-deprivation myopia group. Altered lysine acetylation of key enzymes in the form-deprived myopic retina might affect the dynamic balance of metabolism in the retinal microenvironment by altering their activity. In conclusion, as the first report on the myopic retinal acetylome, this study provides a reliable basis for further studies on myopic retinal acetylation.

Quantitative Assessment of the Choroidal Vessel Diameter during the Recovery of Form-Deprivation Myopia in Guinea Pigs

Purpose The development and recovery (REC) of myopia is associated with changing of choroidal thickness (CT) in the model of guinea pigs. Nitric oxide synthase (NOS) is an enzyme which can affect choroidal vasodilatation. This study wants to investigate the changes of choroidal vessel diameter (CVD) and NOS during the REC of form-deprivation (FD) myopia in guinea pigs. Methods Forty-eight guinea pigs were randomly assigned to the normal control (NC) group, FD group (FD for 21 d), and four REC groups: REC1/2 group (removal the deprivation and re-exposure to the normal environment for 1/2 d), REC1 group (1 d), REC 2 group (2 d), and REC7 group (7 d). CT was measured by optical coherence tomography (OCT), and CVD of foveal choroid was quantitatively assessed on OCT angiography images using MATLAB software at each time point. NOS in choroid was measured using enzyme-linked immunosorbent assays. Measurements were compared between groups and correlations between CT, CVD, and NOS were assessed using regression analyses. Results CVD and CT in FD group were significantly smaller than in NC group (both p < .05), while the NOS significantly larger (p < .001). When deprivation was removed, CVD and NOS were significantly larger and reached a peak in the REC1 group, while CT reached the peak in the REC2 group, then all gradually decreased, and no significant differences were observed in NC and REC7 group (all p > .05). In the REC and NC groups, there was a significant positive correlation between CVD and NOS (p < .001), CVD and CT (p = .0092), but no correlation was found between NOS and CT (p > .05). Conclusions This study indicated that the CVD in guinea pigs could be significantly dilated following myopia REC, and this change coincides with changes in NOS and CT.

Expression of SCO1 and SCO2 after form-deprivation myopia in Guinea pigs.

The retina is a highly energy-consuming tissue associated with visual development, and the reduced quality of retinal imaging can be related to myopia. Synthesis of cytochrome c oxidase 1 (SCO1) and synthesis of cytochrome c oxidase 2 (SCO2) are involved in ATP (adenosine triphosphate) synthesis and energy metabolism. This study aimed to observe the morphologic changes and investigate the expression of SCO1 and SCO2 induced by form-deprivation myopia (FDM) in the retina and sclera of guinea pigs. Thirty-six 3-week-old male guinea pigs were randomly assigned to one of two groups: (1) the model group (n = 18), in which the right eyes were covered by a thin opaque balloon as FDM group, and the left eyes were uncovered and served as the contralateral control group; (2) the blank control group (n = 18), in which bilateral eye received no manipulation. Eyeballs were enucleated for histological analysis. The retina and sclera of the guinea pigs were separated to determine the protein and mRNA expression levels of SCO1 and SCO2, respectively. After four weeks of form deprivation (FD), the refractive degree and axial length increased significantly (P < 0.001). The retinal and scleral tissues were moderately thinner, and the ganglion cells and the cells of inner and outer nuclear layers in the retina became fewer. Compared with the contralateral control group (P < 0.001) and the blank control group (P < 0.001), the collagen content of the sclera became less in the FDM group. The protein and mRNA expression levels of SCO1 and SCO2 in the FDM group were significantly lower than those in the contralateral control group and the blank control group (P < 0.05). The morphologies of the retina and sclera were changed, and the expression of SCO1 and SCO2 at the protein and transcription levels was significantly reduced in the FDM group. Given these changes, SCO1 and SCO2 genes may be involved in myopic progression.

