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丁香实验推荐阅读
Sensing BMP Pathway Activity by Immune Detection of Phosphorylated R-Smad Proteins in Mouse Embryonic Kidney

At the onset of mammalian kidney development, the ureteric bud invades the surrounding metanephric mesenchyme, and genetic studies in the mouse have shown that BMP pathway activity has to be antagonized in the vicinity of the epithelium, a task performed by the secreted BMP antagonist Grem1. H ...

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Immunohistochemical Staining of dpERK Staining During Early Kidney Development

Signaling through the ERK/MAPK pathway within the Wolffian duct and ureteric bud epithelium is critical for kidney induction and branching morphogenesis. ERK signaling is activated by receptor tyrosine kinase such as RET and FGFR2. This protocol describes a method to detect the diphos ...

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Analysis of In Vivo Transcription Factor Recruitment by Chromatin Immunoprecipitation of Mouse Embryonic Kidney

Chromatin immunoprecipitation (ChIP) is a powerful technique for examining transcription factor recruitment to chromatin, or histone modifications, at the level of specific genomic sequences. As such, it provides an invaluable tool for elucidating gene regulation at the molec ...

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IschemiaReperfusion Injury of the Mouse Kidney

Studies of the complex responses of the kidney to acute injury have yielded important insights into mechanisms of tissue injury and repair. A variety of injury models have contributed to this impressive body of knowledge, but the ischemia–reperfusion (IR) model has perhaps been the most wide ...

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Assessing Urinary Tract Defects in Mice: Methods to Detect the Presence of Vesicoureteric Reflux and Urinary Tract Obstruction

Congenital Anomalies of the Kidney and Urinary Tract (CAKUT) encompass a spectrum of kidney and urinary tract disorders. Here, we describe two assays that can be used to determine if a mouse has vesicoureteric reflux (VUR) or urinary tract obstruction, two urinary tract defects observed in CAK ...

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Estimating Total Nephron Number in the Adult Kidney Using the Physical Disector/Fractionator Combination

Nephron number has emerged as a useful parameter for assessing the roles of specific genes and feto-maternal environmental factors in kidney development. Nephron number is also of clinical interest due to increasing evidence suggesting that low nephron number is associated with incr ...

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Renal Delivery of Adenovirus and Antisense Oligonucleotides in Rats by Retrograde Renal Vein Injection

Renal gene therapy may offer new strategies to treat diseases of native and transplanted kidneys. Several experimental techniques have been developed using viral, nonviral, and cellular vectors, although the effectiveness of such techniques varies widely depending upon the vect ...

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Microinjection into the Lumen of the Ureteric Tree

During embryonic kidney development, the ureteric bud (UB) undergoes repetitive branching to generate the entire renal collecting system. Defects in UB branching result in renal malformations, from hypoplastic kidneys to renal agenesis. Mouse genetics has become an invaluable t ...

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Morpholino-Mediated Gene Knockdown in Mammalian Organ Culture

We examined the role of semaphorin3a in ureteric bud (UB) branching morphogenesis using mouse metanephric organ culture . In vitro UB injection of Sema3a antisense morpholino resulted in increased branching morphogenesis. Cellular and tissue uptake of oligonucleotides was faci ...

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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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