Serial magnetic resonance imaging study of posterior cruciate ligament reconstruction or augmentation using hamstring tendons.

PURPOSE: The purpose of this study was to analyze serial changes in the magnetic resonance imaging (MRI) signals of autograft hamstrings single bundle posterior cruciate ligament (PCL) reconstruction and the effects of remnant preservation (augmentation). MATERIAL AND METHODS: Twenty-two isolated PCL injuries were arthroscopically reconstructed or augmented with hamstring tendons. MRI scans were obtained at 3, 6, and 12 months, and prior to the second-look arthroscopy (average 20.7 months). The patients were divided into 2 groups by remnant preservation: five PCL reconstructions after PCL remnant resection (Group Rec) (23%), and 17 reconstructions preserving the remnant (Group Aug) (77%). The 22 patients were also divided in two groups depending on the location of the PCL tear. There were 9 knees with proximal tear (Type P) (41%) and 13 knees with distal tear (Type D) (59%). The signal intensity and fiber continuity of 4 zones (proximal, middle, distal intra-articular and tibial tunnel zones) were evaluated by the Mariani score. RESULTS: The average MRI evaluation score gradually increased from 6 months through the final MRI. The intra-articular part of the graft exhibited slower maturation (12 months - final scan) as compared with the tibial tunnel (6-12 months). The distal zone underwent better maturation than the proximal or middle zones at all points. In the proximal zone, the score for Group Aug was significantly higher than Group Rec. In the proximal zone, the Type D score with a proximally-preserved remnant was significantly higher than Type P without a proximal remnant. CONCLUSIONS: The hamstring tendons require more than 1 year to achieve low-signal intensity. PCL remnant has a beneficial effect on the maturation of the hamstring graft. LEVEL OF EVIDENCE IV: therapeutic case series.

Association study of genetic variants on chromosome 7q31 with susceptibility to normal tension glaucoma in a Japanese population.

The caveolin 1 to caveolin 2 (CAV1-CAV2) gene region on chromosome 7q31 has been reported to be associated with susceptibility to primary open angle glaucoma (POAG) and normal tension glaucoma (NTG) in previous studies. We investigated whether genetic variants in the CAV1-CAV2 region are associated with NTG in Japanese patients. Two hundred and ninety-two Japanese patients with NTG and 352 Japanese healthy controls were recruited. We genotyped three single-nucleotide polymorphisms; that is, rs1052990, rs4236601, and rs7795356, in the CAV1-CAV2 gene region and assessed the allelic diversity among cases and controls. The frequency of the minor allele (G) of rs1052990 was significantly decreased in NTG cases compared with controls (P=0.014, OR=0.71), whereas NTG or POAG cases had a significantly higher frequency of the allele than controls in previous studies. Conversely, rs7795356 did not show any significant association with NTG cases, and rs4236601 was monomorphic in the Japanese study population. Our findings did not correspond with previous positive results, suggesting that CAV1-CAV2 variants studied in the present study are not important risk factors for NTG susceptibility in all populations. Further studies are needed to elucidate the possible contribution of the CAV1-CAV2 region to the development of glaucoma.

Comparing approaches to screening for angle closure in older Chinese adults.

AIMS: Primary angle-closure glaucoma is expected to account for nearly 50% of bilateral glaucoma blindness by 2020. This study was conducted to assess the performance of the scanning peripheral anterior chamber depth analyzer (SPAC) and limbal anterior chamber depth (LACD) as screening methods for angle closure. METHODS: This study assessed two clinical populations to compare SPAC, LACD, and gonioscopy: the Zhongshan Angle-closure Prevention Trial, from which 370 patients were eligible as closed-angle participants and the Liwan Eye Study, from which 72 patients were selected as open-angle controls. Eligible participants were assessed by SPAC, LACD, and gonioscopy. RESULTS: Angle status was defined by gonioscopy. Area under the receiver operating characteristic curve (AUROC) for SPAC was 0.92 (0.89-0.95) whereas AUROC for LACD was 0.94 (0.92-0.97). Using conventional cutoff points, sensitivity/specificity was 93.0%/70.8% for SPAC and 94.1%/87.5% for LACD. Sequential testing using both SPAC and LACD increased the specificity to 94.4% and decreased the sensitivity to 87.0%. CONCLUSION: SPAC has significantly lower specificity than LACD measurement using conventional cutoffs but interpretation of the findings can be performed by modestly trained personnel.

