Purinergic signalling in rat GFSHR-17 granulosa cells: an in vitro model of granulosa cells in maturing follicles

  • Willem Bintig
  • Judith Baumgart
  • Wilhelm J. Walter
  • Alexander Heisterkamp
  • Holger Lubatschowski
  • Anaclet NgezahayoEmail author


Purinergic signalling in rat GFSHR-17 granulosa cells was characterised by Ca2+-imaging and perforated patch-clamp. We observed a resting intracellular Ca2+-concentration ([Ca2+]i) of 100 nM and a membrane potential of −40 mV. This was consistent with high K+− and Cl permeability and a high intracellular Cl concentration of 40 mM. Application of ATP for 5–15 s every 3 min induced repeated [Ca2+]i increases and a 30 mV hyperpolarization. The phospholipase C inhibitor U73122 or the IP3-receptor antagonist 2-aminoethoethyl diphenyl borate suppressed ATP responses. Further biochemical and pharmacological experiments revealed that ATP responses were related to stimulation of P2Y2 and P2Y4 receptors and that the [Ca2+]i increase was a prerequisite for hyperpolarization. Inhibitors of Ca2+-activated channels or K+ channels did not affect the ATP-evoked responses. Conversely, inhibitors of Cl channels hyperpolarized cells to −70 mV and suppressed further ATP-evoked hyperpolarization. We propose that P2Y2 and P2Y4 receptors in granulosa cells modulate Cl permeability by regulating Ca2+-release.


Granulosa cells Purinergic receptors Cl channels Follicle maturation Perforated patch-clamp Ca2+-imaging 


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Copyright information

© Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Willem Bintig
    • 1
  • Judith Baumgart
    • 2
  • Wilhelm J. Walter
    • 3
  • Alexander Heisterkamp
    • 2
  • Holger Lubatschowski
    • 2
  • Anaclet Ngezahayo
    • 1
    Email author
  1. 1.Institute of BiophysicsLeibniz University HannoverHannoverGermany
  2. 2.Laser Zentrum Hannover e.V.HannoverGermany
  3. 3.Molecular and Cell PhysiologyHannover Medical SchoolHannoverGermany

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