Ethanol impairs microtubule formation via interactions at a microtubule associated protein-sensitive site

Katherine J. Smith, Tracy R. Butler, Mark A. Prendergast

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

Abstract

Prolonged ethanol abuse has been associated with brain injury caused by impaired synaptogenesis, cellular migration, neurogenesis, and cell signaling, all of which require proper microtubule functioning. However, the means by which ethanol may impair microtubule formation or function and the role that microtubule-associated proteins (MAPs) have in mediating such effects are not clear. In the present studies, purified MAP-deficient (2mg/mL) and MAP-rich (pre-conjugated; 1mg/mL) bovine α/β tubulin dimer was allowed to polymerize at 37°C, forming microtubules in the presence or absence of ethanol (25-500m. m). Microtubule formation was assessed in a 96-well format using a turbidity assay, with absorption measured at 340nm for 45min. Additional studies co-exposed α/β tubulin dimers to 50m. m ethanol and purified MAPs (0.1mg/mL) for 45min. Polymerization of MAP-deficient tubulin was significantly decreased (at 15-45min of polymerization) during exposure to ethanol (>25m. m). In contrast, ethanol exposure did not alter polymerization of α/β tubulin dimers pre-conjugated to MAPs, at any concentration. Concurrent exposure of MAP-deficient tubulin with purified MAPs and ethanol resulted in significant and time-dependent decreases in tubulin polymerization, with recovery from inhibition at later time points. The present results suggest that ethanol disrupts MAP-independent microtubule formation and MAP-dependent microtubule formation via direct actions at an MAP-sensitive microtubule residue, indicating that disruption of neuronal microtubule formation and function may contribute to the neurodegenerative effects of binge-like ethanol intake.

Original languageEnglish
Pages (from-to)539-543
Number of pages5
JournalAlcohol
Volume47
Issue number7
DOIs
StatePublished - Nov 2013

Bibliographical note

Funding Information:
The authors would like to acknowledge the support of AA013566, AA013388, and DA016176, from the National Institute on Alcohol Abuse and Alcoholism and the National Institute on Drug Abuse.

Keywords

  • Alcoholism
  • Neuronal injury
  • Polymerization
  • Tubulin

ASJC Scopus subject areas

  • Health(social science)
  • Biochemistry
  • Toxicology
  • Neurology
  • Behavioral Neuroscience

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