A remote operating slit lamp microscope system. Development and its utility in ophthalmologic examinations.

OBJECTIVES: To develop a remote-operating slit lamp microscope system (the remote slit lamp) as the core for highly specialized ophthalmology diagnoses, and to compare the utility of this system with the conventional slit lamp microscope system (the conventional slit lamp) in making a diagnosis. METHODS: The remote slit lamp system was developed. Three factors were evaluated in comparison to the conventional slit lamp. The ability to acquire skills was investigated using a task loading system among specialists and residents in ophthalmology. Participants repeated a task up to ten times and the time required for each task was analyzed. The consistency of the two systems in making a diagnosis was investigated using eyes of patients with ocular diseases as well as healthy volunteers. RESULTS: The remote slit lamp is composed of a patient's unit and ophthalmologist's unit connected by high-speed internet. The two units share images acquired by the slit lamp in addition to the images and voices of patients and ophthalmologists. Both ophthalmology specialists and residents could minimize the completion times after several trials. The remote slit lamp took more time than the conventional slit lamp. Both systems showed a high consistency in evaluations among eyes with healthy eyes or those with ocular diseases. CONCLUSIONS: The remote slit lamp has a similar diagnostic ability, but required more examination time in comparison to the conventional slit lamp. The currently developed remote slit lamp has the potential to be employed for tele-medicine purposes in the field of ophthalmology.

Investigation of the association between the GLC3A locus and normal tension glaucoma in Japanese patients by microsatellite analysis.

PURPOSE: To investigate whether the GLC3A locus harboring the CYP1B1 gene is associated with normal tension glaucoma (NTG) in Japanese patients. MATERIALS AND METHODS: One hundred forty-two Japanese patients with NTG and 101 Japanese healthy controls were recruited. Patients exhibiting a comparatively early onset were selected as this suggests that genetic factors may show stronger involvement. Genotyping and assessment of allelic diversity was performed on 13 highly polymorphic microsatellite markers in and around the GLC3A locus. RESULTS: There were decreased frequencies of the 444 allele of D2S0416i and the 258 allele of D2S0425i in cases compared to controls (P = 0.022 and P = 0.034, respectively). However, this statistical significance disappeared when corrected (Pc > 0.05). We did not find any significant association between the remaining 11 microsatellite markers, including D2S177, which may be associated with CYP1B1, and NTG (P > 0.05). CONCLUSIONS: Our study showed no association between the GLCA3 locus and NTG, suggesting that the CYP1B1 gene, which is reportedly involved in a range of glaucoma phenotypes, may not be an associated factor in the pathogenesis of NTG.

Microsatellite analysis of the GLC1B locus on chromosome 2 points to NCK2 as a new candidate gene for normal tension glaucoma.

AIMS: The aim of this study was to investigate the association between normal tension glaucoma and the candidate disease locus glaucoma 1, open angle, B (GLC1B) on chromosome 2. There are many reports describing the results of association or linkage studies for primary open angle glaucoma (POAG), with GLC1B as one of the loci associated with normal or moderately elevated intraocular pressure. However, there are few reports about the association of genes or defined genomic regions with normal tension glaucoma, which is the leading type of glaucoma in Japan. The GLC1B locus is hypothesized to be a causative region for normal tension glaucoma. METHODS: Genomic DNA was extracted from whole blood of normal tension glaucoma (n = 143) and healthy controls (n = 103) of Japanese origin. RESULTS: Fifteen microsatellite markers within and/or near to the GLC1B locus were genotyped, and their association with normal tension glaucoma was analysed. Two markers D2S2264 and D2S176 had significant positive associations. CONCLUSION: The D2S176 marker had the strongest significant association and it is located 24 kb from the nearest gene NCK2, which now becomes an important new candidate gene for future studies of its association with normal tension glaucoma.

