Neuroprotective effects of bis(7)-tacrine against glutamate-induced retinal ganglion cells damage
© Fang et al; licensee BioMed Central Ltd. 2010
Received: 23 October 2009
Accepted: 3 March 2010
Published: 3 March 2010
Glutamate-mediated excitotoxicity, primarily through N-methyl-D-aspartate (NMDA) receptors, may be an important cause of retinal ganglion cells (RGCs) death in glaucoma and several other retinal diseases. Bis(7)-tacrine is a noncompetitive NMDA receptors antagonist that can prevent glutamate-induced hippocampal neurons damage. We tested the effects of bis(7)-tacrine against glutamate-induced rat RGCs damage in vitro and in vivo.
In cultured neonatal rats RGCs, the MTT assay showed that glutamate induced a concentration- and time-dependent toxicity. Bis(7)-tacrine and memantine prevented glutamate-induced cell death in a concentration-dependent manner with IC50 values of 0.028 μM and 0.834 μM, respectively. The anti-apoptosis effects of bis(7)-tacrine were confirmed by annexin V-FITC/PI staining. In vivo, TUNEL analysis and retrograde labeling analysis found that pretreatment with bis(7)-tacrine(0.2 mg/kg) induced a significant neuroprotective effect against glutamate-induced RGCs damage.
Our results showed that bis(7)-tacrine had neuroprotective effects against glutamate-induced RGCs damage in vitro and in vivo, possibly through the drug's anti-NMDA receptor effects. These findings make bis(7)-tacrine potentially useful for treating a variety of ischemic or traumatic retinopathies inclusive of glaucoma.
Glutamate is a major excitatory neurotransmitter in the central nervous system, including the retina[1, 2]. It is released by the presynaptic cells and acts on N-methyl-D-aspartate (NMDA), α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA), and kainite (KA) receptors . If excessive amounts of glutamate are released or if glutamate clearance is insufficient, neuronal death will result in a process known as excitotoxicity. The glutamate receptor-mediated excitotoxicity has been associated to various diseases of the brain and eye, which include Alzheimer's disease, retinal ischemia[5, 6] and glaucoma[7, 8]. Although retinal ganglion cells (RGCs) express all of three receptor subtypes, the glutamate toxicity is primarily mediated by NMDA receptors [9–11].
Glaucoma, a neurodegenerative disease, is associated with selective death of retinal ganglion cells . The disease is characterized by an elevation in intraocular pressure (IOP), which leads to increased glutamate levels . Vitreal glutamate levels are elevated in dogs and humans with primary glaucoma , and also in monkeys with experimentally induced chronic glaucoma. Lowering IOP is the current main treatment for glaucoma, yet disease progression continues to occur even in patients with significant IOP reduction. Therefore lowering IOP is inadequate for glaucoma patients [12, 18]. Efforts have been made to attempt to discover appropriate drugs or chemicals (neuroprotectants) that can be taken orally to slow down retinal ganglion cell death and have negligible side-effects . Memantine is an uncompetitive NMDA receptor antagonist which is prescribed for the treatment of Alzheimer's disease . However, two recent parallel clinical trials conducted to test the efficacy of memantine as a neuroprotectant for glaucoma were unsuccessful. The results of the trials showed that memantine had no significant effects in preserving visual function.
Until now, there has been no neuroprotective agent indicated for the treatment of glaucoma. A neuroprotectant that has a single mode of action like memantine has a limited positive effect in slowing down ganglion cell death. Pharmacological agents that simultaneously affect multiple biological mechanisms are consequently desired. This has been referred to as the "cocktail" approach . One-drug-multiple-targets approach in the treatment of neurodegenerative diseases is possible way forward [19, 23].
Beside NMDA receptor antagonism, other strategies have been investigated in the development of neuroprotective therapies, which include voltage-dependent calcium channel blockade [24, 25], nitric oxide synthase (NOS) inhibition [26, 27], and so on. Bis(7)-tacrine (1,7-N-heptylene-bis-9,9'-amino-1,2,3,4-tetrahydroacridine), a dimeric acetylcholinesterase (AChE) inhibitor derived from anti-Alzheimer's drug tacrine, possesses remarkable neuroprotective activities through concurrent inhibition of AChE [28, 29], NMDA receptor and nitric oxide synthase [31, 32]. Moreover, bis(7)-tacrine attenuates neuronal apoptosis by regulating L-type voltage-dependent calcium channels(VDCCs) . Recent studies showed that bis(7)-tacrine prevented glutamate-induced excitotoxicity by selectively inhibiting NMDA receptors in primary cultured cerebellar granule neurons (CGNs), without the involvement of the other two ionotropic glutamate receptors, AMPA receptor and KA receptor [30, 34, 35].
