Plasma membrane Ca2+-ATPase in the cilia of olfactory receptor neurons: possible role in Ca2+ clearance

Authors

  • Karen Castillo,

    1. Department of Biology, Faculty of Sciences and Millennium Institute for Cell Dynamics and Biotechnology, University of Chile, Santiago, Chile
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  • Ricardo Delgado,

    1. Department of Biology, Faculty of Sciences and Millennium Institute for Cell Dynamics and Biotechnology, University of Chile, Santiago, Chile
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  • Juan Bacigalupo

    1. Department of Biology, Faculty of Sciences and Millennium Institute for Cell Dynamics and Biotechnology, University of Chile, Santiago, Chile
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Dr J. Bacigalupo, as above.
E-mail: bacigalu@uchile.cl

Abstract

Olfactory sensory neurons respond to odorants increasing Ca2+ concentrations in their chemosensory cilia. Calcium enters the cilia through cAMP-gated channels, activating Ca2+-dependent chloride or potassium channels. Calcium also has a fundamental role in odour adaptation, regulating cAMP turnover rate and the affinity of the cyclic nucleotide-gated channels for cAMP. It has been shown that a Na+/Ca2+ exchanger (NCX) extrudes Ca2+ from the cilia. Here we confirm previous evidence that olfactory cilia also express plasma membrane Ca2+-ATPase (PMCA), and show the first evidence supporting a role in Ca2+ removal. Both transporters were detected by immunoblot of purified olfactory cilia membranes. The pump was also revealed by immunocytochemistry and immunohistochemistry. Inside-out cilia membrane vesicles transported Ca2+ in an ATP-dependent fashion. PMCA activity was potentiated by luminal Ca2+ (K0.5 = 670 nm) and enhanced by calmodulin (CaM; K0.5 = 31 nm). Both carboxyeosin (CE) and calmidazolium reduced Ca2+ transport, as expected for a CaM-modulated PMCA. The relaxation time constant (τ) of the Ca2+-dependent Cl current (272 ± 78 ms), indicative of luminal Ca2+ decline, was increased by CE (2181 ± 437 ms), by omitting ATP (666 ± 49 ms) and by raising pH (725 ± 65 ms), suggesting a role of the pump on Ca2+ clearance. Replacement of external Na+ by Li+ had a similar effect (τ = 442 ± 8 ms), confirming the NCX involvement in Ca2+ extrusion. The evidence suggests that both Ca2+ transporters contribute to re-establish resting Ca2+ levels in the cilia following olfactory responses.

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