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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

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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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Authors and Affiliations

  1. AFRC Institute of Animal Physiology and Genetics Research, Cambridge Research Station, CB2 4AT, Cambridge, UK

    M. A. Valverde, J. A. O'Brien & F. V. Sepúlveda

  2. Wellcome/CRC Institute of Cancer and Developmental Biology and Department of Genetics, University of Cambridge, Tennis Court Road, CB2 1QR, Cambridge, UK

    R. Ratcliff, M. J. Evans & W. H. Colledge

Authors
  1. M. A. Valverde
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  2. J. A. O'Brien
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  3. F. V. Sepúlveda
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  4. R. Ratcliff
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  5. M. J. Evans
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  6. W. H. Colledge
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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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  • DOI: https://doi.org/10.1007/BF00374869

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