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

Luke S. Brewton, Lisa Haddad, Efrain 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-100β inhibits the phosphorylation by PKC of these two microtubule-associated proteins. In order to determine if exogenous S-100β 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-100β 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-100β we found that only 5.4% of the cells were apoptotic. Our results suggest that extracellular application of the glial protein S-100β is sufficient to reverse colchicine-induced cytoskeletal collapse and prevent the resultant apoptosis of the cells. The increased levels of S-100β seen after brain injury and in certain neurological and psychiatric disorders may be considered as beneficial for brain recovery.

Original languageEnglish (US)
Pages (from-to)9-16
Number of pages8
JournalBrain Research
Volume912
Issue number1
DOIs
StatePublished - Aug 31 2001

Fingerprint

Colchicine
Neuroblastoma
Apoptosis
S100 Proteins
Blister
Cytoskeleton
Neuroglia
Microtubule-Associated Proteins
Serum-Free Culture Media
Nuclear Envelope
Brain
Dendrites
Nervous System Diseases
Brain Injuries
Psychiatry
Axons
Culture Media
Cell Death
Coloring Agents
Cell Culture Techniques

Keywords

  • Cytoskeleton
  • Hoechst stain
  • Microtubules
  • Morphometrics
  • Time-lapse
  • Tubulin

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Colchicine-induced cytoskeletal collapse and apoptosis in N-18 neuroblastoma cultures is rapidly reversed by applied S-100β. / Brewton, Luke S.; Haddad, Lisa; Azmitia, Efrain C.

In: Brain Research, Vol. 912, No. 1, 31.08.2001, p. 9-16.

Research output: Contribution to journalArticle

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