Colchicine-induced cytoskeletal collapse and apoptosis in N-18 neuroblastoma cultures is rapidly reversed by applied S-100beta

L. S. Brewton, L. Haddad, E. C. Azmitia

Research output: Contribution to journalArticle

Abstract

Brain connections depend on a stable association between dendrites and axons whose cytoskeleton is stabilized by the proteins MAP-2 and tau, respectively. The glial protein S-100beta inhibits the phosphorylation by PKC of these two microtubule-associated proteins. In order to determine if exogenous S-100beta can directly influence the cytoskeleton of living cells, cultures of N-18 cells (neuroblastoma clonal cell line) are treated for 30 min in serum-free medium with 10(-6) M colchicine. In normal media, colchicine induces a rapid retraction of processes, membrane blebbing, nuclear collapse, and cell death. The observed cellular changes, due to cytoskeletal collapse after exposure to colchicine, are similar and consistent with the loss of processes and cytoplasmic blebbing seen in cells undergoing apoptosis. The addition of 20 ng/ml of S-100beta after the initial 30-min exposure to colchicine prevents apoptosis, nuclear collapse and induces the regrowth of retracted processes. Cells were treated with the Hoechst Stain, a fluorescent marker that binds to nuclear material, to determine the occurrence of apoptosis in our cultures. In our control cultures, receiving no drugs, we found that 15.1% of the cells were apoptotic. When colchicine was added to the culture medium we found that 31.6% of the cells became apoptotic. However, when colchicine was followed by exposure to S-100beta we found that only 5.4% of the cells were apoptotic. Our results suggest that extracellular application of the glial protein S-100beta is sufficient to reverse colchicine-induced cytoskeletal collapse and prevent the resultant apoptosis of the cells. The increased levels of S-100beta seen after brain injury and in certain neurological and psychiatric disorders may be considered as beneficial for brain recovery.
Original languageUndefined
Pages (from-to)9-16
Number of pages8
JournalBrain Research
Volume912
Issue number1
StatePublished - 2001

Keywords

  • Apoptosis/*drug effects/physiology Benzimidazoles/pharmacokinetics Cell Division/physiology Cell Survival/drug effects/physiology Colchicine/*antagonists & inhibitors Culture Media, Serum-Free/pharmacology Cytoskeleton/*drug effects/metabolism/pathology Drug Interactions/physiology Fluorescent Dyes/pharmacokinetics Humans Microscopy, Video Nerve Degeneration/drug therapy/physiopathology/*prevention & control Nerve Growth Factors Nerve Regeneration/*drug effects/physiology Neuroblastoma Neurons/*drug effects/metabolism/pathology S100 Calcium Binding Protein beta Subunit S100 Proteins/metabolism/*pharmacology Tumor Cells, Cultured/*drug effects/metabolism/pathology

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