Dynamic Changes of AREG in the Sclera during the Development of Form-Deprivation Myopia in Guinea Pigs

ABSTRACTPurposeTo investigate amphiregulin (AREG) expression in the sclera during the development of form-deprivation myopia (FDM) and after the recovery of FDM in guinea pigs.MethodsSixty-four 2-week-old guinea pigs were randomly divided into the control and FDM groups. The right eyes of animals in FDM group were covered for 2 weeks (2 W) and 4 weeks (4 W), or were covered for 4 weeks and then uncovered for the subsequent 2 weeks (6 W). The diopters and axial lengths (AL) in the right eyes of guinea pigs were measured. Reverse transcription polymerase chain reaction (RT-PCR), immunofluorescence, and western blotting assays were used to detect the relative mRNA and protein expressions of AREG in the sclera of guinea pigs.ResultsBefore masking (0 W), the refraction and AL in the right eyes of guinea pigs in the control and FDM groups did not differ significantly (both p > .05). Myopic shift was induced in guinea pigs with the diopters gradually decreased and AL increased in the FDM group. While no significant difference was found in control group at different time points, the relative AREG mRNA and protein expression levels in the FDM group were significantly increased in 2 W and 4 W and then decreased after 4 weeks of covering followed by uncovering for 2 weeks (all p < .05).ConclusionsAREG was expressed in the sclera of guinea pigs. Moreover, the expression levels of AREG increased during the development of FDM and downregulated after recovery of FDM. Therefore, AREG may be involved in the regulation of scleral remodeling in myopia.

Intravitreal brimonidine inhibits form-deprivation myopia in guinea pigs

BackgroundThe use of ocular hypotensive drugs has been reported to attenuate myopia progression. This study explores whether brimonidine can slow myopia progression in the guinea pig form-deprivation (FD) model.MethodsThree-week-old pigmented male guinea pigs (Cavia porcellus) underwent monocular FD and were treated with 3 different methods of brimonidine administration (eye drops, subconjunctival or intravitreal injections). Four different concentrations of brimonidine were tested for intravitreal injection (2 μg/μL, 4 μg/μL, 20 μg/μL, 40 μg/μL). All treatments continued for a period of 21 days. Tonometry, retinoscopy, and A-scan ultrasonography were used to monitor intraocular pressure (IOP), refractive error and axial length (AL), respectively. On day 21, guinea pigs were sacrificed for RNA sequencing (RNA-seq) to screen for associated transcriptomic changes.ResultsThe myopia model was successfully established in FD animals (control eye vs. FD eye, respectively: refraction at day 20, 0.97 ± 0.18 D vs. − 0.13 ± 0.38 D, F = 6.921, P = 0.02; AL difference between day 0 and day 21, 0.29 ± 0.04 mm vs. 0.45 ± 0.03 mm, F = 11.655, P = 0.004). Among the 3 different brimonidine administration methods, intravitreal injection was the most effective in slowing myopia progression, and 4 μg/μL was the most effective among the four different concentrations of brimonidine intravitreal injection tested. The AL and the refraction of the brimonidine intravitreal injection group was significantly shorter or more hyperopic than those of other 2 groups. Four μg/μL produced the smallest difference in AL and spherical equivalent difference values. FD treatment significantly increased the IOP. IOP was significantly lower at 1 day after intravitreal injections which was the lowest in FD eye of intravitreal injection of brimonidine. At day 21, gene expression analyses using RNA-seq showed upregulation of Col1a1 and Mmp2 expression levels by intravitreal brimonidine.ConclusionsAmong the 3 different administration methods, intravitreal injection of brimonidine was the most effective in slowing myopia progression in the FD guinea pig model. Intravitreal brimonidine at 4 μg/μL significantly reduced the development of FD myopia in guinea pigs. Expression levels of the Col1a1 and Mmp2 genes were significantly increased in the retinal tissues of the FD-Inj-Br group.

