Intra- and transepithelial analytical techniques

Epithelia transport a variety of solutes and water. Study of such transport requires a determination of the driving forces responsible for transport, of the pathways through which transport occurs, and of the factors controlling such transport. Transepithelial driving forces are readily determined where the composition of the bathing media can be altered and electrical forces negated. Where substances move only through a paracellular pathway such manipulations may be adequate to define the permeability and selectivity of the pathways. For substances utilizing a cellular pathway, driving forces and permeabilities across the two dissimilar apical and basolateral cellular membranes must be determined. Where a substance can be shown to move across a membrane against its electrochemical potential gradient, the source of the energy for such movement must be assessed. This review focuses on the applicability and validity of a variety of techniques utilized for the study of epithelial transport to answer these questions. These include microelectrode techniques, chemical analyses, microprobe analysis, microscopy, and techniques for assessing the coupling of metabolism to transport.