In the calf eye lens, four morphologically distinct cell types can be detected: three in the epithelial monolayer and one in the cortical part. During differentiation, there is a quantitative change in the synthesis of crystallin subunits. A marked increase in aA-chains and several fl-crystallin polypeptides accompanies the transition from epithelial to fiberlike lens cells while synthesis of the noncrystallin proteins diminishes significantly.
KEY WORDS lens differentiation protein biosynthesis crystallinsThe lens of the mammalian eye has been explored for study of the typical structural proteins, the crystaUins (15,17,21,16,12,1), and recently for the investigation of membrane proteins and their biosynthesis (4,22,19,20). The protein biosynthetic events in the lens cortex which contains the bulk of the terminally differentiated fiber cells has been studied intensively (6). However, the question as to which crystallin polypeptide chains are synthesized in the epithelial monolayer has not been answered unequivocally. It would be extremely important to have the answer to this question in order to fruitfully investigate fundamental processes such as regulatory mechanisms in differentiation. Studies have been undertaken to establish the protein pattern of lens epithelia. Papaconstantinou (14) and Van Venrooij et al. (18) claimed the presence of c~-and fl-crystali~ns in the lens epithelium; on the other hand, according to Papaconstantinou, v-crystallin could only be found in lens fibers. The latter finding should, therefore, reflect an aspect of cellular differentiation in the lens. Biosynthesis of a-crystallin in the epithelium has been reported by Delcour and Papaconstantinou (9). In our opinion, in all investigations undertaken so far, contamination of the epithelial preparations with fibers could not be excluded. This is due to the fact that epithelial cells were defined as those cells that adhered to the lens capsule when it was removed from the organ. However, no evidence was provided that fiber cells were completely removed.In this paper, we describe a study of the synthesis of crystallins in the eye lens epithelium. Microscope observation revealed the absence of fiber cells in our preparations. Evidence is provided that the Bz chain of a-crystallin is preferentially synthesized in a defined part of the lens epithelium.
MATERIALS AND METHODS
L-[asS]Methionine (sp act 180 Ci/p.mol was purchased from the Radiochemical Centre (Amersham, England).
Tissue PreparationCalf eyes were obtained on ice from the slaughterhouse. They were washed w!th distilled water and opened at the lateral side, so that the lenses could be removed without adhering iris material. For microscope examination, the epithelia were spread on a microscope slide, as described in Results. The preparations were then fixed in OsO4 vapor for 3 min and stained according to Giemsa for 30 min (11).