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Publication Detail
Calcium signalling mediated by the ?9 acetylcholine receptor in a cochlear cell line from the Immortomouse
  • Publication Type:
    Journal article
  • Publication Sub Type:
    Article
  • Authors:
    Jagger DJ, Griesinger CB, Rivolta MN, Holley MC, Ashmore JF
  • Publication date:
    2000
  • Pagination:
    49, 54
  • Journal:
    The Journal of Physiology
  • Volume:
    527
  • Print ISSN:
    0022-3751
  • Keywords:
    ACh, calcium, calcium signalling, cell, Cell Line, CELLS, cochlea, Fluorescence, hair cell, imaging, mouse, Permeability, physiology, Receptor, release, S, signalling, transgenic
  • Addresses:
    Department of Physiology, University College London, Gower Street, London WC1E 6BT, UK
Abstract
1. We have investigated the characteristics of the alpha9 acetylcholine receptor (alpha9AChR) expressed in hair cell precursors in an immortalized cell line UB/OC-2 developed from the organ of Corti of the transgenic H-2Kb-tsA58 mouse (the Immortomouse) using both calcium imaging and whole-cell recording. 2. Ratiometric measurements of fura-2 fluorescence revealed an increase of intracellular calcium concentration in cells when challenged with 10 microM ACh. The calcium increase was seen in 66 % of the cells grown at 39 degrees C in differentiated conditions. A sm aller fraction (34%) of cells grown at 33 degrees C in proliferative con ditions responded. 3. Caffeine (10mM) elevated cell calcium. In the ab sence of caffeine, the majority of imaged cells responded only once to A Ch presentations. Pretreatment with caffeine ingibited all calcium respo nses to ACh. 4. In whole-cell tight-seal recordings 10 microM ACh activa ted inward current was dependent on the extracellular calcium concentrat ion with an estimated PCa/PNa of 80 for the alpha9 receptor at physiological calcium levels. 5 . The data indicate that ACh activates a calcium-permeable channel alpha 9AChR in UB/OC-2 cells and that the channel has a significantly higher c alcium permeability than other AChRs. The results indicate that the alp ha9AChR may be able to elevate intracellular calcium levels in hair cell s both directly and via store release
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