TY - JOUR

T1 - Regulation of ion fluxes, cell volume and gap junctional coupling by cGMP in GFSHR-17 granulosa cells

AU - Ngezahayo, A

AU - Altmann, B

AU - Kolb, H-A

PY - 2003/8/1

Y1 - 2003/8/1

N2 - Gap junctional communication between granulosa cells seems to play a crucial role for follicular growth and atresia. Application of the double whole-cell patch-clamp- and ratiometric fura-2-techniques allowed a simultaneous measurement of gap junctional conductance ( G(j)) and cytoplasmic concentration of free Ca(2+) ([Ca(2+)](i)) in a rat granulosa cell line GFSHR-17. The voltage-dependent gating of G(j) varied for different cell pairs. One population exhibited a bell-shape dependence of G(j) on transjunctional voltage, which was strikingly similar to that of Cx43/Cx43 homotypic gap junction channels expressed in pairs of oocytes of Xenopus laevis. Within 15-20 min, gap junctional uncoupling occurred spontaneously, which was preceded by a sustained increase of [Ca(2+)](i) and accompanied by shrinkage of cellular volume. These responses to the whole-cell configuration were avoided by absence of extracellular Ca(2+), blockage of K(+) efflux, or addition of 8-bromoguanosine 3',5'-cyclic monophosphate (8-Br-cGMP) to the pipette solution. Even in the absence of extracellular Ca(2+) or blockage of K(+) efflux, formation of whole-cell configuration generated a Ca(2+) spike that could be suppressed by the presence of 8-Br-cGMP. We propose that intracellular cGMP regulates Ca(2+) release from intracellular Ca(2+) stores, which activates sustained Ca(2+) influx, K(+) efflux and cellular shrinkage. We discuss whether gap junctional conductance is directly affected by cGMP or by cellular shrinkage and whether gap junctional coupling and/or cell shrinkage is involved in the regulation of apoptotic/necrotic processes in granulosa cells.

AB - Gap junctional communication between granulosa cells seems to play a crucial role for follicular growth and atresia. Application of the double whole-cell patch-clamp- and ratiometric fura-2-techniques allowed a simultaneous measurement of gap junctional conductance ( G(j)) and cytoplasmic concentration of free Ca(2+) ([Ca(2+)](i)) in a rat granulosa cell line GFSHR-17. The voltage-dependent gating of G(j) varied for different cell pairs. One population exhibited a bell-shape dependence of G(j) on transjunctional voltage, which was strikingly similar to that of Cx43/Cx43 homotypic gap junction channels expressed in pairs of oocytes of Xenopus laevis. Within 15-20 min, gap junctional uncoupling occurred spontaneously, which was preceded by a sustained increase of [Ca(2+)](i) and accompanied by shrinkage of cellular volume. These responses to the whole-cell configuration were avoided by absence of extracellular Ca(2+), blockage of K(+) efflux, or addition of 8-bromoguanosine 3',5'-cyclic monophosphate (8-Br-cGMP) to the pipette solution. Even in the absence of extracellular Ca(2+) or blockage of K(+) efflux, formation of whole-cell configuration generated a Ca(2+) spike that could be suppressed by the presence of 8-Br-cGMP. We propose that intracellular cGMP regulates Ca(2+) release from intracellular Ca(2+) stores, which activates sustained Ca(2+) influx, K(+) efflux and cellular shrinkage. We discuss whether gap junctional conductance is directly affected by cGMP or by cellular shrinkage and whether gap junctional coupling and/or cell shrinkage is involved in the regulation of apoptotic/necrotic processes in granulosa cells.

KW - Animals

KW - Calcium/metabolism

KW - Calcium Signaling/drug effects

KW - Cell Communication/drug effects

KW - Cell Size/drug effects

KW - Chelating Agents/pharmacology

KW - Connexins/metabolism

KW - Cyclic GMP/analogs & derivatives

KW - Cytoskeleton/drug effects

KW - Egtazic Acid/pharmacology

KW - Female

KW - Fura-2/metabolism

KW - Gap Junctions/drug effects

KW - Granulosa Cells/drug effects

KW - Ion Channel Gating

KW - Ion Transport/drug effects

KW - Patch-Clamp Techniques

KW - Potassium/metabolism

KW - Rats

U2 - 10.1007/s00232-003-2033-9

DO - 10.1007/s00232-003-2033-9

M3 - Article

C2 - 14502429

VL - 194

SP - 165

EP - 176

JO - The journal of membrane biology

JF - The journal of membrane biology

SN - 0022-2631

IS - 3

ER -