Annexin V binding assay as a tool to measure apoptosis in differentiated neuronal cells.
We describe a rapid and reliable method to quantitate the extent of apoptosis in neuronal cell cultures. Based on their annexin V-affinity, resulting from phosphatidylserine (PS) exposure at the outer leaflet of the plasma membrane, apoptotic cells can be distinguished from annexin V-negative living cells, by using microscopic and flow cytometric procedures. When combined with propidium iodide (PI) the double labeling procedure allows a further distinction of necrotic (annexin V+/PI+), apoptotic (annexin V+/PI-) cells. Furthermore, when the cells are incubated with annexin V prior to harvesting, the former cell populations can be separated from cells damaged during isolation (annexin V-/PI+). In the present paper, we show that the annexin V-binding assay is also applicable to differentiated neuronal cells with fragile neurite outgrowths.
Schutte
B
B
Department of Molecular Cell Biology and Genetics, University of Maastricht, The Netherlands. bert.schutte@molcelb.unimaas.nl
Nuydens
R
R
Geerts
H
H
Ramaekers
F
F
D017304:Annexin A5 / Q000378:metabolism*; D017209:Apoptosis / Q000235:genetics / Q000502:physiology; D004791:Enzyme Inhibitors / Q000378:metabolism; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D009474:Neurons / Q000502:physiology*; D011687:Purines / Q000378:metabolism; D000077546:Roscovitine; D014407:Tumor Cells, Cultured
D017304:Annexin A5; D004791:Enzyme Inhibitors; D011687:Purines; D000077546:Roscovitine