Based on these evidence, we hypothesis that bis(7)-tacrine attenuate glutamate-induced retinal ganglion cells damage through the blockade of NMDA receptors. We tested the effect of bis(7)-tacrine in two models of glutamate excitotoxity, RGCs in culture with glutamate and intravitreal injection of glutamate. The results showed that bis(7)-tacrine reduced glutamate-induced retinal ganglion cells damage in vitro and in vivo.
Identification of cultured RGCs
Bis(7)-tacrine prevents glutamate-induced cell death more potently than memantine
Bis(7)-tacrine reduces glutamate-induced apoptosis of RGCs in vitro
Bis(7)-tacrine reduces glutamate-induced apoptosis of RGCs in vivo
Bis(7)-tacrine protect RGCs in experiments using retrograde labeling of RGCs
Glutamate excitotoxicity is thought to contribute to a broad variety of neurological diseases, including Alzheimer's disease and glaucoma. In retina tissues, the predominant form of glutamate neurotoxicity is mediated by overstimulation of the NMDA subtype of glutamate receptors, which in turn causes an increase of Ca2+ influx, followed by cell death [5, 9, 38]. Recent research demonstrated that bis(7)-tacrine prevented glutamate-induced cerebellar granule neurons (CGNs) apoptosis through directly blocking NMDA receptors [30, 34]. We show here that bis(7)-tacrine provided in vitro and in vivo neuroprotective effects on glutamate-induced RGCs damage. Although the cytoprotective actions of bis(7)-tacrine have previously been observed in brain neurons under excitotoxic/ischemic conditions[23, 30, 34, 39], to our knowledge, this is the first report to elucidate the neuroprotective action of bis(7)-tacrine on RGCs in response to glutamate excitotoxicity.
Some researches have reported that RGCs are highly susceptible to glutamate toxicity in vitro and in vivo[9, 38, 40]. First of all, we explored the in vitro effectiveness of bis(7)-tacrine against glutamate-induced RGCs damage. In a previous report, glutamate toxicity was concentration-dependent with a calculated EC50 of 30.8 μM. In our study, the MTT assays have shown that cultured RGCs are highly sensitive to 24 h 50 μM glutamate treatment, which produces 50% or greater cell death. Furthermore, bis(7)-tacrine, like the NMDA receptor antagonist memantine, has been confirmed to attenuate the cytotoxic effect of glutamate. As memantine showed affinity and potency comparable to those of bis(7)-tacrine in blocking the NMDA receptor , we chose memantine as the positive control in this study. The minimal effective concentration of bis(7)-tacrine against glutamate excitotoxicity was approximately 0.01 μM. The combination of data from the MTT assays and the Annexin V-FITC/PI assays, provided strong evidence that bis(7)-tacrine was significantly more potent than memantine in inhibiting glutamate-induced cell damage. This finding was similar to previous reports in cultured rat cortical neurons.
Next, we verified the neuroprotective effects of bis(7)-tacrine in animal models of glutamate-induced retinal injury. The pharmacokinetic research in the body of the rat showed that bis(7)-tacrine was rapidly and widely distributed to its target tissues such as brain[41, 42]. In a model of acute focal cerebral ischemic insults in MCAO rats, bis(7)-tacrine at doses of 0.1 mg/kg and 0.2 mg/kg significantly reduced ischemic impairment in vivo. In the current study, two animal assays showed that bis(7)-tacrine(0.2 mg/kg) protected RGCs from glutamate excitotoxicity, while bis(7)-tacrine(0.05 mg/kg) had no significant neuroprotective effects.
We cannot definitively state that bis(7)-tacrine was neuroprotective for retinal ganglion cells alone. In retina tissues, several cells express NMDA receptors, such as RGCs and displaced amacrine cells in the retinal ganglion cell layer (GCL)and bipolar cells in the inner nuclear layer (INL). These cells are susceptible to excitotoxic cell death, we did not differentiate between retinal ganglion cells and other cells.
