Protocols to Isolate and Characterize Ciliary Muscle Cells and Schlemm's Canal Cells and Detect Matrix Metalloproteases

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Ocular hypertension and the ensuing glaucomatous optic neuropathy occur when the aqueous humor drainage system within the anterior chamber of the eye, trabecular meshwork (TM), and its associated Schlemm's canal (SC) system become clogged due to the abnormally deposited excess extracellular matrix components. A similar series of events plague the ciliary muscle (CM) bundles in the uveoscleral pathway for AQH drainage. In order to study pathological events and endogenous chemicals associated with the latter tissues, isolated cells of TM, SC, and CM have proven useful. This chapter provides a brief and simple description of protocols to 1) isolate and characterize ciliary smooth muscle cells, 2) isolate and characterize Schlemm's canal cells, and 3) measure enzyme activities and expression pattern of matrix metalloproteases (MMPs) secreted from the isolated ocular cells using zymography and Western blotting. These studies are important for the characterization of the biochemical pharmacology of the drugs that activate specific receptors and enzymes that mediate the aqueous humor outflow from the anterior chamber of the eye to lower the IOP.