The Role of C-Jun N-terminal Kinase-1 in Controlling Aquaporin-1 and Choroidal Thickness during Recovery from Form-deprivation Myopia in Guinea Pigs

ABSTRACT Purpose The development and recovery(REC) of myopia is associated with changing of choroidal thickness(CT) in model of guinea pigs. Aquaporin-1 (AQP-1) is related to the changes of CT during the recovery from myopia, but the corresponding signaling pathway has not been clarified. This study aimed to investigate the effect of JNK1 on CT/AQP-1 and the recovery from myopia. Materials and Methods According to the different single intravitreal injections in eyes that underwent form deprivation for 21 days, guinea pigs were divided into four groups: the REC group, the REC+anisomycin (REC-AN, agonist for JNK1, 0.2 nmol) group, the REC+SP600125 (REC-SP, inhibitor for JNK1, 0.2 nmol) group, and the REC+dimethyl sulfoxide (REC-DM) group. Each group was divided into three subgroups based on the duration of the form deprivation: 3 days (d), 7 d and 10 d. All animals underwent biometric measurements (refractive error, axial length (AL), and CT), and the protein expression of AQP-1 and p-JNK1 in the choroid was also measured. Results In REC and REC-DM groups, significant differences in CT/refractive error/AL/p-JNK1 or AQP-1 were only found in the 3d group compared with normal control (NC) group (all p < .05). In REC-AN group, CT/p-JNK1 or AQP-1 in 3d group was significantly higher than that in other 3d groups (all p < .05), but no significant difference in refractive error or AL was found compared with NC group at three time points (all p > .05). In REC-SP group, a significant difference in refractive error/CT/p-JNK1 or AQP1 was found in 3d/7d group compared with NC group (all p < .05), but AL was only found in 3d groups (p = .001). Conclusions Changes in JNK1 phosphorylation can regulate AQP-1 and CT during the recovery from myopia and the recovery time. Thus, JNK1 may be a potential therapeutic target for preventing/treating myopia.

DNA methylation and mRNA expression of IGF-1 and MMP-2 after form-deprivation myopia in guinea pigs.

The molecular mechanism of form-deprivation myopia is unclear. This study was aimed to investigate the roles of scleral DNA methylation and mRNA expression of IGF-1 and MMP-2 in a guinea pig model of form-deprivation myopia. Seventy 2-week-old male guinea pigs were assigned to three groups: (1) zero week group that was used to collect baseline data; (2) monocular deprivation treatment (MDT) group, in which a thin slice of opaque latex glove was placed over the right eyes of the animals for four weeks, and the left eyes were untreated and served as the monocular contralateral control (MCC) group; (3) control group (CG), in which the animals grew four weeks, but received no manipulation. Animals in each group were evenly divided for DNA methylation assay and quantitative PCR (qPCR). After eye enucleation, the sclerae were harvested for DNA methylation assay and qPCR. The DNA methylation pattern in the promoter and exon regions of IGF-1 and MMP-2, along with the mRNA expression level of them, were determined by base-specific cleavage and mass spectrometry and qPCR, respectively. After four weeks of form-deprivation, DNA methylation at 4/8 cytosine-guanine sites in the IGF-1 promoter was significantly lower in the MDT eyes than in the MCC or CG eyes. In addition, the level of IGF-1 mRNA was moderately higher in MDT eyes compared to the MCC eyes and CG eyes. DNA methylation at 4/14 cytosine-guanine sites in the MMP-2 gene was very low, and no significant change was observed between the MDT eyes and the MCC or CG ones. However, the level of MMP-2 mRNA in MDT eyes was significant higher compared with MCC eyes and CG eyes, with an increase of 217% and 222%, respectively. In our guinea pig model of form-deprivation myopia, the methylation of four cytosine-guanine sites in the IGF-1 gene promoter was significantly lower in the sclera after four weeks of MDT, and the transcription level of scleral IGF-1 was moderately higher. Hence, the IGF-1 gene methylation might play a role in the pathogenesis of form-deprivation myopia in guinea pigs. The level of MMP-2 mRNA in the sclera of MDT eyes was significantly higher, but not regulated by the methylation pathway, as the methylation status of MMP-2 was unchanged.