Neuroprotection was initially studied as a treatment strategy for various neurological disorders including stroke, dementia, multiple sclerosis, Alzheimer's disease and glaucoma. Despite successful preclinical cell culture and animal model experiments, most of these therapies were not successful at the clinical stage of testing because of either unacceptable side effect profiles or a lack of efficacy [12, 45, 46]. High affinity NMDA receptor antagonists such as MK-801 are undesirable as they entail nonselective inhibition of tonic glutamate activity as well as phasic physiological NMDA receptor function . Memantine is a moderate affinity, uncompetitive NMDA receptor antagonist prescribed for the treatment of moderate to severe Alzheimer's disease [48, 49]. However, recent clinical findings showed that there was no clear benefit after glaucoma patients received memantine; thereby it was suggested that neuroprotectants with multiple modes of actions were likely to reveal clearer results than was found for memantine .
The preclinical studies demonstrated that bis(7)-tacrine had low toxicity in animal models, and bis(7)-tacrine possessed multiple physiological activities including anti-NMDA receptors, anti-AChE, anti-L-type-voltage-dependent calcium channels(VDCCs), anti-nitric oxide synthase (NOS) signaling and the regulation of the downstream signal of NMDA receptors [28, 30, 32, 33, 51]. In the present study, bis(7)-tacrine prevented glutamate-induced RGCs damage possibly by inhibiting NMDA receptors. At this point, electrophysiology studies will be needed to verify the blocking kinetics of bis(7)-tacrine on three glutamate receptors in the purified RGCs. What's more, further calcium imaging studies will be required to determine the role of calcium permeation through NMDA receptors and to determine if intracellular calcium is involved in triggering neuroprotection or inhibiting glutamate excitotoxicity. Besides inhibiting NMDA receptors, the other effects of bis(7)-tacrine on RGCs deserves further study.
In conclusion, our experiments have demonstrated that bis(7)-tacrine can provide neuroprotection against glutamate-induced retinal ganglion cells damage in vitro and in vivo. The effects may be achieved through inhibitions of NMDA receptors. This neuroprotective effects of bis(7)-tacrine may lead to a novel approach for the treatment of retinopathies, such as glaucoma.
Animals and reagents
Sprague-Dawley(SD) rats, including 1-3 days rats and adult male rats, were obtained from the Animal Center in Tongji Medical College, Huazhong University of Science and Technology, and were housed in the animal facility under standard conditions of room temperature and a 12:12 h light-dark cycle with free access to food and water. All animal experiments followed the guidelines for the care and use of animals established by Tongji Medical College, Huazhong University of Science and Technology and adhered to the tenets of the Declaration of Helsinki. Bis(7)-tacrine was purchased from Cayman Chemical Co.(USA). Fluorogold was purchased from Biotium (Hayward, USA). Unless noted, all other reagents were obtained from Sigma (St. Louis, MO, USA).
Cell culture and purification of RGCs
RGCs from retinas of 1-3 days rats were purified by a two-step immunopanning procedure. Briefly, the retinal tissue was dissociated into single cells in EMEM (Gibco, China) containing 15 U/ml papain and 70 U/ml collagenase. The dissociated cells were incubated in a polypropylene tube coated with an anti-rat macrophage monoclonal IgG (Chemicon International, Inc, CA, USA) to exclude macrophages, and then incubated in a tube coated with an anti-rat Thy 1.1 monoclonal IgG (Chemicon International, Inc, CA, USA). The tube was gently washed with PBS for five times, and adherent RGCs were collected by centrifugation at 600 g for 5 minutes.
Before the examinations of effects of bis(7)-tacrine on RGCs, a preliminary study was conducted to determine purity of RGCs after the two-step immunopanning procedure. After 1-day-old rats were anesthetized by intraperitoneal injection with 10% (w/v) chloral hydrate (350 mg/kg), RGCs were labeled in a retrograde manner by injecting 4% fluorogold into the superior colliculi. Four days later, after this immunopanning method, approximately 87.8% (1896/2158 cells) of the collected cells were labeled by fluorogold. Next, in further examinations of effects of bis(7)-tacrine on RGCs, RGCs were used from rats without fluorogold injection and grown in serum-free medium (Gibco-China), containing 1 mM glutamine, 10 μg/mL gentamicin, B27 supplement (1:50), 40 ng/mL each of BDNF and CNTF, and 5 μM forskolin. RGCs were cultured in a CO2 incubator (Thermo Lab 2300, USA) with 5% CO2 at 37°C. Before seeding, the plates were coated with poly-D-lysine (PDL, 70 kDa, 10 μg/ml) at room temperature followed by overnight incubation with mouse laminin. RGCs were cultured for 3 days and then exposed them to glutamate and/or bis(7)-tacrine for MTT assay or annexin-V FITC/PI assay.
