Eph-ephrin Signaling Affects Eye Lens Fiber Cell Intracellular Voltage and Membrane Conductance

The avascular eye lens generates its own microcirculation that is required for maintaining lifelong lens transparency. The microcirculation relies on sodium ion flux, an extensive network of gap junction (GJ) plaques between lens fiber cells and transmembrane water channels. Disruption of connexin proteins, the building blocks of GJs, or aquaporins, which make up water and adhesion channels, lead to lens opacification or cataracts. Recent studies have revealed that disruption of Eph-ephrin signaling, in particular the receptor EphA2 and the ligand ephrin-A5, in humans and mice lead to congenital and age-related cataracts. We investigated whether changes in lens transparency in EphA2 or ephrin-A5 knockout (–/–) mice is related to changes in GJ coupling and lens fluid and ion homeostasis. Immunostaining revealed changes in connexin 50 (Cx50) subcellular localization in EphA2–/– peripheral lens fibers and alteration in aquaporin 0 (Aqp0) staining patterns in ephrin-A5–/– and EphA2–/– inner mature fiber cells. Surprisingly, there was no obvious change in GJ coupling in knockout lenses. However, there were changes in fiber cell membrane conductance and intracellular voltage in knockout lenses from 3-month-old mice. These knockout lenses displayed decreased conductance of mature fiber membranes and were hyperpolarized compared to control lenses. This is the first demonstration that the membrane conductance of lens fibers can be regulated. Together these data suggest that EphA2 may be needed for normal Cx50 localization to the cell membrane and that conductance of lens fiber cells requires normal Eph-ephrin signaling and water channel localization.

Cataract-causing G91del mutant destabilised βA3 heteromers formation linking with structural stability and cellular viability

Background/aimsCongenital cataracts, which are genetically heterogeneous eye disorders, result in visual loss in childhood around the world. CRYBA1/BA3 serves as an abundant structural protein in the lens, and forms homomers and heteromers to maintain lens transparency. In previous study, we identified a common cataract-causing mutation, βA3-glycine at codon 91 (G91del) (c.271–273delGAG), which deleted a highly conserved G91del and led to perinuclear zonular cataract. In this study, we aimed to explore the underlying pathogenic mechanism of G91del mutation.MethodsProtein purification, size-exclusion chromatography, spectroscopy and molecular dynamics simulation assays were used to investigate the effects on the heteromers formation and the protein structural properties of βA3-crystallin caused by G91del mutation. Intracellular βA3-G91del overexpression, MTT (3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide) and cell apoptosis were used to investigate the cellular functions of βA3-G91del.ResultsβA3-crystallin and βB2-crystallin could form heteromers, which have much more stable structures than βA3 homomers. Interestingly, βA3/βB2 heteromers improved their resistance against the thermal stress and the guanidine hydrochloride treatment. However, the pathogenic mutation βA3-G91del destroyed the interaction with βB2, and thereby decreased its structural stability as well as the resistance of thermal or chemical stress. What’s more, the βA3-G91del mutation induced cell apoptosis and escaped from the protection of βB2-crystallin.ConclusionsβA3/βB2 heteromers play an indispensable role in maintaining lens transparency, while the βA3-G91del mutation destabilises heteromers formation with βB2-crystallin, impairs cellular viability and induces cellular apoptosis. These all might contribute to cataract development.

Fundamentals of Diabetic Cataractogenesis and Promising Ways of its Pharmacological Correction

Cataracts in diabetes mellitus lead to decreased visual function and blindness. Cataract surgery for diabetes mellitus has limitations and complications. The search for effective means of conservative cataract therapy continues. The review presents the analysis of data from scientific sources, mainly for 2015–2020 using Internet resources (PubMed, Web of Science, Medline, eLibrary.Ru, Cyberleninka). In the work, diabetic cataractogenesis is presented as a sum of interrelated pathobiochemical processes. The main ones are the polyol pathway of glucose conversion, non-enzymatic glycation and oxidative modification of lens proteins, which are enhanced in diabetes mellitus. The lens has a high protein content. The formation of high molecular weight protein aggregates is of particular importance for the appearance of light scattering zones and a decrease in lens transparency. This review presents data on anti-cataract compounds that affect post-translational crystallin modification, prevent osmotic and oxidative stress in the lens, and exhibit antiglycation properties. This information shows that the search for means of pharmacological correction of cataractogenesis should be carried out among compounds with antioxidant and antiglycation activity.

Mechanosensitive collaboration between integrins and connexins allows nutrient and antioxidant transport into the lens

The delivery of glucose and antioxidants is vital to maintain homeostasis and lens transparency. Here, we report a new mechanism whereby mechanically activated connexin (Cx) hemichannels serve as a transport portal for delivering glucose and glutathione (GSH). Integrin α6β1 in outer cortical lens fiber activated by fluid flow shear stress (FFSS) induced opening of hemichannels. Inhibition of α6 activation prevented hemichannel opening as well as glucose and GSH uptake. The activation of integrin β1, a heterodimeric partner of α6 in the absence of FFSS, increased Cx50 hemichannel opening. Hemichannel activation by FFSS depended on the interaction of integrin α6 and Cx50 C-terminal domain. Moreover, hemichannels in nuclear fiber were unresponsive owing to Cx50 truncation. Taken together, these results show that mechanically activated α6β1 integrin in outer cortical lens fibers leads to opening of hemichannels, which transport glucose and GSH into cortical lens fibers. This study unveils a new transport mechanism that maintains metabolic and antioxidative function of the lens.

“CORRELATION OF GLUCOSE AND CALCIUM IN AQUEOUS HUMOR BETWEEN DIABETIC AND NON-DIABETIC CATARACT PATIENTS''

Cataract is defined as the loss of lens transparency because of opacification of the lens. Based on the causes, cataracts can be classified into age-related cataract, paediatric cataract and cataract due to other causes1. Age-related cataract is the most prevalent type in adults, with the onset between age 45 to 50 years, while in children hereditary and metabolic causes are most common2. Cataract occurs more frequently in low to medium socioeconomic background individuals, and therefore more common in developing countries.3