Anthraquinone polyamines: novel channel blockers of N-methyl-D-aspartate receptors.

Polyamines, in particular spermine, as well as some natural and synthetic polyamine derivatives have been found to be blockers of N-methyl-D-aspartate receptors. We developed novel, polyamine-based channel blockers to analyze the structure of NMDA receptors. Anthraquinone polyamines block NMDA receptors with some selectivity compared to other glutamate receptors. Results using mutant NR1 and NR2 subunits identified amino acid residues that influence blockade by anthraquinone polyamines. The head group (anthraquinone) may be positioned at the selectivity filter/narrowest constriction of the channel and the polyamine tail penetrates this constriction into the inner vestibule below the level of the selectivity filter. The results are consistent with other work showing that NR1 (Asn616) and NR2B (Asn616), but not NR2B (Asn615), make the narrowest constriction of NMDA channel, and that the M3 segments from the two subunits, which form the outer vestibule, are likely staggered relative to each other in the vertical axis of the channel.

Polyamines in renal failure.

The levels of polyamines (putrescine, spermidine and spermine) and polyamine oxidase in plasma of patients with chronic renal failure were determined. The level of putrescine was increased but the level of spermine was decreased in the plasma of these patients. The patients also had increased plasma polyamine oxidase activity leading to increased degradation of spermine. As acrolein was a major toxic compound produced from spermine by polyamine oxidase, the levels of free and protein-conjugated acrolein in plasma were also measured. Acrolein levels were enhanced in plasma of patients with chronic renal failure. The accumulated acrolein found as protein conjugates was equivalent to 170 microM, which was about 5-fold higher than in plasma of normal subjects. It was found that acrolein is mainly produced by spermine oxidase in plasma. An increase in putrescine, spermine oxidase and acrolein in plasma was observed in all cases such as diabetic nephropathy, chronic glomerulonephritis and nephrosclerosis. After patients with chronic renal failure had undergone hemodialysis, their levels of plasma polyamines, spermine oxidase and acrolein returned towards normal. It is likely that acrolein produced from spermine accumulates in the blood due to decreased excretion into urine and may function as a uremic "toxin".

Effect of non-steroidal anti-inflammatory ophthalmic solution on intraocular pressure reduction by latanoprost in patients with primary open angle glaucoma or ocular hypertension.

AIM: To investigate the effects of a non-steroidal anti-inflammatory drug (NSAID) ophthalmic solution on latanoprost induced intraocular pressure (IOP) reduction in glaucoma patients. METHODS: Examination was conducted on 16 eyes of 16 glaucoma patients who had been given only latanoprost for at least 6 weeks. The NSAID ophthalmic solution, sodium 2-amino-3-(4-bromobenzoyl) phenylacetate sesquihydrate, was additionally given for 12 weeks into one eye (NSAID group), while sodium hyaluronic acid ophthalmic solution was administered into the other eye (control group) in a double masked fashion. The IOP measurement was performed before the start of additional administration of ophthalmic solutions, 2, 4, 6, 8, 10, and 12 weeks after the start of additional administration, and 2, 4, and 6 weeks after discontinuing additional administration. RESULTS: No significant difference was observed in the IOPs before additional administration of ophthalmic solution between the NSAID group and the control group. Following the additional administration of ophthalmic solution, IOP in the NSAID group was consistently higher than that in the control group, and a maximum difference in IOP between the two groups was 1.08 (SD 1.75) mm Hg (p = 0.03). This trend was observed even after additional administration was discontinued. CONCLUSION: NSAID ophthalmic solution may partly affect IOP reduction by latanoprost.

Finding cases of angle-closure glaucoma in clinic setting using a newly developed instrument.