Measurement of neurotoxicity
The percentage of surviving RGCs in the presence of bis(7)-tacrine and/or glutamate was estimated by determining the activity of mitochondrial dehydrogenases with 3(4,5-dimethylthiazol-2-yl)-2.5-diphenyltetrazolium bromide (MTT) assay . Cells were seeded at a density of 5000 cells/well in 96-well plates. The assay was performed according to the specifications of the manufacturer (MTT kit I; Roche China, Ltd.). Briefly, the rat RGCs were cultured in 96-well plates, 10 μl of 5 mg/ml MTT labeling reagent was added to each well containing cells in 100 μl of medium, and the plate was incubated for 4 h in a humidified incubator at 37°C. After the incubation, 100 μl of the solvating solution (0.01 N HCl in 10% SDS solution) was added to each well for 17-18 h. The absorbance of the samples was measured at a wavelength of 570 nm with 630 nm as a reference wavelength. Unless otherwise indicated, the extent of MTT conversion in cells exposed to glutamate is expressed as a percentage of the control.
Annexin-V FITC/PI assay
Apoptosis was detected using an Annexin-V FITC/PI detection kit (Jiancheng Biotechnology Co., Ltd., Nanjing, China) according to the manufacturer's directions. The cells were digested with 0.125% trypsin, washed with ice-cold phosphate-buffered saline and resuspended in binding buffer (5 × 105 cells/ml). Then, the cells were centrifuged at 1,000 rpm for 5 min at 4°C. After the supernatant had been discarded, 500 μl of binding buffer, 5 μl of annexin-V-FITC and 5 μl of propidium iodide (PI) were added to 200 μl of the cell suspension. After mixing gently, the suspensions were incubated for 15 min at room temperature without light. Finally, the cells were analyzed by flow cytometry (BD LSRII; BD Biosciences).
Intravitreal administration of glutamate
Male rats (220-280 g) were anesthetized by intraperitoneal injection of 10% (w/v) chloral hydrate (350 mg/kg) and rectal body temperature was maintained at 37°C with a heating pad during the experiments. The pupils were dilated with tropicamide and a single dose of 5 μl of 4 mM glutamate (total amount 20 nmol) in 0.01 M PBS (pH 7.4) was injected into the vitreous cavity using 32-gauge Hamilton needle and syringe. PBS was administered as a control.
TUNEL staining was performed according to the manufacturer's protocols (In Situ Cell Death Detection Kit; Roche China, Ltd.) to detect retinal cell apoptosis induced by glutamate. Twenty-four rats were divided into eight groups: three bis(7)-tacrine-treated groups (0.05, 0.1, 0.2 mg/kg), three memantine-treated groups (5, 10, 20 mg/kg), a control group, and a glutamate group. Six eyes per experimental condition were used. In this study, 18 h after the glutamate injection, rats were killed with an overdose of chloral hydrate. The eyes were immediately enucleated and fixed in 4% paraformaldehyde in PBS for the TUNEL studies. The specimens were then dehydrated and embedded in paraffin and 5 μm sections were cut. These sections were stained by the TUNEL method according to the manufacturer's directions. The yellow condensed TUNEL positive cells were counted under a 20× objective microscope. No attempt was made to distinguish the cell types in the GCL, and displaced amacrine cells were not excluded from the counts. To minimize the variance in cell number, we counted TUNEL positive cells in the retinal ganglion cell layer (GCL) manually at 1.0-2.0 mm (both sides) from the center of the optic disc. The average number of TUNEL positive cells/eye was obtained from three sections of each retina.
