Purpose: To elucidate the mechanical properties of the bovine lens cortical membrane (CM), the nuclear membrane (NM) containing cholesterol bilayer domains (CBDs), and whole bovine lenses.
Methods: The total lipids (lipids plus cholesterol) from the cortex and nucleus of a single bovine lens were isolated using the monophasic methanol extraction method. Supported CMs and NMs were prepared from total lipids extracted from the cortex and nucleus, respectively, using a rapid solvent exchange method and probe-tip sonication, followed by the fusion of unilamellar vesicles on a flat, freshly cleaved mica surface. Topographical images and force curves for the CMs and NMs were obtained via atomic force microscopy (AFM) in a fluid cell. Whole bovine lenses were affixed to custom-built glass Petri dishes, and an AFM was used to obtain force curves. Force curves were analyzed to estimate the breakthrough force, membrane stiffness (KA and Em), and lens stiffness (EL).
Results: The NMs containing CBDs exhibited significantly lower breakthrough force, KA, and Em than the CMs without CBDs. The Em values for CMs and NMs were significantly higher than the EL for the whole lens.
Conclusions: The significantly higher stiffness of the CM and NM compared to the stiffness of the whole lens suggests that slight modulation in CM and NM composition may play a crucial role in altering the overall lens stiffness. Furthermore, the NMs containing CBDs were less stiff than CMs without CBDs, suggesting that CBDs decrease lens membrane stiffness and possibly protect against lens hardening and presbyopia.