Details
| Original language | English |
|---|---|
| Pages (from-to) | 85-94 |
| Number of pages | 10 |
| Journal | Journal of Bioenergetics and Biomembranes |
| Volume | 41 |
| Issue number | 1 |
| Publication status | Published - 4 Feb 2009 |
Abstract
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 IP 3-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 P2Y 2 and P2Y4 receptors in granulosa cells modulate Cl - permeability by regulating Ca2+-release.
Keywords
- Ca-imaging, Cl channels, Follicle maturation, Granulosa cells, Perforated patch-clamp, Purinergic receptors
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Physiology
- Biochemistry, Genetics and Molecular Biology(all)
- Cell Biology
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In: Journal of Bioenergetics and Biomembranes, Vol. 41, No. 1, 04.02.2009, p. 85-94.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Purinergic signalling in rat GFSHR-17 granulosa cells
T2 - An in vitro model of granulosa cells in maturing follicles
AU - Bintig, Willem
AU - Baumgart, Judith
AU - Walter, Wilhelm J.
AU - Heisterkamp, Alexander
AU - Lubatschowski, Holger
AU - Ngezahayo, Anaclet
N1 - Funding information: Acknowledgments The work was partly supported by the NANOTOME project; Biophotonik III.
PY - 2009/2/4
Y1 - 2009/2/4
N2 - 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 IP 3-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 P2Y 2 and P2Y4 receptors in granulosa cells modulate Cl - permeability by regulating Ca2+-release.
AB - 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 IP 3-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 P2Y 2 and P2Y4 receptors in granulosa cells modulate Cl - permeability by regulating Ca2+-release.
KW - Ca-imaging
KW - Cl channels
KW - Follicle maturation
KW - Granulosa cells
KW - Perforated patch-clamp
KW - Purinergic receptors
UR - http://www.scopus.com/inward/record.url?scp=63949086686&partnerID=8YFLogxK
U2 - 10.1007/s10863-009-9199-5
DO - 10.1007/s10863-009-9199-5
M3 - Article
C2 - 19191015
AN - SCOPUS:63949086686
VL - 41
SP - 85
EP - 94
JO - Journal of Bioenergetics and Biomembranes
JF - Journal of Bioenergetics and Biomembranes
SN - 0145-479X
IS - 1
ER -