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The Journal of Membrane Biology

, Volume 152, Issue 1, pp 13–23 | Cite as

Calcium-dependent Modulation of the Agonist Affinity of the Mammalian Olfactory Cyclic Nucleotide-gated Channel by Calmodulin and a Novel Endogenous Factor

  • S.  Balasubramanian
  • J.W.  Lynch
  • P.H.  Barry

Abstract.

The calcium-dependent modulation of the affinity of the cyclic nucleotide-gated (CNG) channels for adenosine 3′,5′-cyclic monophosphate (cAMP) was studied in enzymatically dissociated rat olfactory receptor neurons, by recording macroscopic cAMP-activated currents from inside-out patches excised from their dendritic knobs. Upon intracellular addition of 0.2 mm Ca2+ (0.2 Ca) the concentration of cAMP required for the activation of half-maximal current (EC50) was reversibly increased from 3 μm to about 30 μm. This Ca2+-induced affinity shift was insensitive to the calmodulin antagonist, mastoparan, was abolished irreversibly by a 2-min exposure to 3 mm Mg2++ 2 mm EGTA (Mg + EGTA), and was not restored by the application of calmodulin (CAM). Addition of CAM plus 0.2 mm Ca2+ (0.2 Ca + CAM), further reversibly shifted the cAMP affinity from 30 μm to about 200 μm. This affinity shift was not affected by Mg + EGTA exposure, but was reversed by mastoparan. Thus, the former Ca2+-only effect must be mediated by an unknown endogenous factor, distinct from CAM. Removal of this factor also increased the affinity of the channel for CAM. The affinity shift induced by Ca2+-only was maintained in the presence of the nonhydrolyzable cAMP analogue, 8-bromo-cAMP and the phosphatase inhibitor, microcystin-LR, ruling out modulation by phosphodiesterases or phosphatases. Our results indicate that the olfactory CNG channels are modulated by an as yet unidentified factor distinct from CAM.

Key words: Olfactory receptor neuron — CNG channel — Cyclic AMP — Ca2+— Calmodulin 

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Copyright information

© Springer-Verlag New York Inc. 1996

Authors and Affiliations

  • S.  Balasubramanian
    • 1
  • J.W.  Lynch
    • 2
  • P.H.  Barry
    • 1
  1. 1.School of Physiology and Pharmacology, University of New South Wales, Sydney 2052, AustraliaAU
  2. 2.Neurobiology Division, Garvan Institute of Medical Research, Darlinghurst, Sydney 2010, AustraliaAU

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