Pathophysiology of human glaucomatous optic nerve damage: Insights from rodent models of glaucoma

Understanding mechanisms of glaucomatous optic nerve damage is essential for developing effective therapies to augment conventional pressure-lowering treatments. This requires that we understand not only the physical forces in play, but the cellular responses that translate these forces into axonal injury. The former are best understood by using primate models, in which a well-developed lamina cribrosa, peripapillary sclera and blood supply are most like that of the human optic nerve head. However, determining cellular responses to elevated intraocular pressure (IOP) and relating their contribution to axonal injury require cell biology techniques, using animals in numbers sufficient to perform reliable statistical analyses and draw meaningful conclusions. Over the years, models of chronically elevated IOP in laboratory rats and mice have proven increasingly useful for these purposes. While lacking a distinct collagenous lamina cribrosa, the rodent optic nerve head (ONH) possesses a cellular arrangement of astrocytes, or glial lamina, that ultrastructurally closely resembles that of the primate. Using these tools, major insights have been gained into ONH and the retinal cellular responses to elevated IOP that, in time, can be applied to the primate model and, ultimately, human glaucoma.     

The optic nerve head in glaucoma: role of astrocytes in tissue remodeling

Primary open angle glaucoma is a common eye disease characterized by loss of the axons of the retinal ganglion cells leading to progressive loss of vision. The site of damage to the axons is at the level of the lamina cribrosa in the optic nerve head. The mechanism of axonal loss is unknown but elevated intraocular pressure and age are the most common factors associated with the disease. Previous studies in human glaucoma and in experimental glaucoma in monkeys have established a relationship between chronic elevation of intraocular pressure and remodeling of the optic nerve head tissues known clinically as cupping of the optic disc. This review focuses on the astrocytes, the major cell type in the optic nerve head. Astrocytes participate actively in the remodeling of neural tissues during development and in disease. In glaucomatous optic neuropathy, astrocytes play a major role in the remodeling of the extracellular matrix of the optic nerve head, synthesize growth factors and other cellular mediators that may affect directly, or indirectly, the axons of the retinal ganglion cells. Due to the architecture of the lamina cribrosa, formed by the cells and the fibroelastic extracellular matrix, astrocytes may respond to changes in intraocular pressure in glaucoma, leading to some of the detrimental events that underlie axonal loss and retinal ganglion cell degeneration.     

Understanding mechanisms of pressure-induced optic nerve damage

Patients with glaucoma can suffer progressive vision loss, even in the face of what appears to be excellent intraocular pressure (IOP) control. Some of this may be secondary to non-pressure-related (pressure-independent) factors. However, it is likely that chronically elevated IOP produces progressive changes in the optic nerve head, the retina, or both that alter susceptibility of remaining optic nerve fibers to IOP. In order to understand the nature of these progressive changes, relevant, cost-effective animal models are necessary. Several rat models are now used to produce chronic, elevated IOP, and methods exist for measuring the resulting IOP and determining the extent of the damage this causes to the retina and optic nerve. A comparison of damage, pressure and duration shows that these models are not necessarily equivalent. These tools are beginning to uncover clear evidence that elevated IOP produces progressive changes in the optic nerve head and retina. In the optic nerve head, these include axonal and non-axonal effects, the latter pointing to involvement of extracellular matrix and astrocyte responses. In the retina, retinal ganglion cells appear to undergo changes in neurotrophin response as well as morphologic changes prior to actual cell death. These, and other, as yet uncovered, abnormalities in the optic nerve head and retina may influence relative susceptibility to IOP and explain progressive optic nerve damage and visual field loss, in spite of apparent, clinically adequate IOP control. Ultimately, this knowledge may lead to the development of new treatments designed to preserve vision in these difficult patients.     

