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Bharat Kumar, Wade Wesley Rich, Matthew Aaron Reilly; Changes in the localization of lens epithelial cell proliferation in response to stretching. Invest. Ophthalmol. Vis. Sci. 2020;61(7):4642.
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© ARVO (1962-2015); The Authors (2016-present)
The lens grows continuously throughout life and is driven by the proliferation of the lens epithelial cells (LECs). LEC proliferation has been observed to primarily occur near the lens equator in the germinative zone (GZ) in non-accommodating species. Recent work has shown that the proliferation of LECs is influenced by stretching the lens. This study hypothesizes that stretching the lens may alter the localization of LEC proliferation.
A porcine lens was dissected such that the ciliary body and a ring of sclera were still attached and mounted to a lens stretching device and stretched to a 12% change at the equatorial diameter. The lens was flat-mounted and stained for Ki-67, a proliferative marker, and with a general nuclear stain. A band of the lens capsule starting at the equator, passing through the anterior pole, and ending at the antipodal location was then imaged using a confocal microscope. A MATLAB script was used to determine the localization of proliferating cells on the lens capsule and quantify the localized Ki-67 labeling index as a function of distance from the anterior pole.
A mosaic of the confocal images show that Ki-67 labeling was primarily present near the equator, in the GZ (Figure 1). This was quantified by determining the localized labeling index of Ki-67 as a function of distance from the anterior pole by splitting the lens into ten regions. The percentage of the total number of cells that were labeled with Ki-67 in each region were quantified separately to determine the localized labeling index. The labeling index in the regions close to the anterior pole were < 1%, and the furthest regions in the GZ had a labeling index of 6.6% and 8.5% (Figure 2).
The proliferative activity in the statically stretched lens was mostly contained within the GZ. Future studies will examine if the distribution of Ki-67 labeling is altered in response to different stretching regimes, such as cyclic stretching and will further test the hypothesis that different stretching regimes may alter LEC proliferation localization.
This is a 2020 ARVO Annual Meeting abstract.
Figure 1. Confocal microscopy mosaic image of a band of statically stretched lens capsule labeled for Ki-67 (green) and with a general nuclear stain (gray) (A). The anterior pole (B). The GZ (C).
Figure 2. Localized Ki-67 labeling index as a function of distance from the anterior pole.
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