Actin Filament Bundles in Cortical Fiber Cells of the Rat Lens

The distribution and organization of actin filament bundles were studied in cortical fiber cells of rat lenses at various ages (3 days to 2.5 months old), using thin-section electron microscopy, immunocytochemistry and immunoblotting. Electron microscopy showed that actin bundles were regularly found along cortical fiber cell membranes of the lens at all ages studied. The actin bundles were commonly arranged in three distinct units, one bundle in each fiber cell, located at the intersections where three hexagonal fiber cells meet as seen in cross sections. These actin bundles were approximately 150 nm in diameter and were composed of 7-nm small filaments. They were aligned parallel to the long axis of fiber cells as judged by both cross and longitudinal sections. The outside border of each bundle was always surrounded by a zone of 10-nm intermediate filaments which have the same orientation as that of the actin bundles. In longitudinal sections, elongated actin bundles were always parallel to the cell membranes. A number of individual actin bundles sometimes were found to form a chain with periodic short intervals. In addition, actin bundles were frequently associated with adherens junctions near the intersections and other regions of fiber cell membranes. By immunoelectron microscopy, we demonstrated that these filament bundles indeed contained actins. By rhodamine-phalloidin labelling, we found that labeled actin bundles appeared as large, distinct dots at the corners of hexagonal fiber cells in all ages studied. In addition, non-bundle F-actins were labeled preferentially along the cell membranes of the short sides of hexagonal fiber cells. This resulted in a unique zigzag pattern of actin labeling commonly seen in the cortical fiber cells of a mature rat lens. Finally, we showed that alpha-actinin was associated with the actin bundles in the fiber cells by immunofluorescent double labeling and immunoblotting. It is suggested that this unique arrangement of actin bundles in fiber cells may provide a stabilizing structure for forming a sharp angle at each corner of fiber cells, thereby the hexagonal shape of the cells can be maintained.