Functional and ultrastructural changes in rabbit cornea during in vitro perfusion were studied by means of electrical measurements and electron microscopy. Electrical measurements were used to provide information on membrane potential, resistance, and capacitance. Membrane potential reflects cellular metabolic activity and thus is used to monitor tissue viability. On the other hand, since electrical resistance and capacitance indicate charge flow and the ability of the membrane dielectric to store charges, they can be used to detect structural changes in the aqueous transport pathway, i.e., intercellular space, and membrane surface, respectively. Studies on the current-voltage relationship of the cornea indicate self-generated membrane potential and non-ohmic resistive behavior of the cornea. However, the electrical resistance remains constant and is invariant to the external applied voltage. Problems associated with the use of ohmic resistance as an indication for membrane resistance are discussed. During in vitro perfusion, the cornea undergoes a biphasic change in resistivity with an initial rise and subsequent fall. The initial rise of the resistance appears to be induced by a change in membrane potential while the declining portion, at longer time periods, correlates well with the change in tissue morphology, i.e., expansion of the aqueous intercellular spaces. With the exception of this structural change, the overall integrity of the cornea is preserved and its viability is maintained for at least 6 h in vitro. The effect of polarization current on the d.c. electrical resistance was investigated by means of discharging kinetic measurements. Results indicate that under direct current, the cornea, due to its capacitive property, undergoes rapid, exponential-like, charging and discharging with an average time constant of approx. 50 μs. The polarization current, however, was found to have no significant effect on determination of membrane resistance. In addition to the kinetic study, the capacitive property of the cornea was further investigated by phase difference and impedance measurements. The use of a range of frequencies of measuring current results in a model of the cornea as electrically equivalent to a parallel RC circuit. Unlike resistance, the capacitance of the cornea remains remarkably constant during in vitro perfusion. This finding is in agreement with electron microscopic studies of the tissue. The use of electrical methods to study membrane transport properties as well as to assess tissue viability and integrity is discussed.
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