PURPOSE: To validate the applicability of a newly developed, noncontact scanning peripheral anterior chamber depth analyzer (SPAC) for screening eyes at the risk of angle-closure glaucoma (ACG). SUBJECTS AND METHODS: All glaucoma patients who visited the University of Yamanashi Hospital from February through May 2003 were enrolled, except those with aphakic eye or pseudophakic eye. Of the 552 enrolled patients, 48 with ACG or narrow angles requiring laser iridotomy (LI) were categorized as patients with high-risk ACG eyes, and those with open angle were categorized as patients with control eyes. In all, 20 patients with ACG or narrow angles requiring prophylactic LI, who were followed up by an independent private ophthalmic clinic, were enrolled for threshold analysis. Nonophthalmologists measured anterior chamber depth and the averaged values of three measurements were employed for analysis. Threshold analysis and discriminant analysis were employed for determining the sensitivity and specificity of SPAC for diagnosing eyes with high-risk ACG. RESULTS: SPAC distinguished well the high-risk ACG eyes from the control eyes, and one of the most useful criteria for screening is as follows: any of the four measured points should exceed 95% confidence interval, and sensitivity and specificity should be 97.6 and 83.5%, respectively. CONCLUSION: SPAC is thought to be useful for detecting eyes at the risk of ACG by nonophthalmologists.

Usefulness of peripheral anterior chamber depth assessment in glaucoma screening.

PURPOSE: This study was conducted to determine the usefulness of peripheral anterior chamber depth assessment in angle-closure glaucoma (ACG) screening in Japanese subjects. SUBJECTS AND METHODS: The subjects were 14,779 adults 40 years old or older. Eyes having peripheral anterior chamber depth that is 1/4 the peripheral corneal thickness (van Herick's classification: grade 2) and less than 1/4 the peripheral corneal thickness (van Herick's classification: grade 1) were extracted as narrow angle eyes, and those eyes were further examined. RESULTS: Of 14,779 subjects, 923 eyes of 505 subjects were diagnosed as narrow angle eyes (3.4%). Narrow angle eyes were observed in 4.9% of female subjects and 1.9% of male subjects, indicating a significantly higher frequency in women. The percentage of narrow angle eyes increased with age. Among the narrow angle eyes, 61 eyes of 32 subjects were diagnosed with ACG suspect (6.5%). In contrast to the frequency of ACG suspect in eyes classified as grade 1, according to van Herick's classification, being 17.9%, that in eyes classified as grade 2 was significantly lower at 5.6%. CONCLUSION: Since the incidence of ACG suspect increases as the peripheral anterior chamber depth decreases, caution for the peripheral anterior chamber depth is required for the ACG screening.

A newly developed peripheral anterior chamber depth analysis system: principle, accuracy, and reproducibility.

AIM: To develop a new, non-contact system for measuring anterior chamber depth (ACD) quantitatively, and to investigate its accuracy as well as interobserver and intraobserver reproducibility. METHODS: The system scanned the ACD from the optical axis to the limbus in approximately 0.5 second and took 21 consecutive slit lamp images at 0.4 mm intervals. A computer installed program automatically evaluated the ACD, central corneal thickness (CT), and corneal radius of curvature (CRC) instantly. A dummy eye was used for investigating measurement accuracy. The effects of CT and CRC on the measurement results were examined using a computer simulation model to minimise measurement errors. Three examiners measured the ACD in 10 normal eyes, and interobserver and intraobserver reproducibility was analysed. RESULTS: The ACD values measured by this system were very similar to theoretical values. Increase of CRC and decrease in CT decreased ACD and vice versa. Data calibration using evaluated CT and CRC successfully reduced measurement errors. Intraobserver and interobserver variations were small. Their coefficient variation values were 7.4% (SD 2.3%) and 6.7% (0.7%), and these values tended to increase along the distance from the optical axis. CONCLUSION: The current system can measure ACD with high accuracy as well as high intraobserver and interobserver reproducibility. It has potential use in measuring ACD quantitatively and screening subjects with narrow angle.