Retrograde labelling and counting of RGCs
To examine the change in the number of RGCs after glutamate injection, RGCs were retrogradely labeled with fluorogold. Twenty-four rats were treated respectively with the same division and administration above mentioned. Fifteen minutes before intravitreally injection of glutamate, drugs were intraperitoneally administered to the rats in a volume of 1.5 mL/kg body weight. Four days after the glutamate injection, retrograde labeling of RGCs was made. Briefly, rats were anesthetized by intraperitoneally injection with 10% (w/v) chloral hydrate (350 mg/kg) and then the heads were fixed in a stereotaxic apparatus. Fluorogold was microinjected bilaterally into the superior colliculi(SC) and dorsal lateral geniculate nuclei (dLGN) of rats. Three days after the Fluorogold injection (seven days after the glutamate injection), the animals were killed by an intraperitoneal overdose injection of chloral hydrate and the eyes were enucleated. Eyes were fixed with 4% paraformaldehyde for 1 h. Retinas were removed from the sclera, divided into four radial cuts and mounted on slides. Analysis for the number of fluorogold-labeled RGCs was carried out. Briefly, Tracer-labeled RGCs counting was performed in 12 areas of 0.072 mm2 each (three areas per retinal quadrant) at 2/6, 3/6, and 5/6 of the retinal radius under a fluorescent microscope (Olympus IX71). Data from 12 areas from each eye were averaged.
where X and Y are concentration and response, respectively, Emax is the maximal response, EC50 is the concentration yielding 50% of maximal effect (EC50 for activation, IC50 for inhibition), and n is the slope factor.
- Lucas DR, Newhouse JP: The toxic effect of sodium L-glutamate on the inner layers of the retina. AMA Arch Ophthalmol. 1957, 58 (2): 193-201.View ArticlePubMedGoogle Scholar
- Thoreson WB, Witkovsky P: Glutamate receptors and circuits in the vertebrate retina. Prog Retin Eye Res. 1999, 18 (6): 765-810. 10.1016/S1350-9462(98)00031-7.View ArticlePubMedGoogle Scholar
- Yu W, Miller RF: The mechanism by which NBQX enhances NMDA currents in retinal ganglion cells. Brain Res. 1996, 709 (2): 184-196. 10.1016/0006-8993(95)01285-0.View ArticlePubMedGoogle Scholar
- Sonkusare SK, Kaul CL, Ramarao P: Dementia of Alzheimer's disease and other neurodegenerative disorders--memantine, a new hope. Pharmacol Res. 2005, 51 (1): 1-17.View ArticlePubMedGoogle Scholar
- Sucher NJ, Lipton SA, Dreyer EB: Molecular basis of glutamate toxicity in retinal ganglion cells. Vision Res. 1997, 37 (24): 3483-3493. 10.1016/S0042-6989(97)00047-3.View ArticlePubMedGoogle Scholar
- Osborne NN, Casson RJ, Wood JP, Chidlow G, Graham M, Melena J: Retinal ischemia: mechanisms of damage and potential therapeutic strategies. Prog Retin Eye Res. 2004, 23 (1): 91-147. 10.1016/j.preteyeres.2003.12.001.View ArticlePubMedGoogle Scholar
- Dreyer EB: A proposed role for excitotoxicity in glaucoma. J Glaucoma. 1998, 7 (1): 62-67. 10.1097/00061198-199802000-00012.View ArticlePubMedGoogle Scholar
- Kuehn MH, Fingert JH, Kwon YH: Retinal ganglion cell death in glaucoma: mechanisms and neuroprotective strategies. Ophthalmol Clin North Am. 2005, 18 (3): 383-395. 10.1016/j.ohc.2005.04.002. viView ArticlePubMedGoogle Scholar
- Pang IH, Zeng H, Fleenor DL, Clark AF: Pigment epithelium-derived factor protects retinal ganglion cells. BMC Neurosci. 2007, 8: 11-10.1186/1471-2202-8-11.PubMed CentralView ArticlePubMedGoogle Scholar
