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Variable Partial Unilateral Ureteral Obstruction and Its Release in the Neonatal and Adult Mouse

Obstructive nephropathy is the most important cause of renal failure in children. Unilateral ureteral obstruction (UUO) in the neonatal mouse provides a useful model to investigate the response of the developing kidney to urine flow obstruction. Creation of reversible variable par ...

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Ultrasound Imaging of the Murine Kidney

Ultrasound (US) is the most common and least invasive modality for clinical imaging of the kidney. One important application of US in nephrology is the detection and monitoring of structural changes in the kidney. Recent advances in US technology have facilitated the application of similar ...

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Urinary Diversion via Cutaneous Vesicostomy in the Megabladder Mouse

Lower urinary tract obstruction in mice can lead to end-stage renal disease and death. We have developed a surgical technique to create a cutaneous vesicostomy in mice providing an external outlet for drainage of urine, thereby relieving the obstruction and slowing and/or preventing the de ...

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Assessment of Mitochondrial Damage in Retinal Cells and Tissues Using Quantitative Polymerase Chain Reaction for Mitochondrial DNA Damage and Extracel

Mitochondrial dysfunction and genomic instability are associated with a number of retinal pathologies including age-related macular degeneration, diabetic retinopathy, and glaucoma. Consequences of mitochondrial dysfunction within cells include elevation of the ra ...

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High-Throughput RNA In Situ Hybridization in Mouse Retina

The introduction of large-scale gene expression profiling studies has greatly increased the need to rapidly obtain high-quality cellular expression patterns of genes found to exhibit differential expression. The use of large-scale nonradioactive RNA in situ hybridization ma ...

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Detection of DNA Fragmentation in Retinal Apoptosis by TUNEL

Terminal dUTP nick end labeling (TUNEL) is an invaluable technique used in the study of late-stage apoptosis. The technique is based upon detection of fragmented DNA, a well-recognized characteristic of apoptosis, usually with fluorescent markers. Here, we describe the TUNEL technique ...

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A Method of Horizontally Sliced Preparation of the Retina

Various types of retinal neurons, including amacrine, ganglion, and horizontal cells, expand neurites (dendrites or axons) in horizontal direction and make synaptic or electrical contacts with other cells to integrate the visual information. Many types of ion-channels and recept ...

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Cell-Specific Markers for the Identification of Retinal Cells by Immunofluorescence Microscopy

Identification and visualization of specific cells and cellular structures in the retina are fundamental for understanding the visual process, retinal development, disease progression, and therapeutic intervention. The increased usage of transgenic and naturally occur ...

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Analysis of Photoreceptor Degeneration in the Zebrafish Danio rerio

Disturbances in the general mRNA metabolism have been recognized as a major defect in a growing number of hereditary human diseases. One prominent example of this disease group is Retinitis pigmentosa (RP), characterized by selective loss of photoreceptor cells. RP can be caused by dominant ...

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Generation of Transgenic X. laevis Models of Retinal Degeneration

Transgenic models are invaluable tools for researching retinal degenerative disease mechanisms. However, they are time-consuming and expensive to generate and maintain. We have developed an alternative to transgenic rodent models of retinal degeneration using transgenic X ...

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Analysis of the Drosophila Compound Eye with Light and Electron Microscopy

The Drosophila compound eye is a regular structure, in which about 750 units, called ommatidia, are arranged in a highly regular pattern. Eye development proceeds in a stereotypical fashion, where epithelial cells of the eye imaginal discs are specified, recruited, and differentiated in a s ...

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Analysis of Optokinetic Response in Zebrafish by Computer-Based Eye Tracking

Large-field movements in the visual surround trigger spontaneous, compensatory eye movements known as optokinetic response (OKR) in all vertebrates. In zebrafish (Danio rerio) the OKR is well developed at 5 days post fertilization and can be used in the laboratory for screening of visual p ...

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N -Methyl-d-Aspartate (NMDA)-Mediated Excitotoxic Damage: A Mouse Model of Acute Retinal Ganglion Cell Damage

The animal model of N-methyl-d-aspartate (NMDA)-induced excitotoxic damage of retinal ganglion cells (RGC) is widely used to study the molecular mechanisms of RGC apoptosis and/or its prevention by neuroprotective agents. This chapter provides protocols for applying NMDA-indu ...

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Light Damage as a Model of Retinal Degeneration

The induction of retinal degeneration by light exposure is widely used to study mechanisms of cell death. The advantage of such light-induced lesions over genetically determined degenerations is that light exposures can be manipulated according to the needs of the experimenter. Brig ...

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Phenotyping of Mouse Models with OCT

Optical coherence tomography (OCT) is an invaluable technique to perform noninvasive retinal imaging in small animal models such as mice. It provides virtual cross sections that correlate well with histomorphometric data with the advantage that multiple iterative measurements ...

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Functional Phenotyping of Mouse Models with ERG

In many situations it is important to be able to assess the degree of retinal function, e.g., for the characterization of mouse models with unknown retinal involvement, when studying degenerative processes, for the analysis of visual signal processing, and during the follow-up of therapeut ...

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Retinal Fundus Imaging in Mouse Models of Retinal Diseases

The development of in vivo retinal fundus imaging in mice has opened a new research horizon, not only in ophthalmic research. The ability to monitor the dynamics of vascular and cellular changes in pathological conditions, such as neovascularization or degeneration, longitudinally wi ...

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Mouse Models for Studies of Retinal Degeneration and Diseases

Mouse models, with their well-developed genetics and similarity to human physiology and anatomy, serve as powerful tools with which to investigate the etiology of human retinal degeneration. Mutant mice also provide reproducible, experimental systems for elucidating pathways ...

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Identification and Analysis of Inherited Retinal Disease Genes

Inherited retinal diseases display a very high degree of clinical and genetic heterogeneity, which poses challenges in identifying the underlying defects in known genes and in identifying novel retinal disease genes. Here, we outline the state-of-the-art techniques to find the causa ...

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Analysis of Photoreceptor Rod Outer Segment Phagocytosis by RPE Cells In Situ

Counting rhodopsin-positive phagosomes residing in the retinal pigment epithelium (RPE) in the eye at different times of day allows a quantitative assessment of engulfment and digestion phases of diurnal RPE phagocytosis, which efficiently clears shed photoreceptor outer seg ...

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