Quantitative evaluation of changes in anterior segment biometry by peripheral laser iridotomy using newly developed scanning peripheral anterior chamber depth analyser.

AIM: Using the newly developed scanning peripheral anterior chamber depth analyser (SPAC), the effects of peripheral laser iridotomy (PLI) on peripheral anterior chamber depth (PACD) were determined quantitatively as was the association between PACD and chronic elevation of intraocular pressure (IOP) after PLI. METHODS: 16 eyes of 15 patients with acute primary angle closure glaucoma (PACG) attack, 14 eyes of 14 patients with narrow angle and PACG attack in their fellow eyes, and 13 eyes of seven patients with chronic angle closure glaucoma (CACG) were enrolled. The SPAC scanned the anterior ocular segment from the optical axis to the limbus and took 21 consecutive slit lamp images at 0.4 mm intervals. A computer installed program automatically evaluated the PACD and the averaged values of three measurements were employed for analysis. RESULTS: PLI significantly increased PACD and changed the iris contour from convex to flat or concave in all the enrolled eyes. The extent of the PLI induced PACD increase was enhanced with increasing distance from the optical axis. Comparing PACDs after PLI, eyes that received prophylactic PLI showed the greatest extent of PLI induced PACD increase, followed by eyes with CACG and eyes with PACG attack. The PACD of eyes with PACG attack was almost the same as that of the fellow eyes of PACG attack before prophylactic PLI. Eyes with PACG attack showed poorer IOP control after PLI than eyes with narrow angle and CACG with PLI. CONCLUSIONS: PLI significantly increases PACD and the small PLI induced opening of PACD may contribute to chronic IOP elevation after PLI.

Comparison of iridial pigmentation between latanoprost and isopropyl unoprostone: a long term prospective comparative study.

AIM: To compare incidence of iridial pigmentation prospectively induced by long term treatment with latanoprost and isopropyl unoprostone (hereafter, unoprostone) in Japanese patients with glaucoma. METHODS: Patients with glaucoma treated with prostaglandin (PG) related ophthalmic solutions were sequentially enrolled. Patients treated for more than 30 months with PG related ophthalmic solutions were subjected to analysis. The entry criteria were no history of intraocular surgery, laser iridotomy, and/or laser trabeculoplasty within 12 months before and after the enrolment; and no history of uveitis; no changes in antiglaucoma drugs within 6 months before and after the enrolment. Photographs of the irides were taken under the same conditions and three glaucoma specialists evaluated the iridial pigmentation with masking of patient information. The correlation of iridial pigmentation with the background factors and the reduction of intraocular pressure (IOP) before and after the treatment were investigated. RESULTS: 48 eyes in 48 patients satisfied the enrolment criteria (25 eyes in the latanoprost group, 23 eyes in the unoprostone group). At the end of the follow up period, iridial pigmentation was present in 15 patients (60.0%) in the latanoprost group and seven patients (30.4%) in the unoprostone group. The correlation between development of iridial pigmentation and age, sex, concurrent use of other ophthalmic solutions, and IOP reduction was not significant. CONCLUSIONS: The incidence of iridial pigmentation induced by latanoprost or unoprostone is high in the case of long term treatment. Iridial pigmentation did not affect PG related ophthalmic solution induced IOP reduction.

Agreement between frequency doubling perimetry and static perimetry in eyes with high tension glaucoma and normal tension glaucoma.

