Visualization of cell death in vivo with the annexin A5 imaging protocol

Reutelingsperger, Chris; Dumont, Ewald A. W. J.; Thimister, P. W. L.; Genderen, Hugo van; Kenis, Heidi; Eijnde, Stefan van de; Heidendal, G. A. K.; Hofstra, Leo

doi:10.1016/s0022-1759(02)00075-3

Cited by 56 publications

(35 citation statements)

References 36 publications

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“…Furthermore, Cy3-annexin V revealed phosphatidylserine exposure on the external leaflet of the cytoplasmic membrane (10/10 cells; right inset in Fig. 3C), a typical early indicator of the activation of a death process (Reutelingsperger et al, 2002).…”

Section: E-atp Causes Death Of the Cholinergic Neurons In Normal Retimentioning

confidence: 98%

Neuronal death induced by endogenous extracellular ATP in retinal cholinergic neuron density control

et al. 2005

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The precise assembly of neuronal circuits requires that the correct number of pre- and postsynaptic neurons form synaptic connections. Neuronal cell number is thus tightly controlled by cell death during development. Investigating the regulation of cell number in the retina we found an ATP gated mechanism of neuronal death control. By degrading endogenous extracellular ATP or blocking the P2X7 ATP receptors we found that endogenous extracellular ATP triggers the death of retinal cholinergic neurons during normal development. ATP-induced death eliminates cholinergic cells too close to one another, thereby controlling the total number, the local density and the regular spacing of these neurons.

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Section: E-atp Causes Death Of the Cholinergic Neurons In Normal Retimentioning

confidence: 98%

Neuronal death induced by endogenous extracellular ATP in retinal cholinergic neuron density control

et al. 2005

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“…Lamin is a nuclear envelop protein and is cleaved by the interleukin-converting enzyme family during apoptosis (29). Annexins, structural proteins exhibiting Ca 2ϩ -dependent binding activity to phospholipids, are known biomarkers for apoptosis (30). Voltage-dependent anion channel is a major component of the permeability transition pore complex of the mitochondrial megachannel and is associated with members of the Bcl-2 family (31).…”

Section: Proteomic Analysis Of the Scnt Extraembryonic Tissue And Idementioning

confidence: 99%

Proteomic Analysis of the Extraembryonic Tissue from Cloned Porcine Embryos

Chae

Cho

Seo

et al. 2006

Molecular & Cellular Proteomics

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Cloned animals developed from somatic cell nuclear transfer (SCNT) embryos are useful resources for agricultural and medical applications. However, the birth rate in the cloned animals is very low, and the cloned animals that have survived show various developmental defects. In this report, we present the morphology and differentially regulated proteins in the extraembryonic tissue from SCNT embryos to understand the molecular nature of the tissue. We examined 26-day-old SCNT porcine embryos at which the sonogram can first detect pregnancy. The extraembryonic tissue from SCNT embryos was abnormally small compared with the control. In the proteomic analysis with the SCNT extraembryonic tissue, 39 proteins were identified as differentially regulated proteins. Among up-regulated proteins, Annexins and Hsp27 were found. They are closely related to the processes of apoptosis. Among down-regulated proteins, Peroxiredoxins and anaerobic glycolytic enzymes were identified. In the Western blot analysis, antioxidant enzymes and the antiapoptotic Bcl-2 protein were down-regulated, and caspases were up-regulated. In the terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling (TUNEL) assay with the placenta from SCNT embryos, apoptotic trophoblasts were observed. These results demonstrate that a major reason for the low birth rate of cloned animals is due to abnormal apoptosis in the extraembryonic tissue during early pregnancy. Molecular & Cellular Proteomics 5:1559 -1566, 2006.

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“…If we could directly visualize this process, it would facilitate a much more precise diagnosis and critically enable accurate tracking of the disease state and the action of therapy. However, until now, it has not been possible to detect nerve cell apoptosis in vivo (1,(7)(8)(9)(10).…”

mentioning

confidence: 99%

“…However, existing in vivo techniques using annexin 5 either have been unable to resolve the process to a single cellular level (7)(8)(9) or require an invasive method performed under terminal anesthesia (10). Imaging the eye, compared with the rest of the body, offers a unique opportunity because of the presence of clear optical media allowing direct visualization of labeled disease processes as they occur.…”

mentioning

confidence: 99%

Real-time imaging of single nerve cell apoptosis in retinal neurodegeneration

Cordeiro

Guo

Luong

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

244

308

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Apoptotic nerve cell death is implicated in the pathogenesis of several devastating neurodegenerative conditions, including glaucoma and Alzheimer's and Parkinson's diseases. We have devised a noninvasive real-time imaging technique using confocal laserscanning ophthalmoscopy to visualize single nerve cell apoptosis in vivo, which allows longitudinal study of disease processes that has not previously been possible. Our method utilizes the unique optical properties of the eye, which allow direct microscopic observation of nerve cells in the retina. We have been able to image changes occurring in nerve cell apoptosis over hours, days, and months and show that effects depend on the magnitude of the initial apoptotic inducer in several models of neurodegenerative disease in rat and primate. This technology enables the direct observation of single nerve cell apoptosis in experimental neurodegeneration, providing the opportunity for detailed investigation of fundamental disease mechanisms and the evaluation of interventions with potential clinical applications, together with the possibility of taking this method through to patients.A poptosis is an orchestrated form of cell ''death by suicide.'' It is essential in both the development and normal maintenance of tissue function. It is also implicated in the pathology of a number of severe neurodegenerative disorders such as glaucoma, motor neuron, and Alzheimer's, Parkinson's, and Huntington's diseases (1). There is evidence that a similar pathogenesis may contribute to neurodegenerative processes in these conditions (2), and that the extent of nerve cell loss is correlated with functional deficit (3-6). If we could directly visualize this process, it would facilitate a much more precise diagnosis and critically enable accurate tracking of the disease state and the action of therapy. However, until now, it has not been possible to detect nerve cell apoptosis in vivo (1, 7-10).Annexin 5 is a protein that, in the presence of Ca 2ϩ , has a high affinity for phosphatidylserine (PS), an anionic phospholipid that is enriched in the inner leaflet of plasma membranes. The externalization of PS from the inner leaflet to the outer layer of the cell membrane is an invariant early feature in the apoptotic process that occurs before DNA fragmentation and nuclear condensation. Because of its properties, FITC-annexin 5 has become widely used in the cytological detection of cells undergoing apoptosis (11). More recently, annexin 5 has been shown to be effective in the identification of apoptosis in vivo by using radiological and macroscopic fluorescent techniques (7,8,10,12,13).However, existing in vivo techniques using annexin 5 either have been unable to resolve the process to a single cellular level (7-9) or require an invasive method performed under terminal anesthesia (10). Imaging the eye, compared with the rest of the body, offers a unique opportunity because of the presence of clear optical media allowing direct visualization of labeled disease processes as they occur. This mea...

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Visualization of cell death in vivo with the annexin A5 imaging protocol

Cited by 56 publications

References 36 publications

Neuronal death induced by endogenous extracellular ATP in retinal cholinergic neuron density control

Neuronal death induced by endogenous extracellular ATP in retinal cholinergic neuron density control

Proteomic Analysis of the Extraembryonic Tissue from Cloned Porcine Embryos

Real-time imaging of single nerve cell apoptosis in retinal neurodegeneration

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