Immunocytochemical evidence for an actin-myosin system in lens epithelial cells.
Since filamentous actin had been shown earlier to exist in lens epithelial and fiber cells, we inquired whether this could represent a contractile system with myosin and other actin-associated proteins. We resolved this question in freshly removed or organ-cultured rabbit and squirrel lens epithelial whole mounts using immunocytochemical techniques and by immunoblots of extracts separated by electrophoresis. In the former, methods were developed using long fixation times and long incubation in primary antibodies and biotinylated second antibodies visualized by streptavidin immunofluorescence and by diaminobenzidine peroxidase. Myosin was found to be localized along the filamentous rays and at central vertices of polygonal arrays situated at the apices of epithelial cells. It was not clear whether myosin and actin occurred together along the same or adjacent filaments in a bundle. Tubulin and vimentin were found deeper in the cells and were not aligned with actin and myosin filaments. Control lens epithelia treated similarly except for deletion of the primary antibodies showed no staining. As positive controls, pieces of glycerinated sartorius muscle exhibited characteristic cross-banded patterns of actin and myosin when incubated with the same reagents used on the lens epithelium. Denatured extracts of rabbit lens epithelium and of cortical fiber cells separated by electrophoresis and transferred to nitrocellulose paper, stained specifically with the same myosin and tubulin antibodies used in the immunocytochemistry experiments. The molecular weight profile of the myosin polypeptide indicated that lens tissue has myosin II. We conclude that a contractile system exists in lens epithelial and cortical fiber cells, although the function is not understood at this time. We conjecture that the system may act to stabilize lens shape by providing contractile tone.
Rafferty
N S
NS
Department of Cell, Molecular and Structural Biology, Northwestern University, Chicago, IL 60611.
Scholz
D L
DL
Goldberg
M
M
Lewyckyj
M
M
D000199:Actins / Q000032:analysis*; D000818:Animals; D004848:Epithelium / Q000737:chemistry; D007150:Immunohistochemistry; D007908:Lens, Crystalline / Q000737:chemistry*; D009218:Myosins / Q000032:analysis*; D009924:Organ Culture Techniques; D011817:Rabbits; D012589:Sciuridae; D014404:Tubulin / Q000032:analysis; D014746:Vimentin / Q000032:analysis
D000199:Actins; D014404:Tubulin; D014746:Vimentin; D009218:Myosins