当前位置： SCI文献检索 > INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE期刊下所有文献 > Confocal microscopy reveals zones of membrane remodeling in the outer cortex of the human lens.

Confocal microscopy reveals zones of membrane remodeling in the outer cortex of the human lens.

Abstract：

PURPOSE:To optimize fixation, sectioning, and immunolabeling protocols to map the morphology of the human lens with confocal microscopy. METHODS:Transparent human lenses were fixed in 0.75% paraformaldehyde for 24 hours, cut in half, and fixed for another 24 hours. Lenses were cryoprotected, sectioned, and labeled with wheat germ agglutinin, aquaporin-0 antibodies, Hoechst, or toluidine blue. Before fixation, some lenses were incubated in an extracellular marker dye, Texas Red-dextran. Labeled sections were imaged with a confocal microscope. Overlapping images were tiled together to form a continuous image montage of fiber cell morphology from the periphery to the lens center. RESULTS:Fiber cell morphologies were identical with those previously described by electron microscopy and allowed immunohistochemistry to be performed for a representative membrane protein, aquaporin-0. Sectioning protocols enabled the epithelium and outer cortex to be retained, leading to the identification of two unique morphologic zones. In the first zone, an age-independent compaction of nucleated fiber cells and the initiation of extensive membrane remodeling occur. In the second zone, fiber cells retain their interdigitations but lose their nuclei, exhibit a distorted shape, and are less compressed. These zones are followed by the adult nucleus, which is marked by extensive compaction and a restriction of the extracellular space to the diffusion of Texas Red-dextran. CONCLUSIONS:The authors have developed sectioning and imaging protocols to capture differentiation-dependent changes in fiber cell morphology and protein expression throughout the human lens. Results reveal that differentiating fiber cells undergo extensive membrane remodeling before their internalization into the adult nucleus.

journal_name

Invest Ophthalmol Vis Sci

journal_title

Investigative ophthalmology & visual science

authors

Lim JC,Walker KL,Sherwin T,Schey KL,Donaldson PJ

doi

10.1167/iovs.09-3435

subject

Has Abstract

pub_date

2009-09-01 00:00:00

pages

4304-10

issue

eissn

0146-0404

issn

1552-5783

pii

iovs.09-3435

journal_volume

pub_type

杂志文章

在线工具

Confocal microscopy reveals zones of membrane remodeling in the outer cortex of the human lens.