Effects of methyl-β-cyclodextrin treatment on secretion profile of interferon-/? and zonula occuludin-1 architecture in madin-darby canine kidney cell monolayers

Masato Maruyama, Kayo Ishida, Yoshihiko Watanabe, Makiya Nishikawa, Yoshinobu Takakura

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

The interferon (IFN) is a paradigm of secretory protein. However, it has been poorly understood how its secretion is regulated in polarized epithelial cells. Recently, we had shown that exogenous IFNs transiently expressed in polarized monolayers were predominantly secreted to the side on which gene transfection had been performed, while stably expressed IFNs were secreted almost equally to the both cell sides. Since those modes of secretion did not affect each other, epithelial cell layers seemed to have at least two protein sorting/secretion pathways, one for transient expression and the other for stable expression, for identical secretory proteins. Furthermore, this dual secretion profile seemed to be mediated by distinct post-trans Golgi network vesicles, suggesting the involvement of lipid rafts in the sorting multiplicity. To address this issue, here we studied the effects of cholesterol depletion with methyl-//-cyclodextrin (M/CD) on the secretion profile of IFN-/ exogenously expressed in Madin-Darby canine kidney (MDCK) cells. The M/CD-treatment, however, did not affect the profile in either transient or stable expression, although the architecture of zonula occuludin-1, which links to the tight junction, was substantially disrupted by the treatment. Further analysis of Triton X-100-insoluble cell extracts by sucrose density centrifugation demonstrated that IFN-/ was not apparently associated with lipid rafts in either transient or stable expression. These results suggest that lipid rafts may not be crucially involved in the regulation of secretion polarity of IFN-/ in the epithelial cells.

Original languageEnglish
Pages (from-to)910-915
Number of pages6
JournalBiological and Pharmaceutical Bulletin
Volume32
Issue number5
DOIs
Publication statusPublished - May 1 2009
Externally publishedYes

Fingerprint

Madin Darby Canine Kidney Cells
Cyclodextrins
Interferons
Epithelial Cells
Lipids
trans-Golgi Network
Tight Junctions
Secretory Pathway
Octoxynol
Protein Transport
Cell Extracts
Centrifugation
Transfection
Sucrose
Proteins
Cholesterol
Genes

Keywords

  • Epithelial cell
  • Interferon-/
  • Lipid raft
  • Methyl-/j-cyclodextrin
  • Secretion polarity
  • Zonula occludin-1

ASJC Scopus subject areas

  • Pharmacology
  • Pharmaceutical Science

Cite this

Effects of methyl-β-cyclodextrin treatment on secretion profile of interferon-/? and zonula occuludin-1 architecture in madin-darby canine kidney cell monolayers. / Maruyama, Masato; Ishida, Kayo; Watanabe, Yoshihiko; Nishikawa, Makiya; Takakura, Yoshinobu.

In: Biological and Pharmaceutical Bulletin, Vol. 32, No. 5, 01.05.2009, p. 910-915.

Research output: Contribution to journalArticle

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