Relationship between Aquaporin-1 Protein Expression and Choroidal Thickness during the Recovery of Form-deprivation Myopia in Guinea Pigs

ABSTRACTPurpose: The purpose of this study was to investigate the relationship between aquaporin-1 (AQP-1) protein expression in the choroid and choroid thickness (CT) during the recovery of form-deprivation (FD) myopia in guinea pigs.Materials and Methods: Seventy-two guinea pigs were randomly assigned to the normal control (NC) group, FD 21 group (animals wore a latex facemask in the right eye for 21 days to induce FD myopia) and four recovery (REC) groups. Guinea pigs in the REC groups also wore the facemask for 21 days to induce myopia; then, the facemask was removed, and the eye was re-exposed to the normal environment for 12 hours (REC ½ group), 1 day (REC 1 group), 2 days (REC 2 group), and 7 days (REC 7 group). All animals underwent biometric measurements (refraction, axial length, and CT), and the protein expression of AQP-1 in the choroid was determined using western blotting.Results: The protein expression of AQP-1 and CT were significantly decreased in the FD 21 group as compared with those in the NC group (p = .007 and p < .001). Both AQP-1 protein expression and CT gradually increased and peaked in the REC 2 group. Additionally, there were significant differences in AQP-1 protein expression and CT between the REC 2 group and all other groups (all p < .05). We observed a complete recovery in the in REC 7 group as compared with the NC group (p > .05). AQP-1 protein expression was significantly associated with CT (p = .001) in all groups; however, there was a significant negative correlation (p = .029) between AQP-1 protein expression and axial length in the REC groups.Conclusions: AQP-1 protein expression in the choroid was upregulated following recovery of FD myopia in guinea pigs, and these changes correlated with alterations in CT and axial length.

The Effects of High Lighting on the Development of Form-Deprivation Myopia in Guinea Pigs.

To investigate the effects of high ambient lighting on refraction and ocular biometry in guinea pig models of form-deprivation myopia (FDM). Forty 3-week-old guinea pigs were randomly assigned to groups exposed to either high light (HL, 10,000 lux) or normal light (NL, 500 lux) with normal vision or form deprivation. Throughout the 10-week rearing period, animals were exposed to high light or normal light for 12 hours with a 12-hour light/dark cycle. Refraction, axial length (AL), and radius of corneal curvature (CCR) were measured by cycloplegic retinoscopy, A-scan ultrasonography, and keratometer, respectively. At the end of treatment, form-deprived eyes under high ambient lighting exhibited more hyperopic refraction and shorter AL than those under normal ambient lighting (2.06 ± 1.68 diopters [D; mean ± SD] vs. -0.59 ± 1.56 D, P < 0.001; 8.36 ± 0.13 mm vs. 8.56 ± 0.16 mm, P < 0.001). Deprived eyes under high ambient lighting were relatively more myopic than their contralateral control eyes at the end of treatment (2.06 ± 1.68 D vs. 5.44 ± 0.66 D, P < 0.001). High lighting induced a significant hyperopic shift in normal eyes after 4 weeks of exposure. There were no significant differences in CCR between eyes exposed to high and normal light, nor between deprived eyes and contralateral eyes. High ambient lighting could retard, but not fully inhibit, the development of FDM. High light levels contributed to a greater hyperopic shift in normal eyes during the first 4 weeks of treatment. Corneal curvature was unaffected by either high ambient lighting or form deprivation.

Effect of Bilberry Extract on Development of Form-Deprivation Myopia in the Guinea Pig.

To investigate the effects of bilberry extract, Difrarel, on form-deprivation myopia in guinea pigs. Thirty healthy pigmented guinea pigs, at the age of 3 weeks, were randomly assigned to 3 groups: 2 groups receiving daily orally administered Difrarel (300 mg/kg) either from the outset for 4 or 2 weeks after form deprivation and a control group. Form deprivation was induced with translucent eye patches on the right eye and lasted for 4 weeks. Another 12 guinea pigs without the form deprivation were assigned into 2 groups for oral administration of Difrarel and saline for 4 weeks. The refractive errors and axial length were measured in both eyes, and the posterior sclera was removed and assayed to evaluate the expressions of matrix metalloproteinase 2 (MMP2) and collagen I by a quantitative polymerase chain reaction and Western blot. Oral administration of Difrarel for 4 weeks was found to inhibit the axial elongation and decrease the myopic shift of refractive errors in the form-deprived eye. Upregulation of MMP2 and degradation of collagen I in the sclera of form-deprived eye were observed, and these were prevented by orally administered Difrarel. Moreover, oral administration of Difrarel had no effects on axial length, refractive errors, and the expression of MMP2 in the normal eye. This work demonstrates that orally administered Difrarel has an effect against form-deprivation myopia and raises the possibility that bilberry extracts or specific components might represent a useful treatment for preventing the progression of myopia in clinical practice.