A new experimental model of glaucoma in rats through intracameral injections of hyaluronic acid

An experimental model of pressure-induced optic nerve damage would greatly facilitate the understanding of the cellular events leading to ganglion cell death, and how they are influenced by intraocular pressure and other risk factors associated to glaucoma. The aim of the present report was to study the effect of a long-term increase of intraocular pressure in rats induced by intracameral injections of hyaluronic acid with respect to electroretinographic activity and retinal and optic nerve histology. For this purpose, hyaluronic acid was injected weekly in the rat anterior chamber of one eye, whereas the contralateral eye was injected with saline solution. The results showed a significant decrease of oscillatory potentials and a- and b-wave amplitude of the scotopic electroretinogram after 3 or 6 weeks of hyaluronic acid administration, respectively. These parameters were further reduced after 10 weeks of treatment with hyaluronic acid. No significant changes in anterior chamber angle structures from hyaluronic acid- and vehicle-injected eyes were observed, whereas a significant loss of ganglion cell layer cells and of optic nerve axons were detected in animals that received hyaluronic acid for 10 weeks, as compared to eyes injected with saline solution. In summary, present results indicate that the chronic administration of hyaluronic acid induced a significant decrease in the electroretinographic activity and histological changes in the retina and optic nerve that seem consistent with some features of chronic open-angle glaucoma. Therefore, this could be an experimental model to study the cellular mechanisms by which elevated intraocular pressure damages the optic nerve and the retina.     

Neurotrophin roles in retinal ganglion cell survival: Lessons from rat glaucoma models

The neurotrophin (NT) hypothesis proposes that the obstruction of retrograde transport at the optic nerve head results in the deprivation of neurotrophic support to retinal ganglion cells (RGC) leading to apoptotic cell death in glaucoma. An important corollary to this concept is the implication that appropriate enhancement of neurotrophic support will prolong the survival of injured RGC indefinitely. This hypothesis is, perhaps, the most widely recognized theory to explain RGC loss resulting from exposure of the eye to elevated intraocular pressure (IOP). Recent studies of NT signaling using rat glaucoma models, have examined the endogenous responses of the retina to pressure exposure as well as studies designed to augment NT signaling in order to rescue RGC from apoptosis following pressure-induced injury. The examination of these studies in this review reveals a number of consistent observations and provides direction for further investigations of this hypothesis.     

The mechanism of optic nerve damage in experimental acute intraocular pressure elevation

We produced intraocular pressure (IOP) elevations in 32 primate eyes and studied retinal ganglion cell rapid axonal transport with autoradiography and electron microscopy. Animals breathing room air at sea level pressure were compared to animals breathing 100% oxygen at 3 atm pressure in a hyperbaric chamber. Despite major increases in arterial oxygen levels in the hyperbarically oxygenated animals, both groups had axonal transport blockade at the optic nerve head. Anoxia appears not to be the most important cause of acute axonal damage induced by elevated IOP. The pattern of axonal abnormality within individual fiber bundles at the optic nerve head provides support for mechanical compression as a more likely alternative cause for induced neural damage.     

Expression of myocilin/TIGR in normal and glaucomatous primate optic nerves

Myocilin/TIGR was the first molecule discovered to be linked with primary open angle glaucoma (POAG), a blinding disease characterized by progressive loss of retinal ganglion cells. Mutations in myocilin/TIGR have been associated with age of disease onset and severity. The function of myocilin/TIGR and its role in glaucoma is unknown. Myocilin/TIGR has been studied in the trabecular meshwork to determine a role in regulation of intraocular pressure. The site of damage to the axons of the retinal ganglion cells is the optic nerve head (ONH). The myocilin/TIGR expression was examined in fetal through adult human optic nerve as well as in POAG. Myocilin/TIGR was expressed in the myelinated optic nerve of children and normal adults but not in the fetal optic nerve before myelination. Also examined was the expression in monkeys with experimental glaucoma. The results demonstrate that optic nerve head astrocytes constitutively express myocilin/TIGR in vivo in primates. Nevertheless, myocilin/TIGR is apparently reduced in glaucomatous ONH. The colocalization of myocilin/TIGR to the myelin suggests a role of myocilin/TIGR in the myelinated optic nerve.     

