Ultrastructural consequences of radical damage before and after differentiation of neuroblastoma B-104 cells.
There is abundant evidence that the pathophysiology leading to neuronal death during post-ischemic brain reperfusion involves radical-mediated damage. Although the ultrastructural alterations accompanying brain ischemia and reperfusion are well characterized, little is known about the ultrastructural alterations that are specific to radical damage. This study examines in differentiated and undifferentiated neuroblastoma B-104 cells the viability (by dye exclusion) and ultrastructural consequences of radical damage initiated by 50 microM cumene hydroperoxide (CumOOH). Differentiation was most notably associated with formation of neurites and an extensive cytoskeletal feltwork. CumOOH-induced cell death was increased after differentiation and was blocked by the iron chelator DETAPAC. The ultrastructural characteristics of radical damage here included: (1) plasmalemmal holes that appear to undergo "patching" by well-organized membrane whorls, (2) accumulation of numerous free ribosomes, (3) markedly increased vesicular trafficking about the Golgi accompanied by Golgi transformation from cisternal organization to clusters of vacuoles with numerous fusing vesicles, (4) development of large multi-layered vacuoles that include damage membranes and organelles and appear to undergo extrusion from the cell, and (5) a general loss of cytoplasmic volume. These ultrastructural alterations developed more rapidly and were consistently more advanced in differentiated cells throughout the 6-h time course. In differentiated cells radical damage also induced the disorganization and subsequent loss of the extensive feltwork of cytoskeletal elements. There was little damage to the membranes of the nuclear envelope and mitochondria. Our observations in this system are strikingly similar to ultrastructural alterations in Golgi and ribosomal organization seen in vulnerable neurons during post-ischemic brain reperfusion and suggest that these alterations during reperfusion reflect the consequence of radical-mediated damage.
O'Neil
B J
BJ
Emergency Department, Detroit Receiving Hospital, MI 48201, USA.
Alousi
S S
SS
White
B C
BC
Rafols
J A
JA
D000818:Animals; D001555:Benzene Derivatives / Q000633:toxicity*; D001932:Brain Neoplasms / Q000473:pathology / Q000648:ultrastructure; D002454:Cell Differentiation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D005609:Free Radicals / Q000633:toxicity*; D009447:Neuroblastoma / Q000473:pathology* / Q000648:ultrastructure*; D016877:Oxidants / Q000633:toxicity*; D051381:Rats; D014407:Tumor Cells, Cultured
D001555:Benzene Derivatives; D005609:Free Radicals; D016877:Oxidants; C007164:cumene hydroperoxide