Carbon dioxide induced disassembly of gap-junctional plaques

When sheep ovarian follicle cells are maintained in an O2-rich environment their cells are metabolically coupled, as monitored by observing the exchange of [3H]choline; choline metabolites were detected up to 4 mm from the explant under these control conditions. When the tissues were placed in a CO2-rich environment the cells became uncoupled physiologically and choline metabolites were no longer exchanged. The cells in these two states, coupled and uncoupled, were examined by freeze-fracture. The initial controls were characteristic of ovarian follicular tissue exhibiting large macular plaques with regular outlines composed of PF intra-membranous particles (IMPs), which were arrayed in rows with IMP-free aisles. With uncoupling, the junctional plaques became irregular at the periphery, they became loosely packed and IMPs began to 'stream' out laterally across the membrane. Ultimately they were reduced to negligible IMP clusters or free IMPs. Analyses of the IMPs with an image analyser confirmed that in the uncoupled state the gap-junctional IMPs were dispersed over the membranes. On return to an O2-rich environment, the cells became recoupled as monitored by physiological criteria and in freeze-fracture replicas IMPs reclustered into macular, albeit smaller, plaques. These results support the contention that with uncoupling, gap-junctional particles are free to move and hence may become dispersed over the membrane face, with the possibility of being re-utilized to form junctions anew when conditions for coupling are re-established.