Autophagy in axonal degeneration in glaucomatous optic neuropathy

The role of autophagy in retinal ganglion cell (RGC) death is still controversial. Several studies focused on RGC body death, although the axonal degeneration pathway in the optic nerve has not been well documented in spite of evidence that the mechanisms of degeneration of neuronal cell bodies and their axons differ. Axonal degeneration of RGCs is a hallmark of glaucoma, and a pattern of localized retinal nerve fiber layer defects in glaucoma patients indicates that axonal degeneration may precede RGC body death in this condition. As models of preceding axonal degeneration, both the tumor necrosis factor (TNF) injection model and hypertensive glaucoma model may be useful in understanding the mechanism of axonal degeneration of RGCs, and the concept of axonal protection can be an attractive approach to the prevention of neurodegenerative optic nerve disease. Since mitochondria play crucial roles in glaucomatous optic neuropathy and can themselves serve as a part of the autophagosome, it seems that mitochondrial function may alter autophagy machinery. Like other neurodegenerative diseases, optic nerve degeneration may exhibit autophagic flux impairment resulting from elevated intraocular pressure, TNF, traumatic injury, ischemia, oxidative stress, and aging. As a model of aging, we used senescence-accelerated mice to provide new insights. In this review, we attempt to describe the relationship between autophagy and recently reported noteworthy factors including Nmnat, ROCK, and SIRT1 in the degeneration of RGCs and their axons and propose possible mechanisms of axonal protection via modulation of autophagy machinery.     

Glaucoma as primary optic neuropathy

The appearance of the new concept of glaucomatous optic neuropathy directed to idea that high intraocular pressure and vascular insufficiency in the optic nerve head are only risk factors in the appearance of glaucoma. Glaucomatous optic neuropathy is the consequence of progressive loss of the retinal ganglion cells whose axons comprise the optic nerve. A great number of neurotrophic agents are under investigation with the view of preventing the dead of retinal ganglion cells. Induction of cell rescue mechanisms may be an alternate and efficient strategy for neuroprotection. The paper presents both actual data about the pathophysiological mechanisms of glaucoma (neurotrophin deprivation, excitotoxicity caused by glutamate, "ischemic cascade", apoptosis of retinal ganglion cells) and neuroprotective and neuroregenerative therapy and antiapoptotic agents.     

Neuronal death in glaucoma

Glaucoma is recognized to have its major detrimental effect upon the eye by killing retinal ganglion cells. The process of cell death appears to be initiated at the optic nerve head, though other sites of injury are possible but unsubstantiated. At present the injury at the nerve head seems related to the level of the eye pressure, but its detailed mechanism is as yet unexplained. There is a greater loss of ganglion cells from some areas of the eye, and this feature of glaucoma seems related to the regional structure of the supporting connective tissues of the optic nerve head. Larger retinal ganglion cells have been consistently shown to have somewhat greater susceptibility to injury in glaucoma, though all cells are injured, even early in the process. Ganglion cells die by apoptosis in human and experimental glaucoma, opening several potential areas for future therapies to protect them from dying. Neurotrophin deprivation is one possible cause of cell death and replacement therapy is a potential approach to treatment.     

Nitric Oxide: A Potential Mediator of Retinal Ganglion Cell Damage in Glaucoma

In the glaucomatous optic nerve head, excessive nitric oxide (NO) may be responsible, at least in part, for degeneration of axons of retinal ganglion cells. We have demonstrated the apparent up-regulation and induction of certain isoforms of nitric oxide synthase (NOS), the enzyme that synthesizes NO, in astrocytes in the prelaminar and lamina cribrosa regions of optic nerve heads of patients with glaucoma. Evidence of NO toxicity, histochemical staining for nitrotyrosine, is present in these damaged optic nerve heads. In rats with experimentally induced, moderately elevated intraocular pressure, the isoforms of NOS were also identified.     

