Prostaglandin F2 alpha induces unsynchronized intracellular calcium oscillations in monolayers of gap junctionally coupled NRK fibroblasts

Abstract

We investigated the intracellular calcium oscillations induced by prostaglandin F2alpha (PGF2alpha) in individual cells of confluent, gap junction-coupled monolayers of normal rat kidney (NRK) fibroblasts. PGF2alpha (1000 nM) induced oscillations in more than 90% of the cells in the monolayer, but the frequency of these oscillations was highly variable between individual cells (0.2-1.4 min(-1)). The initial calcium peak resulted from calcium release from IP3-sensitive stores, while subsequent calcium transients were mediated by interplay between both IP3-sensitive calcium stores and calcium influx. The oscillation frequency was increased by sensitizing the IP3 receptor with thimerosal (10 microM) and depended on the extracellular calcium concentration. Thapsigargin (5 nM), which inhibits reuptake of calcium into the stores, only seemed to reduce the amplitude of the oscillation. Patch-clamp experiments revealed that PGF2alpha did not inhibit electrical coupling of the NRK cells in the monolayer. Gap junctional permeability of NRK cells thus appears to be sufficient to allow electrical coupling, resulting in a uniform membrane potential throughout the entire monolayer, but insufficient to synchronize the intracellular calcium oscillations upon PGF2alpha stimulation.