- Inomata Y, Hirata A, Yonemura N, Koga T, Kido N, Tanihara H: Neuroprotective effects of interleukin-6 on NMDA-induced rat retinal damage. Biochem Biophys Res Commun. 2003, 302 (2): 226-232. 10.1016/S0006-291X(03)00127-X.View ArticlePubMedGoogle Scholar
- Calzada JI, Jones BE, Netland PA, Johnson DA: Glutamate-induced excitotoxicity in retina: neuroprotection with receptor antagonist, dextromethorphan, but not with calcium channel blockers. Neurochem Res. 2002, 27 (1-2): 79-88. 10.1023/A:1014854606309.View ArticlePubMedGoogle Scholar
- Levin LA, Peeples P: History of neuroprotection and rationale as a therapy for glaucoma. Am J Manag Care. 2008, 14 (1 Suppl): S11-14.PubMedGoogle Scholar
- Lipton SA: Possible role for memantine in protecting retinal ganglion cells from glaucomatous damage. Surv Ophthalmol. 2003, 48 (Suppl 1): S38-46. 10.1016/S0039-6257(03)00008-0.View ArticlePubMedGoogle Scholar
- Fatma N, Kubo E, Sen M, Agarwal N, Thoreson WB, Camras CB, Singh DP: Peroxiredoxin 6 delivery attenuates TNF-alpha-and glutamate-induced retinal ganglion cell death by limiting ROS levels and maintaining Ca(2+) homeostasis. Brain Res. 2008, 3;1233: 63-78. 10.1016/j.brainres.2008.07.076.View ArticleGoogle Scholar
- Brooks DE, Garcia GA, Dreyer EB, Zurakowski D, Franco-Bourland RE: Vitreous body glutamate concentration in dogs with glaucoma. Am J Vet Res. 1997, 58 (8): 864-867.PubMedGoogle Scholar
- Dreyer EB, Zurakowski D, Schumer RA, Podos SM, Lipton SA: Elevated glutamate levels in the vitreous body of humans and monkeys with glaucoma. Arch Ophthalmol. 1996, 114 (3): 299-305.View ArticlePubMedGoogle Scholar
- Levin LA: Retinal ganglion cells and neuroprotection for glaucoma. Surv Ophthalmol. 2003, 48 (Suppl 1): S21-24. 10.1016/S0039-6257(03)00007-9.View ArticlePubMedGoogle Scholar
- Nagata T, Ueno S, Morita H, Kubota T, Toyohira Y, Tsutsui M, Tawara A, Yanagihara N: Direct inhibition of N-methyl-D-aspartate (NMDA)-receptor function by antiglaucomatous beta-antagonists. J Pharmacol Sci. 2008, 106 (3): 423-434. 10.1254/jphs.FP0071776.View ArticlePubMedGoogle Scholar
- Osborne NN: Recent clinical findings with memantine should not mean that the idea of neuroprotection in glaucoma is abandoned. Acta Ophthalmol. 2009, 87 (4): 450-454. 10.1111/j.1755-3768.2008.01459.x.View ArticlePubMedGoogle Scholar
- Molinaro G, Battaglia G, Riozzi B, Di Menna L, Rampello L, Bruno V, Nicoletti F: Memantine treatment reduces the expression of the K(+)/Cl(-) cotransporter KCC2 in the hippocampus and cerebral cortex, and attenuates behavioural responses mediated by GABA(A) receptor activation in mice. Brain Res. 2009, 1265: 75-79. 10.1016/j.brainres.2009.02.016.View ArticlePubMedGoogle Scholar
- Danesh-Meyer HV, Levin LA: Neuroprotection: extrapolating from neurologic diseases to the eye. Am J Ophthalmol. 2009, 148 (2): 186-191. 10.1016/j.ajo.2009.03.029.View ArticlePubMedGoogle Scholar
- Recommendations for standards regarding preclinical neuroprotective and restorative drug development. Stroke. 1999, 30 (12): 2752-2758.Google Scholar
- Zhao Y, Li W, Chow PC, Lau DT, Lee NT, Pang Y, Zhang X, Wang X, Han Y: Bis(7)-tacrine, a promising anti-Alzheimer's dimer, affords dose- and time-dependent neuroprotection against transient focal cerebral ischemia. Neurosci Lett. 2008, 439 (2): 160-164. 10.1016/j.neulet.2008.05.007.View ArticlePubMedGoogle Scholar
- Netland PA, Chaturvedi N, Dreyer EB: Calcium channel blockers in the management of low-tension and open-angle glaucoma. Am J Ophthalmol. 1993, 115 (5): 608-613.View ArticlePubMedGoogle Scholar
