Protocols for Studying Aquaporin Function Related to Ocular Hypertension In Vitro

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Aquaporins (AQPs) are a large family of membrane-bound water channels that facilitate water and small solute transport and have important physiological functions in all cells. At least thirteen types of aquaporins exist that exhibit a high level of phylogenetic homology, and many are present in ocular cells. Mouse knockout studies have demonstrated that AQPs play an important function in aqueous humor (AQH) dynamics in the anterior chamber cells and also have a role in retinal function. In contrast to the rich literature on aquaporin function, very little is known about how to assess water movement mediated by AQPs in their native environment, such as across ciliary epithelial cell membranes where AQH is manufactured. In part, the reason could be the lack of appropriate procedures for studying water movement across ciliary epithelial layers. This chapter describes methods to measure AQP function in vitro using ciliary epithelial cells, including a high-throughput screening assay method. In addition, a procedure to determine aquaporin function in its native environment is also presented.