Calcium localization and ultrastructure of clear and pCMPS-treated rat lenses

The sulfhydryl complexing agent p-chloromercuri-phenylsulfonate (pCMPS) has been shown to increase lens membrane permeability, Na+ and Ca2+ content, and light scatter in the rat lens in vitro. This study aimed to investigate the ultrastructural changes accompanying the increase in light scatter. In addition, high-resolution histochemistry was used to study the cellular distribution of Ca2+ in normal and cataractous lenses. Rat lenses were incubated for 4 hours in normal (1 mM) and high (5 mM) Ca2+ containing media supplemented with 40 microM pCMPS. Control lenses were incubated in 1 mM Ca2+ containing medium. They were prepared for scanning and transmission electron microscopy and for Ca2+ localization using the oxalate-pyroantimonate procedure. Control lenses incubated for 4 hours had normal morphology and showed no evidence of light scatter. Calcium distribution as observed with the oxalate-pyroantimonate precipitation method was low in superficial fibers, high in the membranes of intermediate fibers, and declined again toward the nucleus. In the deeper cortex, there also were small vacuoles of calcium accumulation. pCMPS treatment (in 1 and 5 mM Ca2+) induced a significant influx of calcium into the lens cytoplasm. Calcium-containing extracellular vacuoles also were seen in the intermediate cortex in both cases. The presence of these vacuoles appeared to correlate with the major areas of light scatter in the lens. In 5 mM Ca2+, intracellular vacuoles were observed throughout the superficial cortex. Most of the calcium observed by oxalate-pyroantimonate in the normal lens is located at the membrane, and the staining appears strongest in the intermediate cortex. In pCMPS treatment, large extracellular vacuoles are present in this intermediate zone and appear to be the major source of light scatter. This zone may be the initiation site of many different types of cataract, including some described in human lenses.