Name: a mouse model of retinal ischemia-reperfusion injury through elevation of intraocular pressure
Uploaded: 2016-07-14T14:00:00
Description: Watch this Scientific Journal Video about A Mouse Model of Retinal Ischemia-Reperfusion Injury Through Elevation of Intraocular Pressure at JoVE.com

This work was supported by research grants from the National Institutes of Health (EY022383 and EY022683; EJD) and Core grant (P30EY001765), Imaging and Microscopy Core Module.

Ethics Statement: All procedures were performed in accordance with the guidelines set forth by the Johns Hopkins University Institutional Animal Care and Use Committee.
Note: Mice used during filming are C57BL/6 mice from Jackson, although other rodent strains or species may also be used. When using other strains or species, be aware that anesthesia dosages and injury timeline may vary. It is important to adapt I/R conditions to accommodate strain, species, and experimental variations.
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<ol>
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Retinal I/R&#160;injury by elevated IOP has proven its utility in modeling cellular damage and dysfunction, particularly neurodegeneration, in the rodent retinal neurovascular unit. This procedure provides a robust control tissue and is easily accessible in terms of technical sophistication. It has been noted in this and other I/R injury models that increasing the pressure and duration of ischemia may increase injury severity24. For this reason, some practitioners have elected to use ischemic pressures and dur...

Retinal ischemia-reperfusion (I/R) characterizes many human retinal pathologies, including glaucoma, diabetic retinopathy, and retinal vascular occlusions1. In retinal I/R, reduced blood flow (ischemia) in the retinal vasculature creates a state of retinal hypersensitivity to oxygen and other nutrients, precipitating severe oxidative and inflammatory damage when circulation is subsequently reinstated (reperfusion)2. The neural retina appears particularly vulnerable to these changes, with retinal neurodegeneration being perhaps the most salient feature of I/R-induced damage. Presented here is a protocol for modeling retinal I/R injury in the mouse. This technique enables researchers to examine potential mechanisms and treatment strategies for human diseases of the retinal neurovascular unit.
Pioneered in 1952 by surgeons seeking to understand the neurodegenerative consequences of surgical anemia3, rodent retinal I/R by elevated intraocular pressure (IOP) was reestablished in 1991 for the purpose of standardizing neurodegenerative endpoints after ischemic insult4. Using the drip of a saline reservoir to raise IOP above systolic blood pressure, these studies demonstrated that pressurized ocular cannulation was sufficient to suspend the retinal circulation and thereby initiate neuronal degeneration. More recent efforts using retinal I/R by elevated IOP have begun to elaborate the mechanisms underlying I/R-induced retinal neurodegeneration5-12. Multiple groups have reported additional pathologic changes including inflammation13,14, vascular permeability15,16, and capillary degeneration14,17. Taken together, these studies have established retinal I/R injury by elevated IOP as a model of retinal neurovascular disease more generally.
Characterizing the mechanisms of I/R injury is essential for the study of vascular disease. Retinal I/R injury by elevated IOP is one of many hypoxia-induced injury models, including I/R injuries in lung18, heart19, brain20, liver21, kidney22, and intestine23. These models have been paramount in advancing our understanding of vascular illness and its clinical remedies. By extending the investigation of I/R processes to ocular tissues, retinal I/R injury by elevated IOP helps to paint a more comprehensive picture of these related conditions.
Corresponding closely with clinical neurodegenerative conditions in retina, retinal I/R injury by elevated IOP presents a valuable tool for researchers interested in exploring ischemic pathogenesis. The protocol described herein is targeted, tractable, and accessible. It is complemented well by endpoints in neuronal degeneration, such as quantification of retinal neurons, measurement of retinal thickness, and electrophysiological recording of retinal neuron function. This model has proven its utility in advancing neurovascular inquiry, and it shows promise in earning status as a foundational protocol in visual medicine research.

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a mouse model of retinal ischemia-reperfusion injury through elevation of intraocular pressure

Retinal ischemia-reperfusion (I/R) is a pathophysiological process contributing to cellular damage in multiple ocular conditions, including glaucoma, diabetic retinopathy, and retinal vascular occlusions. Rodent models of I/R injury are providing significant insights into mechanisms and treatment strategies for human I/R injury, especially with regard to neurodegenerative damage in the retinal neurovascular unit. Presented here is a protocol for inducing retinal I/R injury in mice through elevation of intraocular pressure (IOP). In this protocol, the ocular anterior chamber is cannulated with a needle, through which flows the drip of an elevated saline reservoir. Using this drip to raise IOP above systolic arterial blood pressure, a practitioner temporarily halts inner retinal blood flow (ischemia). When circulation is reinstated (reperfusion) by removal of the cannula, severe cellular damage ensues, resulting ultimately in retinal neurodegeneration. Recent studies demonstrate inflammation, vascular permeability, and capillary degeneration as additional elements of this model. Compared to alternative retinal I/R methodologies, such as retinal arterial ligation, retinal I/R injury by elevated IOP offers advantages in its&#160;anatomical specificity, experimental tractability, and technical accessibility, presenting itself as a valuable tool for examining neuronal pathogenesis and therapy in the retinal neurovascular unit.

The neurodegenerative effects of retinal I/R by elevated IOP are commonly evaluated using two standard approaches. NeuN immunolabeling of neuronal nuclei has revealed significant neuronal cell loss following I/R insult (Figure 1). Briefly, eyes enucleated 7 days after I/R were fixed in paraformaldehyde, labeled with the neuronal cell marker NeuN, and whole-mounted. Images were captured using confocal microscopy, and cells labeled with NeuN were quantified by counting...

Watch this Scientific Journal Video about A Mouse Model of Retinal Ischemia-Reperfusion Injury Through Elevation of Intraocular Pressure at JoVE.com

A Mouse Model of Retinal Ischemia-Reperfusion Injury Through Elevation of Intraocular Pressure

This article describes a procedure for inducing retinal ischemia-reperfusion injury by elevated intraocular pressure in mice. Retinal ischemia-reperfusion injury by elevated intraocular pressure serves to model human pathologies characterized by compromised oxygen and nutrient delivery in the retina, enabling researchers to examine potential cellular mechanisms and treatments for human diseases of the retinal neurovascular unit.

Retinal ischemia-reperfusion (I/R) is a pathophysiological process contributing to cellular damage in multiple ocular conditions, including glaucoma, ...

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