The Role of cGMP in Ocular Growth and the Development of Form-Deprivation Myopia in Guinea Pigs

Development of myopia is associated with remodeling of the sclera, a tissue composed principally of collagen. Cyclic guanosine monophosphate (cGMP) regulates collagen synthesis in several organs; therefore, we investigated the effects of soluble guanylyl cyclase (sGC) stimulation and inhibition on refraction and ocular growth in guinea pigs under normal and form-deprived (FD) conditions. Retinal and scleral cGMP concentrations were measured in normal and monocularly FD guinea pigs at 2 days, 1 week, and 2 weeks of form deprivation and following 2 days recovery. Stimulation of sGC by BAY41-2272 and inhibition by NS-2028 were achieved by daily subconjunctival injection in normal and FD eyes. Refraction and axial parameters were measured at the commencement, middle, and cessation of the experiment. cGMP levels were also determined at the end of the experiment. Retinal and scleral cGMP concentrations increased in FD eyes from 2 days to 2 weeks (P ≤ 0.029). Levels decreased after 2 days of recovery (P ≤ 0.003). Daily injections of BAY41-2272 induced a myopic shift (P ≤ 0.001) and ocular elongation (P ≤ 0.01) in normal animals, but did not alter myopia in FD eyes (P > 0.05). In contrast, daily injections of NS-2028 partially reduced myopic shifts (P ≤ 0.012) and ocular elongation (P ≤ 0.015) induced by form deprivation, but did not affect ocular growth and refraction in normal eyes (P > 0.05). Retinal and scleral cGMP levels were increased by BAY41-2272 in normal eyes and decreased by NS-2028 in FD eyes. Changes in cGMP signaling contribute to myopic development. Thus, cGMP may be a potential therapeutic target for preventing/treating myopia.

Dynamic change of collagen typeIand fibronectin in posterior sclera of form-deprivation myopia after TIMP-2 gene transfect in guinea pig

Background The domestic and international researches discovered that many proteins and enzymes of the extracellular matrix (ECM) participate in the sclera remodeling by affecting the collagen typeⅠand fibronectin.Objective This study was to investigate the effect of matrixmetalloproteinase-2 (TIMP-2) on expression of collagen typeⅠand fibronectin of ECM in the posterior sclera by injecting liposomes containing tissue inhibitor of TIMP-2 gene into suprachoroidal space of the form-deprivation myopia in guinea pig.Methods Form-deprivation myopia was induced by translucent goggles in 36 clean guinea pig for 2 weeks.Then the animals were randomly assigned to TIMP-2 group,empty plasmid group,saline group and 12 for each group.Liposomes of 5μl containing TIMP-2 gene,empty plasmid and saline were suprachoroidally injected in the right eye respectively,and the left eyes without any treatment were used as self-control group.Other 12 matched guinea pigs only covered the right eyes through out the experimental duration as model control group.The guinea pigs were sacrificed and the posterior sclera tissue of the eyeballs were collected at 2,7 and 14 days after injection of drug.The expressions of collagen typeⅠmRNA and fibronectin mRNA were detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR).This study followed the Regulation for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.Results The expression level of collagen type Ⅰ mRNA in the posterior sclera of guinea pig was lower but that of fibronectin mRNA was higher in TIMP-2 group than self-control group,showing significant differences between them (P＜0.05).The expression level of collagen type Ⅰ mRNA in the posterior scleral tissue began to increase from the 2nd day after drug injection and was obviously elevated at the 7th day and then gradually decreased at the 14th day.However,the expression level of fibronectin mRNA in the posterior scleral tissue showed the opposite pattern.The expression levels of collagen typeⅠmRNA and fibronectin mRNA at the 7th after drug injection were significantly lower than that at the 2nd day or 14th day (P＜0.01).Conclusion Suprachoroidal injection of TIMP-2 in form-deprivation myopia could up-regulate the expression of collagen typeⅠmRNA and down-regulate the expression of fibronectin mRNA in the posterior scleral tissue.It may slow down the sclera remodeling of form-deprivation myopia in guinea pig in the early stage. Key words: Form-deprivation myopia; Tissue inhibitor of matrixmetalloproteinase-2; Extracellular matrix; Collagen type Ⅰ; Fibronectin

