Finger-like projections of plasma membrane in the most senescent fiber cells of human lenses.
OBJECTIVE
To corroborate the findings of finger-like membrane projections in monkey and baboon lenses, in human lens fiber cells.


METHODS
Normal human lenses, two months to 76 years old, as well as age-related nuclear cataracts, were immersion fixed in 2% paraformaldehyde-0.2% glutaraldehyde in 0.1 M sodium cacodylate buffer pH 7.2 for 24 h at room temperature, cut into 200-500 microm thick sections along the equatorial axis, fixed for an additional 12-18 h at room temperature, dehydrated in an ascending ethanol series and embedded in Lowicryl K4M. Semi-thick sections (0.25-1.0 microm) were cut, absorbed onto 75 or 100 nickel slotted grids, labeled with anti-MIP 26 or phalloidin, stained with 2% uranyl acetate and viewed by transmission electron microscopy at 100 kV.


RESULTS
Transmission electron microscopy micrographs demonstrated the presence of finger-like plasma membrane projections measuring 0.16-0.25 microm in diameter and 1.0-6.5 microm in length with bulbous terminal tips in the most senescent fiber cells in two-month and older normal lenses, as well as, in nuclear cataracts. These projections appeared to overlie furrowed membrane domains in the extracellular space, as well as project into the cytosol along the cytosolic leaflet of plasma membrane.


CONCLUSIONS
The results extend the findings in monkey and baboon lenses, to the human lens, and demonstrate that these projections, which sparsely label with antiserum against MIP 26, but not filamentous actin, not only extend into the extracellular space, but also project inward into the cytosol.
Boyle
D L
DL
Division of Biology, Kansas State University, Manhattan 66506, USA. dboyle@ksu.edu
Takemoto
L J
LJ
D000199:Actins / Q000032:analysis; D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000375:Aging; D000818:Animals; D020346:Aquaporins; D002386:Cataract / Q000473:pathology; D002462:Cell Membrane / Q000737:chemistry / Q000648:ultrastructure; D016922:Cellular Senescence; D002648:Child; D005136:Eye Proteins / Q000032:analysis; D006801:Humans; D007223:Infant; D007908:Lens, Crystalline / Q000737:chemistry / Q000648:ultrastructure*; D008251:Macaca; D008562:Membrane Glycoproteins / Q000032:analysis; D016253:Microscopy, Immunoelectron; D008875:Middle Aged; D010215:Papio
D000199:Actins; D020346:Aquaporins; D005136:Eye Proteins; D008562:Membrane Glycoproteins; C030328:aquaporin 0