Cold exposure reveals two populations of microtubules in pulmonary endothelia

Cristhiaan D. Ochoa, Troy Stevens, Ron Balczon

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Microtubules are composed of α-tubulin and β-tubulin dimers. Microtubules yield tubulin dimers when exposed to cold, which reassemble spontaneously to form microtubule fibers at 37°C. However, mammalian neurons, glial cells, and fibroblasts have cold-stable microtubules. While studying the microtubule toxicity mechanisms of the exotoxin Y from Pseudomonas aeruginosa in pulmonary microvascular endothelial cells, we observed that some endothelial microtubules were very difficult to disassemble in the cold. As a consequence, we designed studies to test the hypothesis that microvascular endothelium has a population of cold-stable microtubules. Pulmonary microvascular endothelial cells and HeLa cells (control) were grown under regular cell culture conditions, followed by exposure to an ice-cold water bath and a microtubule extraction protocol. Polymerized microtubules were detected by immunofluorescence confocal microscopy and Western blot analyses. After cold exposure, immunofluorescence revealed that the majority of HeLa cell microtubules disassembled, whereas a smaller population of endothelial cell microtubules disassembled. Immunoblot analyses showed that microvascular endothelial cells express the microtubule cold-stabilizing protein N-STOP (neuronal stable tubule-only polypeptides), and that N-STOP binds to endothelial microtubules after cold exposure, but not if microtubules are disassembled with nocodazole before cold exposure. Hence, pulmonary endothelia have a population of cold-stable microtubules.

Original languageEnglish (US)
Pages (from-to)L132-L138
JournalAmerican Journal of Physiology - Lung Cellular and Molecular Physiology
Volume300
Issue number1
DOIs
StatePublished - Jan 1 2011
Externally publishedYes

Fingerprint

Microtubules
Endothelium
Lung
Population
Tubulin
Endothelial Cells
Population Dynamics
HeLa Cells
Nocodazole
Exotoxins
Peptides
Ice
Baths
Fluorescence Microscopy
Confocal Microscopy
Neuroglia
Pseudomonas aeruginosa
Fluorescent Antibody Technique
Cell Culture Techniques
Fibroblasts

Keywords

  • Endothelium
  • Stable tubule-only polypeptides

ASJC Scopus subject areas

  • Physiology
  • Pulmonary and Respiratory Medicine
  • Physiology (medical)
  • Cell Biology

Cite this

Cold exposure reveals two populations of microtubules in pulmonary endothelia. / Ochoa, Cristhiaan D.; Stevens, Troy; Balczon, Ron.

In: American Journal of Physiology - Lung Cellular and Molecular Physiology, Vol. 300, No. 1, 01.01.2011, p. L132-L138.

Research output: Contribution to journalArticle

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