We study cell-to-cell channels, in cell pairs isolated from Chironomus salivary gland, by investigating the dependence of junctional conductance (gj) on membrane potentials (E1, E2), on Ca2+, and on H+, and we explore the interrelations among these dependencies; we use two separate voltage clamps to set the membrane potentials and to measure gj. We find gj to depend on membrane potentials whether or not a transjunctional potential is present. The pattern of gj dependence on membrane potentials suggests that each channel has two closure mechanisms (gates) in series. These gates pertain, respectively, to the two cell faces of the junction. By treating the steady-state gj as the resultant of two simultaneous but independent voltage-sensitive open/closed equilibria, one within each population of gates (i.e., one on either face of the junction), we develop a model to account for the steady-state gj vs. E relationship. Elevation of cytosolic Ca2+ or H+ at fixed E lowers gj, but at moderate concentrations of these ions this effect can be completely reversed by clamping to more negative E. Overall, the effect of a change in pCai or pHi takes the form of a parallel shift of the gj vs. E curve along the E axis, without change in slope. We conclude (1) that the patency of a cell-to-cell channel is determined by the states of patency of its two gates; (2) that the patency of the gates depends on membrane potentials (not on transjunctional potential), on pCai, and on pHi; (3) that pCai and pHi determine the position of the gj vs. E curve on the E axis; and (4) that neither Ca2+ nor H+ at moderate concentrations alters the voltage sensitivity of gj.

In four epithelial cell systems (salivary gland, renal, urinary bladder, and sensory cells) cells are interconnected as far as much of their ion content is concerned. In the salivary gland and renal epithelia, all cells of the epithelium are interconnected; and communication between a given cell and any of its nearest neighbors is equally good. In the bladder and sensory epithelia, communication appears to be more restricted, manifesting itself in chains of connected cells in the former, and in small groups of connected cells in the latter. The permeability of the cell membrane at the junction between connected cells is several orders of magnitude greater than it is at the cell surface bordering the exterior of the cells. Each connected cell ensemble functions as a system with a fairly continuous cytoplasmic core bounded by a diffusion barrier which is continuous along the entire outer surface of the system. As a result, ions move rather freely from cell to cell, but not from cell interior to exterior. Intercellular communication in at least three epithelia is associated with the presence of certain close-junctional membrane complexes.

The ion permeability of the membrane junctions between Chironomus salivary gland cells is strongly depressed by treatments that are generally known to inhibit energy metabolism. These treatments include prolonged coolhag at 6°-8°C, and exposure to dinitrophenol, cyanide, oligomycin, and Nethylmaleimide. IntraceUular injection of ATP appears to prevent depression of junctional permeability by dinitrophenol or to reverse it. Ouabain, azide, pchloromercuriphenylsulfonic acid, reserpine, and acetazolamide fail to depress junctional permeability. Thus the ion permeability of the junctional membranes appears to depend on energy provided by oxidative phosphorylation. Possible energy-linked processes for maintaining junctional permeability are discussed, including processes involving transport of permeability-modifying species such as Ca ++ .

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