Effect of pneumolysin on rat brain ciliary function: comparison of brain slices with cultured ependymal cells

R A Hirst, A Rutman, K Sikand, P W Andrew, T J Mitchell, C O'Callaghan

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28 Citations (Scopus)

Abstract

This study compares two models for examining ependymal ciliary function: rat brain slices cut from the fourth ventricle and primary ependymal cells in culture. The cilia from both preparations were very reproducible; each preparation had cilia beating at a constant frequency of between 38 and 44 Hz. With the brain slices, ciliary stasis occurred after 5 d in culture. However, ependymal cells had fully functional cilia for up to 48 d in culture. The pneumococcal toxin, pneumolysin, caused a dose-dependent inhibition of cilia beat frequency within 15 min in both models. There were no significant differences in the mean log 50% inhibitory concentration (pIC50) slice = 0.65 +/- 0.05, equivalent to 4.4 hemolytic units (HU)/mL; cells = 0.57 +/- 0.14, equivalent to 3.7 HU/mL. There were also no significant differences in the mean Hill slope factors for the curves (slice = 1.4 +/- 0.05; cells = 1.6 +/- 0.4). These data demonstrate that both models can be used to examine the acute (15-min) effects of pneumolysin on cilia beat frequency. The main advantage of the primary ependymal culture model is that considerably more cultured ependymal cells (approximately 70%) are available, compared with the number of ependymal cells on the brain slices (approximately 2%), thus reducing the number of animals used. A pure ependymal culture was not achieved (approximately 30% of the cells were not ciliated). The increased survival time of the ependymal cells compared with the brain slices make cultured ependymal cells more useful for examining long-term ciliary function, whereas brain slices may be more useful for examining the interactions between ependymal and other nearby cells.

Original languageEnglish
Pages (from-to)381-4
Number of pages4
JournalPediatric Research
Volume47
Issue number3
Publication statusPublished - Mar 2000

Keywords

  • Animals
  • Bacterial Proteins
  • Cells, Cultured
  • Cerebellum
  • Cilia
  • Ependyma
  • In Vitro Techniques
  • Microscopy, Electron, Scanning
  • Rats
  • Rats, Wistar
  • Streptolysins

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