AIMS: To investigate the agreement in results between frequency doubling technology (FDT) and the conventional automated static perimeter in eyes with normal tension glaucoma (NTG) and high tension glaucoma (HTG). METHODS: 72 eyes of 36 patients, who had two or more experiences with the Humphrey field analyser (HFA) program C30-2, were examined with the screening C-20-1 program of FDT. The result of FDT at each of the 17 stimulus points was graded as one of four categories. 58 out of 76 test points of HFA were assigned to one of the 17 clusters corresponding to FDT test points. Each cluster was represented as the lowest (scotoma of HFA) or the highest (threshold of HFA) probability symbol of total deviation (TD) of the HFA test points included in the cluster. The agreement between scotoma/threshold of HFA and FDT results was evaluated for NTG and HTG. RESULTS: In a total of 65 eyes, the Spearman coefficients between the FDT and HFA (threshold/scotoma of HFA) were 0.599 and 0.515 (p<0.0001), respectively. In the HFA mean deviation matched 20 HTG eyes and 20 NTG eyes, the number of points with abnormal FDT results were 102 and 62 in eyes with HTG and NTG, respectively. The eyes with HTG had more abnormal FDT results than NTG eyes (p=0.0014, Mann-Whitney U test). The kappa coefficient between FDT and threshold of HFA in eyes with HTG and NTG was 0.288 and 0.520, respectively, and the agreement between FDT and scotoma of HFA was 0.480 and 0.439, respectively. CONCLUSIONS: The best agreement of the results of FDT and HFA was observed in eyes with NTG using threshold of HFA. The eyes with HTG showed lower agreement with more abnormal points in FDT results, which suggests enough sensitivity of FDT in eyes with NTG, and higher sensitivity of FDT in eyes with HTG.

Acrolein produced from polyamines as one of the uraemic toxins.

It is well known that the addition of spermine or spermidine to culture medium containing ruminant serum inhibits cellular proliferation. This effect is caused by the products of oxidation of polyamines that are generated by serum amine oxidase. Among the products, we found that acrolein is a major toxic compound produced from spermine and spermidine by amine oxidase. We then analysed the level of polyamines (putrescine, spermidine and spermine) and amine oxidase activity in plasma of patients with chronic renal failure. It was found that the levels of putrescine and the amine oxidase activity were increased, whereas spermidine and spermine were decreased in plasma of patients with chronic renal failure. The levels of free and protein-conjugated acrolein were also increased in plasma of patients with chronic renal failure. An increase in putrescine, amine oxidase and acrolein in plasma was observed in all cases such as diabetic nephropathy, chronic glomerulonephritis and nephrosclerosis. These results suggest that acrolein is produced during the early stage of nephritis through kidney damage and also during uraemia through accumulation of polyamines in blood due to the decrease in their excretion into urine.

Effect of non-steroidal anti-inflammatory ophthalmic solution on intraocular pressure reduction by latanoprost.

AIM: To investigate the effects of a non-steroidal anti-inflammatory drug (NSAID) ophthalmic solution on latanoprost induced intraocular pressure (IOP) reduction using normal volunteers. METHODS: This study was conducted as a prospective and observer masked clinical trial. 13 normal volunteers were enrolled. After measurement of basal IOP and ophthalmic examination, latanoprost ophthalmic solution was initially administered to both eyes once daily. Four weeks later, an NSAID ophthalmic solution, sodium 2-amino-3-(4-bromobenzoyl) phenylacetate sesquihydrate (refer to bromfenac sodium hydrate), was co-administered to one randomly selected eye (NSAID group) twice daily for 2 weeks. The other eye was employed as a control (non-NSAID group). After withdrawal of the NSAID ophthalmic solution, latanoprost ophthalmic solution was continuously administered for another 2 weeks and was then withdrawn. After a 4 week washout, only bromfenac sodium hydrate ophthalmic solution was administered to the eyes of the NSAID group for 2 weeks. During the study period, ophthalmic examination, including IOP measurement was performed in an observer masked fashion. RESULTS: Before initiation of bromfenac sodium hydrate, baseline IOPs of the non-NSAID group and the NSAID group were 15.73 (SD 1.97) mm Hg and 15.86 (2.06) mm Hg, respectively (p=0.88). Although latanoprost ophthalmic solution significantly reduced IOP in both groups, co-administration of bromfenac sodium hydrate significantly inhibited latanoprost induced IOP reduction compared with the non-NSAID group. The IOPs of the non-NSAID and NSAID groups were 10.18 (1.17) mm Hg and 11.63 (1.35) mm Hg with a 2 week co-administration, respectively (p <0.01). Withdrawal of bromfenac sodium hydrate ophthalmic solution diminished the difference between the two groups. Re-administration of bromfenac sodium ophthalmic solution only did not affect IOP. CONCLUSION: These results indicate that NSAID ophthalmic solution may interfere with IOP reduction by latanoprost ophthalmic solution in normal volunteers and that we should take this into account when treating patients with glaucoma using latanoprost ophthalmic solution.

