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细胞功能测定

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Detection and Measurement of Necrosis in Plants

Necrosis plays a fundamental role in plant physiology and pathology. When plants or plant cell cultures are subjected to abiotic stress they initiate rapid cell death with necrotic morphology. Likewise, when plants are attacked by pathogens, they develop necrotic lesions, the reaction ...

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Use of Necrotic Markers in the Drosophila Ovary

Necrosis is a form of cell death characterized by cytoplasmic and organelle swelling, compromised �membrane integrity, intracellular acidification, and increased levels of reactive oxygen species (ROS) and cytosolic Ca2+. In the Drosophila ovary, two distinct forms of cell death o ...

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Monitoring the Clearance of Apoptotic and Necrotic Cells in the Nematode Caenorhabditis elegans

The nematode Caenorhabditis elegans is an excellent model organism for studying the mechanisms �controlling cell death, including apoptosis, a cell suicide event, and necrosis, pathological cell deaths caused by environmental insults or genetic alterations. C. elegans has also b ...

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Necrosis in C. elegans

To use Caenorhabditis elegans to study the mechanisms for initiation and execution of necrosis, the experimentalist should be familiar with the established models of necrosis in C. elegans and the genetic and molecular tools available. We present a summary of two contrasting models for st ...

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Quantification of Genetically Controlled Cell Death in Budding Yeast

Yeast are the foremost genetic model system. With relative ease, entire chemical libraries can be screened for effects on essentially every gene in the yeast genome. Until recently, researchers focused only on whether yeast were killed by the conditions applied, irrespective of the mecha ...

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Identification and Characterization of Programmed Cell Death Markers in Bacterial Models

In eukaryotic organisms facing terminal stress, activation of genetically encoded cell death pathways underlies fundamental changes in core cellular processes and functional modification of critical biomolecules. These physiological alterations manifest themsel ...

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Quantitation of Acute Necrosis After Experimental Myocardial Infarction

Myocardial infarction (MI) is death and necrosis of myocardial tissue secondary to ischemia. MI is associated with adverse cardiac remodeling, progressive heart chamber dilation, ventricular wall thinning, and loss of cardiac function. Myocardial necrosis can be experimental ...

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Histological Analysis of Neurodegeneration in the Mouse Brain

Neurodegenerative diseases such as Alzheimer’s disease (AD) and Parkinson’s disease (PD) are characterized by chronic and progressive neuronal loss. Being able to detect and quantify neurodegeneration is the first step to identify mechanisms underlying neuronal cell death and to ...

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Analysis of Cell Death by Electron Microscopy

Cell death is said to occur mostly by two alternative, opposite modes: apoptosis, which involves a highly genetically regulated and elaborate network of biochemical events and cascades, and necrosis, considered a passive cell death without underlying regulatory mechanisms. Here, we ...

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Immunohistological Tools to Discriminate Apoptotic and Necrotic Cell Death in the Skin

Perturbances in skin homeostasis are responsible for the development of skin inflammatory diseases such as psoriasis or atopic dermatitis. While the role of apoptosis has been extensively studied in the skin, the role of the newly described programmed necrosis also termed necroptosis ...

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Analysis of Pyroptosis in Bacterial Infection

Eukaryotic cells undergo death by several different mechanisms: apoptosis, a cell death that prevents inflammatory response; necrosis, when the cell membrane lyses and all the intracellular content is spilled outside; and pyroptosis, a cell death that is accompanied by the release of in ...

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Methods for the Study of Entosis

Entosis is a recently described nonapoptotic cell death mechanism that is initiated by the engulfment of live epithelial cells, leading to the formation of “cell-in-cell” structures. Entotic cell engulfment is induced by matrix detachment, and is driven by imbalances in actomyosin con ...

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Fluorescent Biosensors for the Detection of HMGB1 Release

During necrosis and following some instances of apoptosis (in particular in the absence of a proficient phagocytic system), the nonhistone chromatin component high-mobility group box 1 (HMGB1) is released in the extracellular space. In vivo, extracellular HMGB1 can bind Toll-like re ...

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Activity Assays for Receptor-Interacting Protein Kinase 1:A Key Regulator of Necroptosis

Necroptosis is a novel form of regulated non-apoptotic cell death, which displays morphological features of necrosis. The kinase activity of receptor-interacting protein kinase 1 (RIP1) is a critical component in signaling for necroptosis. The development of assays to evaluate RIP1 ...

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Time-Lapse Imaging of Necrosis

The processes of dying are as tightly regulated as those of growth and proliferation. Recent work into the molecular pathways that regulate and execute cell death have uncovered a plethora of signalling cascades that lead to distinct modes of cell death, including “apoptosis,” “necrosis,” “ ...

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Navigation to the Graveyard-Induction of Various Pathways of Necrosis and Their Classification by Flow Cytometry

Apoptosis and necrosis reflect the program of cell death employed by a dying cell and the final stage of death, respectively. Whereas apoptosis is defined as a physiological, highly organized cell death process, necrosis is commonly considered to be accidental and uncontrolled. Physiol ...

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In Situ Hybridization of Radioactive Riboprobes to RNA in Tissue Sections

The development of such techniques as transgenesis, saturation mutagenesis, and the polymerase chain reaction (PCR), all of which are described in detail elsewhere in this volume, has revolutionized experimental embryology. However, no single procedure has been more broadly appl ...

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Gene Interference Using Antisense Oligodeoxynucleotides on Whole Chick Embryos: Optimal Ring and Roller-Bottle Culture Technique

Antisense oligodeoxynucleotides are being widely used to interfere with specific gene activities in cell-culture systems (1,2), and there are possible analytical advantages to partial and timed interference in the whole embryo, for certain genes, as compared with targeted null mut ...

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Protein Techniques: Immunoprecipitation, In Vitro Kinase Assays, and Western Blotting

The regulation of complex developmental pathways involves the spatial and temporal interaction of specific gene products. Several powerful genetic approaches, including the tagging of gene products or cells, have allowed their distribution in space and time to be determined. The ad ...

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Grafting Hensens Node

Soon after Spemann and Mangold’s (1) famous demonstration in 1924 that the dorsal lip of the blastopore of the gastrulating amphibian embryo has the unique ability to induce a second axis when grafted into an ectopic site in a host embryo, Waddington (2,3) showed that Hensen’s node is its equivalent in a ...

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