Levodopa Inhibits the Development of Form-Deprivation Myopia in Guinea Pigs

It has been shown that visual deprivation leads to a myopic refractive error and also reduces the retinal concentration of dopamine. Exogenously 3,4-dihydroxy-L-phenylalanine (levodopa, L-DOPA) can be converted into dopamine in vivo, which safely and effectively treats Parkinson disease. Moreover, L-DOPA was also used in the treatment of amblyopia in clinical studies. However, the effect of L-DOPA on the development of myopia has not been studied. The aim of this study was to investigate whether intraperitoneal injection of L-DOPA could inhibit form-deprivation myopia in guinea pigs and to explore a new strategy for drug treatment of myopia. Sixty guinea pigs, at age of 4 weeks, were randomly divided into six groups: normal control, L-DOPA group, saline group, deprived group, deprived plus L-DOPA group, and deprived plus saline group. Form deprivation was induced with translucent eye shields on the right eye and lasted for 10 days. L-DOPA was injected intraperitoneally into the guinea pig once a day. The corneal radius of curvature, refraction, and axial length were measured in all animals. Subsequently, retinal dopamine content was evaluated by high-performance liquid chromatography with electrochemical detection. Ten days of eye occlusion caused the form-deprived eyes to elongate and become myopic, and retinal dopamine content to decrease, but the corneal radius of curvature was not affected. Repeated intraperitoneal injection of L-DOPA could inhibit the myopic shift (from -3.62 +/- 0.98 D to -1.50 +/- 0.38 D; p < 0.001) due to goggles occluding and compensate retinal dopamine (from 0.65 +/- 0.10 ng to 1.33 +/- 0.23 ng; p < 0.001). Administration of L-DOPA to the unoccluded animals had no effect on its ocular refraction. There was no effect of intraperitoneal saline on the ocular refractive state and retinal dopamine. Systemic L-DOPA was partly effective in this guinea pig model and, therefore, is worth testing for effectiveness in progressing human myopes.

Time-course of changes to nitric oxide signaling pathways in form-deprivation myopia in guinea pigs

The aim of this study was to investigate the time-course change of nitric oxide synthase (NOS) activity and cyclic GMP (cGMP) concentration in the posterior retina, choroid and sclera after differing periods of form-deprivation in guinea pigs. Three groups of guinea pigs were subjected to monocular FD for 7, 14 or 21 days. NOS activity and cGMP concentrations in ocular tissues of FD eyes and control eyes were analyzed by radioimmunoassay. The presence of NOS isoforms was detected by immunohistochemistry. Guinea pigs presented with considerable myopia after 14 days of FD. Retinal NOS activity in the FD group was lower than in the control group after 7 days of FD and was higher than in the control group after 14 and 21 days of FD. The choroidal and scleral NOS activities in the FD groups were higher than in the control groups after 21 days. The cGMP concentrations in the FD groups were higher than in the control groups at 21 days of the retinal, choroidal, and scleral tissues. Furthermore, the retinal cGMP concentration in the FD group was also significantly elevated at 14 days relative to the control group. We detected expression of three NOS isoforms in guinea pig ocular tissues. Our main observations were a change in NOS activity and an up-regulation in cGMP concentrations in posterior ocular tissues during the development of myopia. The function of elevated NOS activity may be mediated by cGMP.