- Kanellopoulos AJ, Erickson KA, Netland PA: Systemic calcium channel blockers and glaucoma. J Glaucoma. 1996, 5 (5): 357-362. 10.1097/00061198-199610000-00011.View ArticlePubMedGoogle Scholar
- Neufeld AH, Sawada A, Becker B: Inhibition of nitric-oxide synthase 2 by aminoguanidine provides neuroprotection of retinal ganglion cells in a rat model of chronic glaucoma. Proc Natl Acad Sci USA. 1999, 96 (17): 9944-9948. 10.1073/pnas.96.17.9944.PubMed CentralView ArticlePubMedGoogle Scholar
- Neufeld AH: Pharmacologic neuroprotection with an inhibitor of nitric oxide synthase for the treatment of glaucoma. Brain Res Bull. 2004, 62 (6): 455-459. 10.1016/j.brainresbull.2003.07.005.View ArticlePubMedGoogle Scholar
- Pang YP, Quiram P, Jelacic T, Hong F, Brimijoin S: Highly potent, selective, and low cost bis-tetrahydroaminacrine inhibitors of acetylcholinesterase. Steps toward novel drugs for treating Alzheimer's disease. J Biol Chem. 1996, 271 (39): 23646-23649. 10.1074/jbc.271.39.23646.View ArticlePubMedGoogle Scholar
- Wang H, Carlier PR, Ho WL, Wu DC, Lee NT, Li CP, Pang YP, Han YF: Effects of bis(7)-tacrine, a novel anti-Alzheimer's agent, on rat brain AChE. Neuroreport. 1999, 10 (4): 789-793.View ArticlePubMedGoogle Scholar
- Li W, Pi R, Chan HH, Fu H, Lee NT, Tsang HW, Pu Y, Chang DC, Li C, Luo J, et al.: Novel dimeric acetylcholinesterase inhibitor bis7-tacrine, but not donepezil, prevents glutamate-induced neuronal apoptosis by blocking N-methyl-D-aspartate receptors. J Biol Chem. 2005, 280 (18): 18179-18188. 10.1074/jbc.M411085200.View ArticlePubMedGoogle Scholar
- Li W, Mak M, Jiang H, Wang Q, Pang Y, Chen K, Han Y: Novel Anti-Alzheimer's Dimer Bis(7)-Cognitin: Cellular and Molecular Mechanisms of Neuroprotection Through Multiple Targets. Neurotherapeutics. 2009, 6 (1): 187-201. 10.1016/j.nurt.2008.10.040.View ArticlePubMedGoogle Scholar
- Li W, Lee NT, Fu H, Kan KK, Pang Y, Li M, Tsim KW, Li X, Han Y: Neuroprotection via inhibition of nitric oxide synthase by bis(7)-tacrine. Neuroreport. 2006, 17 (5): 471-474. 10.1097/01.wnr.0000209014.09094.72.View ArticlePubMedGoogle Scholar
- Fu H, Li W, Lao Y, Luo J, Lee NT, Kan KK, Tsang HW, Tsim KW, Pang Y, Li Z, et al.: Bis(7)-tacrine attenuates beta amyloid-induced neuronal apoptosis by regulating L-type calcium channels. J Neurochem. 2006, 98 (5): 1400-1410. 10.1111/j.1471-4159.2006.03960.x.View ArticlePubMedGoogle Scholar
- Liu YW, Li CY, Luo JL, Li WM, Fu HJ, Lao YZ, Liu LJ, Pang YP, Chang DC, Li ZW, et al.: Bis(7)-tacrine prevents glutamate-induced excitotoxicity more potently than memantine by selectively inhibiting NMDA receptors. Biochem Biophys Res Commun. 2008, 369 (4): 1007-1011. 10.1016/j.bbrc.2008.02.133.View ArticlePubMedGoogle Scholar
- Luo J, Li W, Liu Y, Zhang W, Fu H, Lee NT, Yu H, Pang Y, Huang P, Xia J, et al.: Novel dimeric bis(7)-tacrine proton-dependently inhibits NMDA-activated currents. Biochem Biophys Res Commun. 2007, 361 (2): 505-509. 10.1016/j.bbrc.2007.07.043.View ArticlePubMedGoogle Scholar
- Otori Y, Wei JY, Barnstable CJ: Neurotoxic effects of low doses of glutamate on purified rat retinal ganglion cells. Invest Ophthalmol Vis Sci. 1998, 39 (6): 972-981.PubMedGoogle Scholar
- Linden R, Esberard CE: Displaced amacrine cells in the ganglion cell layer of the hamster retina. Vision Res. 1987, 27 (7): 1071-1076. 10.1016/0042-6989(87)90021-6.View ArticlePubMedGoogle Scholar
- Kawasaki A, Han MH, Wei JY, Hirata K, Otori Y, Barnstable CJ: Protective effect of arachidonic acid on glutamate neurotoxicity in rat retinal ganglion cells. Invest Ophthalmol Vis Sci. 2002, 43 (6): 1835-1842.PubMedGoogle Scholar
