Figure 5. Effects of NCC and NKCC inhibitors on lens transparency and fiber cell morphology. Rat lenses were organ cultured for 18 h in isotonic AAH in either the absence (A) or presence of the NCC inhibitor thiazide (B) or the NKCC inhibitor bumetanide (C). Lens transparency was monitored by dark field microscopy while fiber cell morphology was determined by imaging equatorial sections labeled with the membrane marker WGA. Culturing lenses in AAH only (A) or AAH + 10 μM thiazide (B) had no major effects on lens transparency (left panels) or fiber cell morphology (right panels). C: Culturing lenses in AAH + 2 μM bumetanide caused cortical opacification of the lens (left panel) that was induced by damage to cortical fiber cell morphology (right panel). D-F: High powered images from the areas indicated (boxes) in A and C. Fiber cells in the lens periphery exhibited a marked shrinkage in the presence of bumetanide (D) relative to that observed in the absence of the inhibitor (E), while in the deeper influx zone dilations of the extracellular space between fiber cells (F) was observed in the presence of bumetanide.