Differential protein expression in lens epithelial whole-mounts and lens epithelial cell cultures.
OBJECTIVE
Lens fibergenesis is a problem in several types of cataract and in the posterior capsular opacification following cataract surgery. To correct improper fiber differentiation or to prevent unwanted growth on the posterior capsule following cataract surgery requires a thorough understanding of normal and abnormal fiber formation. To this end, studies were initiated to characterize fiber differentiation in the bovine lens and in lens epithelial cell cultures.


METHODS
Indirect immunofluorescence and immunoblot analysis were employed to study the expression of vimentin, beta-crystallin, gamma-crystallin, filensin, aquaporin 0 and the Na, K-ATPase catalytic subunit isoforms (alpha1, alpha2, alpha3) in bovine lens epithelium whole-mounts as well as lens epithelial cell cultures propagated in medium containing 10% bovine serum or in medium supplemented with bovine serum concentrations < or =4%.


RESULTS
Three distinct cell types were observed in the bovine lens epithelium. The cells of the central zone were identified by a polarized distribution of two distinct Na, K-ATPase catalytic subunit isoforms, alpha1 to the apical (fiber side) and alpha3 to the basal (aqueous humor side) membranes. Lateral to the polarized central zone, was the germinative zone of cells, best characterized by perinuclear vimentin basket-like structures and the loss of polarized Na, K-ATPase catalytic subunit isoforms. Lateral to the germinative zone were the cells of the transition zone (meridinal rows) where expression of the lens specific proteins beta-crystallin, gamma-crystallin, filensin and aquaporin 0 as well as the lens fiber-, adipocyte- and brain glia-specific Na, K-ATPase catalytic subunit, alpha2 are expressed. The cultured cells propagated in medium supplemented with 10% serum bore no resemblance to any of the cells of the bovine lens epithelium whole-mounts. The cells propagated in the medium supplemented with the lower bovine serum levels resembled the differentiating fibers of the transition zone of the bovine lens epithelium whole-mounts as well as superficial cortical fibers.


CONCLUSIONS
Since the low-serum lens epithelial cell cultures bear a remarkable resemblance to early differentiating fibers, they are reasonable models for the study of early fiber differentiation or prevention of differentiation. The culture conditions employed do not yield the polarized cells of the central zone. Nor has the function of these polarized cells in lens fluid, nutrient and ion homeostasis been determined.
Ong
Marcia D
MD
Department of Pathology and Anatomy, Division of Cell Biology and Genetics, UNT Health Science Center, Fort Worth, TX 76107, USA.
Payne
D Michael
DM
Garner
Margaret H
MH
D000818:Animals; D020346:Aquaporins; D002386:Cataract / Q000473:pathology*; D002417:Cattle; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D003459:Crystallins / Q000032:analysis*; D003470:Culture Media; D046508:Culture Techniques; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism; D005136:Eye Proteins / Q000032:analysis; D019084:Fluorescent Antibody Technique, Indirect / Q000379:methods; D015151:Immunoblotting / Q000379:methods; D007381:Intermediate Filament Proteins / Q000032:analysis; D007527:Isoenzymes / Q000032:analysis; D007908:Lens, Crystalline / Q000166:cytology* / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000032:analysis; D012710:Serum Albumin, Bovine; D000254:Sodium-Potassium-Exchanging ATPase / Q000032:analysis; D014746:Vimentin / Q000032:analysis; D038204:beta-Crystallins / Q000032:analysis; D038222:gamma-Crystallins / Q000032:analysis
D020346:Aquaporins; D003459:Crystallins; D003470:Culture Media; D005136:Eye Proteins; D007381:Intermediate Filament Proteins; D007527:Isoenzymes; D008562:Membrane Glycoproteins; D014746:Vimentin; C030328:aquaporin 0; D038204:beta-Crystallins; C079098:filensin; D038222:gamma-Crystallins; D012710:Serum Albumin, Bovine; D000254:Sodium-Potassium-Exchanging ATPase