Neuroprotektion bei Glaukom bleibt ein Konzept

According to estimates made by WHO, approximately 105 million people are affected worldwide by glaucoma. This can be defined as progressive optic neuropathy with structural damage of the optic nerve head and death of retinal ganglion cells. Although elevated IOP is considered responsible for glaucoma, lowering the pressure often does not result in improvement. For this reason, other etiological factors are presumed, which are presented in the following contribution. The role of neuroprotective agents in the treatment of glaucoma is discussed. The pattern of ganglion cell death specific to glaucoma seems to suggest that certain ganglion cells could be more sensitive than others. The theory of "cumulative damage" in this case includes the hypothesis that the delayed onset of many neurodegenerative diseases such as glaucoma, Alzheimer's disease, or Parkinson's disease can be attributed to the age-related accumulation of toxic substances in the ganglion cells. On the contrary, the theory of "singular damage" is based on the assumption that certain ganglion cells are in a state of reduced homeostasis caused by the expression of so-called mutant response genes. Therapeutic approaches worthy of consideration based on their side effect profile and efficacy in animal trials, are presented.     

Biomechanics of the sclera and effects on intraocular pressure

Accumulating evidence indicates that glaucoma is a multifactorial neurodegenerative disease characterized by the loss of retinal ganglion cells (RGC), resulting in gradual and progressive permanent loss of vision. Reducing intraocular pressure (IOP) remains the only proven method for preventing and delaying the progression of glaucomatous visual impairment. However, the specific role of IOP in optic nerve injury remains controversial, and little is known about the biomechanical mechanism by which elevated IOP leads to the loss of RGC. Published studies suggest that the biomechanical properties of the sclera and scleral lamina cribrosa determine the biomechanical changes of optic nerve head, and play an important role in the pathologic process of loss of RGC and optic nerve damage. This review focuses on the current understanding of biomechanics of sclera in glaucoma and provides an overview of the possible interactions between the sclera and IOP. Treatments and interventions aimed at the sclera are also discussed.     

The optic nerve head in normal-tension glaucoma

There is controversy over the definition, appearance, and characteristics of the optic nerve head in normal-tension glaucoma (NTG). Optic disk size is greater in eyes with NTG than in those with primary open-angle glaucoma. However, in an intraindividual bilateral comparison, the eye with the larger optic disk showed neither more marked nor less pronounced glaucomatous optic nerve damage. Optic disk hemorrhage and peripapillary atrophy have been reported to be more frequent in patients with NTG. Nonuse of calcium channel blockers, peripapillary atrophy, and disk hemorrhage were statistically significantly associated with visual field loss progression in NTG. However, there is a possibility that a high IOP may stop disk hemorrhage relatively early. Histopathologic optic nerve head changes correlated with the clinical appearance of the optic nerve head, which is comparable in NTG and primary open-angle glaucoma. However, as novel findings, serum antibodies to retinal proteins and retinal immunoglobulin deposition in the ganglion cells were observed, and the level of serum autoantibodies to optic nerve head glycosaminoglycans was higher in patients with NTG than in patients with primary open-angle glaucoma.     

Clinical value of scanning laser polarimetry in glaucoma diagnostics

The term glaucoma is used as a melting pot of many different diseases which have in common that the retinal ganglion cells and their axons are damaged. Untreated, apoptosis can be induced causing ganglion cell death which subsequently leads to typical glaucomatous damage at the optic nerve head, scotomas of the visual fields, and in the worst case scenario to blindness. It is well known that patients with glaucoma can suffer a 20 to 50?% loss of retinal ganglion cells before a defect becomes evident in standard white on white perimetry. To prevent glaucomatous damage, it is important to detect changes of the retinal ganglion cells and their nerve fibre layer as early as possible and to monitor their follow-up as closely as possible in order to find an adequate treatment of glaucoma, and to control its efficiency. In the past few years, scanning laser polarimetry by means of GDx technology (Carl Zeiss Meditec, Dublin, USA) could be established as a new method to measure the retinal nerve fibre layer not only qualitatively but even quantitatively. Presently, the GDx plays an important role in actual glaucoma diagnostics on account of its high resolution, the comfort for both patient and user, and its highly reproducible measurements. Especially in difficult evaluable optic nerve heads (e.?g., micro- and macrodiscs), tilted discs, and optic disc anomalies (e.?g., optic nerve drusen) modern nerve fibre diagnostics by means of GDx technology is a helpful enrichment in clinical routine.     

