Lenses
The wild type mouse strain used in this study was the 129SvEvTac mouse. Lenses examined were from 5(8 lenses), 46(8 lenses) and 72(6 lenses) wk old mice.
AlphaA/BKO was generated by cross breeding alphaAKO-127 [22] and alphaBKO-168 mice [23], also in a 129Sv background. These mice also lack the HSPB2 gene product [23]. Lenses examined were from 5 wk (6 lenses) and 54 wk (16 lenses) old mice. All animals were treated in accordance with the Association for Research in Vision and Ophthalmology Statement for the Use of Animals in Ophthalmic and Vision Research.
Immediately after euthanizing animals, eyes were enucleated. A scalpel blade was then used to make a small incision into the anterior chamber near the equator and then both eyes from individual animals were immersed in 5 ml fixative (2% (w/v) paraformaldehyde and 2% glutaraldehyde (v/v) in 0.1 M sodium cacodylate buffer, pH 7.2). Eyes were fixed for at least 24 hr at room temperature (RT) prior to dissection of the lenses. At this time, equatorial and axial dimensions of lenses and gross lenticular appearance were recorded. Statistical analyses on data consisted of Student's t-test, means and standard deviation, using the statistical software package StatMost (Dataxiom Software Inc., Los Angeles, CA).