Chronic Fluid Flow Is an Environmental Modifier of Renal Epithelial Function
Figure 9
The purpose of the overall experiment was to probe the ciliary-mediated mechanosensation response to chronic fluid flow. Monolayers were also exposed to specific changes in flow rates to assess the dynamics of the mechanosensation response. Previous experiments have shown that the transepithelial sodium current is a valid readout that requires several days to reach steady-state conditions. Thus, monolayers were maintained in constant conditions for 10 day blocks of time. Finally, by choosing different start times relative to the onset of differentiation, the role of initial flow conditions as a modifier of future function could be assessed. In order to uniquely identify each group of monolayers for analysis, a three-digit alphanumeric code was assigned based on the particular pattern of applied flow conditions and start time relative to the onset of differentiation. Cohorts 1 and 2 started the protocol 72 hours prior to entering differentiation conditions, cohorts 3 and 4 began the protocol at the onset of differentiation conditions, and cohorts 5 and 6 began the flow protocol 10 days after the onset of differentiation conditions.