Annexin V binding

One of the early events of the apoptotic process involves the translocation of phosphatidylserine on the surface of cell membranes annexin V binding and propidium iodide uptake reveals various cellular states. After treatment with organotin(IV) compounds the cells could be categorized into populations vital cells (annexin V /P ), early apoptotic cells (annexin V /P ), late apoptotic cells (annexin V /P ), and necrotic cells (annexin V /P" ). Cells are observed with a fluorescence microscope and it is possible to observe translocation of phosphatidylserine (PS) from the inner side of the plasma membrane to the outer one and to see a green stain for annexin V FLUOS bound to PS, and a red stain for propidium iodide. 

CLARKE R G, LUND E K, JOHNSON I T aud PINDER A c (2000) Apoptosis cau he detected in attached colonic adenocarcinoma HT29 cells using annexin V binding, hut uot hy TUNEL assay or suh-GO DNA content . Cytometry, 39 141-50. 

Hwang and Bowen (2005b) Tomato paste hexane extract Apoptosis by Annexin V binding... 

Necrosis by LDH Mitochondrial transmembrane potential by DiOC6 and JC-1 fluorescence Apoptosis by cytochrome c release Annexin V binding DNA fragmentation by agarose gel electrophoresis... 

Annexin V is a human placental anticoagulant protein of molecular weight 35kDa that binds to membranes and lipid bilayers containing phosphatidylserine in the presence of free calcium. Annexin V binding to cell surfaces said to result from transmembrane movement of... 

Annexin V binding has been used to investigate the induction of apoptosis by a variety of factors. Induction of apoptosis in the human leukaemia cell hne, HL60, by photosensitizer dyes (Purpurin-18) administered in low doses after exposure to visible light, for example, has been monitored by annexin V binding to the cell surface (Di Stefano et al., 2001). It was found that when higher doses of the drug was employed necrosis of the cells resulted. 

Dillon, S.R., Mancini, M., Rosen, A., and Schlissel, M.S., 2000, Annexin V binds to viable B cells and colocahzed with a marker of lipid rafts upon C cell receptor activation, J. Immunol. 164 1322-1332. 

The combination of annexin V binding assay and the TUNEL method can reveal the presence of three subpopulations of apoptotic cells in tissues (1) annexin V-positive/TUNEL-negative cells which are in the early phase of apoptosis,... 

Fig. 11. Flow cytometric detection of apoptotic cell deadi using the annexin V method after treatment with IgM anti-Fas mAb. The vacant retrovirus vector (LXSN)-transfected Jurkat cell clone of LX-2 clearly exhibits annexin V binding at 3 h after treatment (56.8%), and the binding cell population is further increased at 5 h (77.2%), whereas only faint annexin V binding is observed in LdelSN-transfected Jurkat cell clone RJ-14. This means diat endogenous wild-type mFas in the LX-2 cells is functional in Fas-mediated apoptosis, but transfected aberrant mFas in die RJ-14 cells interferes with die signaUng in a dominant negative manner.

Simak J, Holada K, Vostal JG. Release of annexin V-binding membrane microparticles from cultured human umbilical vein endothelial cells after treatment with camptothecin. BM Cell Biol 2002 3 11. 

Treatment of the various tumor cell lines with cytotoxic anti-DNA autoantibodies induced internucleosomal DNA fragmentation and annexin V binding to the cell surface characteristic of the apoptotic pathway of cell death (K10). 

Homburg CH, de Haas M, von dem Borne AE, Verhoeven AJ, Reutelingsperger CP, Roos D. Human neutrophils lose their surface Fc gamma RIII and acquire Annexin V binding sites during apoptosis in vitro. Blood 1995 85 532-540. 

Merocyanine 540 is a red-emitting fluorescent dye that appears to be a sensitive measure of apoptosis-induced alterations in cytoplasmic membrane lipid structure (68). In apoptotic cells, the fluorescence increases in some models increased fluorescence correlates well with increased annexin V binding (68,71). Merocyanine 540 staining has been successfully used together with Hoechst 33342 to identify cell-cycle specific apoptotic events (72). 

Phosphatidylserine, which normally is on the inner leaflet of the plasma membrane, early during apoptosis becomes exposed on the outside cell surface (13). Because the anticoagulant protein annexin V binds with high affinity to phosphatidylserine, fluorochrome-conjugated annexin V can serve as a marker of apoptotic cells (21). During progression of apoptosis, the ability to bind annexin V precedes the loss of the plasma membrane s ability to exclude cationic dyes such as PI. 

Meers, R and Mealy, T, Phospholipid determinants for annexin V binding sites and the role of tryptophan 187, Biochemistry, 33[19], 5829, 1994. 

Caspase 3 upregulation, cyclin A levels, P53 level. Fas ligand expression, cell size and granularity (flow cytometric analysis), annexin V binding, DNA fragmentation, nuclei condensation... 

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