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Inactivation of the murine cftr gene abolishes cAMP-mediated but not Ca2+-mediated secretagogue-induced volume decrease in small-intestinal crypts

Abstract

The cellular volume of crypts isolated from 2- to 3-week-old mouse small intestine has been measured to assess the capacity of the epithelial cells to respond to secretagogues. Vasoactive intestinal polypeptide (VIP) or carbachol, respectively cAMP- and calcium-mediated secretagogues, produced a reduction crypt volume attributed to KCl loss through channels activated by the agonists. Consistent with the participation of separate chloride channels, 4,4′-diisothiocyanatostilbene-2,2′-disulphonic acid (DIDS) blocked the carbacholbut not the VIP-induced volume decrease, whilst gl- ibenclamide abolished the VIP effect without affecting the carbachol-induced volume decrease. Animals homozygous for a disrupted cftr gene, introduced by gene targeting, were also used as the source for crypt isolation. In these CFTR(-/-) crypts, VIP failed to elicit any reduction in cellular volume, while the response to carbachol was indistinguishable from that seen in crypts from age-matched control animals. These results are consistent with murine CFTR being a cAMP-activated chloride channel inhibited by glibenclamide and resistant to DIDS. A separate chloride conductance activated by calcium mobilization in small-intestinal crypts appears to be independent of CFTR.

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Correspondence to M. A. Valverde.

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Valverde, M.A., O'Brien, J.A., Sepúlveda, F.V. et al. Inactivation of the murine cftr gene abolishes cAMP-mediated but not Ca2+-mediated secretagogue-induced volume decrease in small-intestinal crypts. Pflugers Arch. 425, 434–438 (1993). https://doi.org/10.1007/BF00374869

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Key words

  • Small-intestinal crypts
  • Intestinal secretion
  • Cystic fibrosis
  • Chloride conductance
  • DIDS
  • Glibenclamide
  • Mouse