Glial cell and glaucomatous optic neuropathy

Glaucomatous optic neuropathy is a chronic disease accompanied by visual field loss, cupping of optic nerve head, and apoptosis of retinal ganglion cells (RGCs). The mechanism of glaucomatous optic neuropathy is unknown but glial cells play an important role in glaucomatous optic nerve damage and the repair process. We review the role of glial cells in the remodeling of optic nerve head, apoptosis of RGCs and immune reactions in glaucoma.     

Consequences of glaucoma on circadian and central visual systems

Glaucoma is a chronic optic neuropathy leading to a degeneration of retinal ganglion cells. There is accumulating evidence that glaucomatous damage extends from retinal ganglion cells to vision centers in the brain. Degenerative changes are observed in magnocellular, parvocellular, and koniocellular pathways in the lateral geniculate nucleus, and these changes are related to intraocular pressure and the severity of optic nerve damage. In addition, recent studies show that there are also changes in the visual cortex in relation to varying degrees of retinal ganglion cell loss. In a rat model of glaucoma, we have recently demonstrated a reduction of retinal projections of retinal ganglion cells, not only on the visual system but also on the suprachiasmatic nucleus. Human studies suggest that the ganglion cell degeneration caused by glaucoma could lead to a lesion of the retinohypothalamic tract, which permits the synchronization of circadian rhythms.     

Inherited glaucoma in DBA/2J mice: pertinent disease features for studying the neurodegeneration

The glaucomas are neurodegenerative diseases involving death of retinal ganglion cells and optic nerve head excavation. A major risk factor for this neurodegeneration is a harmfully elevated intraocular pressure (IOP). Human glaucomas are typically complex, progressive diseases that are prevalent in the elderly. Family history and genetic factors are clearly important in human glaucoma. Mouse studies have proven helpful for investigating the genetic and mechanistic basis of complex diseases. We previously reported inherited, age-related progressive glaucoma in DBA/2J mice. Here, we report our updated findings from studying the disease in a large number of DBA/2J mice. The period when mice have elevated IOP extends from 6 months to 16 months, with 8-9 months representing an important transition to high IOP for many mice. Optic nerve degeneration follows IOP elevation, with the majority of optic nerves being severely damaged by 12 months of age. This information should help with the design of experiments, and we present the data in a manner that will be useful for future studies of retinal ganglion cell degeneration and optic neuropathy.     

Should we treat the brain in glaucoma?

The loss of retinal ganglion cells in glaucoma may lead to blindness, and current therapies are directed at reducing pressure within the eye. Most of the retinal ganglion cell axon lies outside the eye, and evidence from experimental primate and human glaucoma suggests that axon injury extends from the optic nerve to visual pathways in the brain. Neurodegenerative changes in the central visual system may contribute to the pathology of glaucomatous progression. Thus, intraocular pressure-lowering strategies combined with neuroprotective therapies to protect visual neurons in the retina and brain may help to preserve vision in patients with glaucoma.     

Potential Role of Nitric Oxide and Endothelin in the Pathogenesis of Glaucoma

Glaucoma is an optic nerve head neuropathy in which retinal ganglion cells are lost. A clear association exists between glaucoma and different risk factors, such as high intraocular pressure (IOP) or blood-flow dysregulation. Nitric oxide (NO) and endothelin, two recently identified cellular mediators, appear to be involved in the regulation of IOP as well as in the modulation of ocular blood flow. To some extent, NO is also involved in apoptosis, a mechanism of cell death that can lead to retinal ganglion cell loss in glaucoma. This article provides a short and simplified overview of the biochemistry of NO and endothelin and highlights the potential role of these two mediators in certain important aspects related to the pathogenesis of glaucoma.