Iridial pigmentation induced by latanoprost ophthalmic solution in Japanese glaucoma patients.

PURPOSE: To investigate the incidence of iridial pigmentation induced by latanoprost ophthalmic solution in Japanese glaucoma patients by a prospective and observer-masked study. PATIENTS AND METHODS: Sixty-nine eyes of 69 glaucoma patients were included. Patients who had undergone intraocular surgery, laser trabeculoplasty, and laser iridotomy within 12 months before enrollment, and patients with history of uveitis and any changes in antiglaucoma drugs within 6 months before enrollment were excluded. Iridial photographs were taken by one examiner under the same conditions at 1, 3, and 6 months after the initiation of latanoprost treatment. Three glaucoma specialists, masked of patient information, independently assessed the iridial pigmentation. Cases with iridial pigmentation diagnosed by three specialists were categorized as showing a definite increase in iridial pigmentation. RESULTS: A definite increase in iridial pigmentation occurred in 3.5%, 9.7%, and 35.0% of eyes within 1, 3, and 6 months of treatment, respectively. Age, gender, or concomitantly used eyedrops did not significantly influence the incidence of iridial pigmentation within 6 months of instillation. A reduction of intraocular pressure by latanoprost did not differ significantly between patients with and without iridial pigmentation. CONCLUSION: The incidence of iridial pigmentation by latanoprost ophthalmic solution in Japanese patients was higher than previously reported values in pigmented races.

Effects of retinal glial cells on isolated rat retinal ganglion cells.

PURPOSE: The effect of retinal glial cells on retinal ganglion cell (RGC) survival was investigated in cocultures of pure, isolated retinal glial cells with pure, isolated RGCs. METHODS: RGCs from 2-day-old rats were cocultured for 48 hours, avoiding direct contact between cell types, with either nonconfluent retinal glial cells from 3-day-old rats or confluent retinal glial cells from 3-day-old, 12-day-old, or 1-year-old rats. Survival of RGCs was evaluated by flow cytometry. Amino acids were determined in culture medium. The effects of glutamate antagonists, 6-cyano-7-nitroquinoxaline-2,3-dione and MK801, a nitric oxide (NO) scavenger, 2-(4-carboxyphenyl)-4,4,5,5tetramethylimidazoline-1-oxyl-3-oxide potassium salt (c-PTIO), and an NO synthase inhibitor, N(G)-nitro-L-arginine methyl ester (L-NAME), were examined. RESULTS: Nonconfluent retinal glial cells significantly reduced the survival of small and large RGCs, but confluent retinal glial cells reduced the survival of only small RGCs, regardless of the rat's age at the time of retinal glial cell harvesting. Profiles of some amino acids significantly varied, depending on the culture condition. Cocultures of RGCs with nonconfluent retinal glial cells released significantly more glutamate into the medium than cocultures of RGCs with confluent retinal glial cells or RGCs in pure culture. The glutamate antagonists improved the survival of RGCs cocultured with nonconfluent retinal glial cells, especially when the two were administered in combination, and in the case of large RGCs. c-PTIO and L-NAME, also improved the survival of RGCs cocultured with nonconfluent retinal glial cells. CONCLUSIONS: Adverse effects of retinal glial cells on the survival of RGCs varied by size of the RGCs and retinal glial cell confluence. Glutamate and NO may be involved in retinal glial cell-related antisurvival effects.