- Li W, Xue J, Niu C, Fu H, Lam CS, Luo J, Chan HH, Xue H, Kan KK, Lee NT, et al.: Synergistic neuroprotection by bis(7)-tacrine via concurrent blockade of N-methyl-D-aspartate receptors and neuronal nitric-oxide synthase. Mol Pharmacol. 2007, 71 (5): 1258-1267. 10.1124/mol.106.029108.View ArticlePubMedGoogle Scholar
- Kawasaki A, Otori Y, Barnstable CJ: Muller cell protection of rat retinal ganglion cells from glutamate and nitric oxide neurotoxicity. Invest Ophthalmol Vis Sci. 2000, 41 (11): 3444-3450.PubMedGoogle Scholar
- Li Z, Hua Y, Ming LW, Chun CM, Ping PY, Ge L, Tao WY, Zhong Z, Fan HY: Selective and sensitive determination of bis(7)-tacrine, a high erythrocyte binding acetylcholinesterase inhibitor, in rat plasma by high-performance liquid chromatography-tandem mass spectrometry. Biomed Chromatogr. 2008, 22 (4): 414-420. 10.1002/bmc.949.View ArticlePubMedGoogle Scholar
- Yu H, Ho JM, Kan KK, Cheng BW, Li WM, Zhang L, Lin G, Pang YP, Gu ZM, Chan K, et al.: Development of a high performance liquid chromatography-tandem mass method for determination of bis(7)-tacrine, a promising anti-Alzheimer's dimer, in rat blood. J Pharm Biomed Anal. 2007, 44 (5): 1133-1138. 10.1016/j.jpba.2007.05.028.View ArticlePubMedGoogle Scholar
- Brandstatter JH, Hartveit E, Sassoe-Pognetto M, Wassle H: Expression of NMDA and high-affinity kainate receptor subunit mRNAs in the adult rat retina. Eur J Neurosci. 1994, 6 (7): 1100-1112. 10.1111/j.1460-9568.1994.tb00607.x.View ArticlePubMedGoogle Scholar
- Costenla AR, De Mendonca A, Sebastiao A, Ribeiro JA: An adenosine analogue inhibits NMDA receptor-mediated responses in bipolar cells of the rat retina. Exp Eye Res. 1999, 68 (3): 367-370. 10.1006/exer.1998.0645.View ArticlePubMedGoogle Scholar
- Muir KW, Grosset DG: Neuroprotection for acute stroke: making clinical trials work. Stroke. 1999, 30 (1): 180-182.View ArticlePubMedGoogle Scholar
- Hickenbottom SL, Grotta J: Neuroprotective therapy. Semin Neurol. 1998, 18 (4): 485-492. 10.1055/s-2008-1040901.View ArticlePubMedGoogle Scholar
- Volbracht C, van Beek J, Zhu C, Blomgren K, Leist M: Neuroprotective properties of memantine in different in vitro and in vivo models of excitotoxicity. Eur J Neurosci. 2006, 23 (10): 2611-2622. 10.1111/j.1460-9568.2006.04787.x.View ArticlePubMedGoogle Scholar
- Gilling KE, Jatzke C, Hechenberger M, Parsons CG: Potency, voltage-dependency, agonist concentration-dependency, blocking kinetics and partial untrapping of the uncompetitive N-methyl-D-aspartate (NMDA) channel blocker memantine at human NMDA (GluN1/GluN2A) receptors. Neuropharmacology. 2009, 56 (5): 866-875. 10.1016/j.neuropharm.2009.01.012.View ArticlePubMedGoogle Scholar
- Chen HS, Lipton SA: Mechanism of memantine block of NMDA-activated channels in rat retinal ganglion cells: uncompetitive antagonism. J Physiol. 1997, 499 (Pt 1): 27-46.PubMed CentralView ArticlePubMedGoogle Scholar
- Liu J, Ho W, Lee NT, Carlier PR, Pang Y, Han Y: Bis(7)-tacrine, a novel acetylcholinesterase inhibitor, reverses AF64A-induced deficits in navigational memory in rats. Neurosci Lett. 2000, 282 (3): 165-168. 10.1016/S0304-3940(00)00905-8.View ArticlePubMedGoogle Scholar
- Liu YW, Luo JL, Ren H, Peoples RW, Ai YX, Liu LJ, Pang YP, Li ZW, Han YF, Li CY: Inhibition of NMDA-gated ion channels by bis(7)-tacrine: whole-cell and single-channel studies. Neuropharmacology. 2008, 54 (7): 1086-1094. 10.1016/j.neuropharm.2008.02.015.View ArticlePubMedGoogle Scholar
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