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.

Immunohistochemical Expression of Alpha (Α) A Crystallin in Senile Degenerative and Non-Cataract Lenses

Aim: Comparative immunohistochemical study of expression of α A Crystallin in non-cataract lenses and age-related cataract lenses in humans. Methodology: This was an observational cross sectional study. There are two groups in this study. Group A comprised of 121 senile degenerative cataract lenses from diagnosed patients. Group B included of 10 non-cataract lenses from patients who underwent surgeries for enucleation due to trauma and retinoblastoma. Lenses were fixed in 10% Buffered Neutral Formalin and processed to make paraffin blocks. Immunohistochemistry (IHC) staining was performed on sections using primary antibody for α A crystallin. Data was analyzed through SPSS software version 24. Results: Immunohistochemical staining of group A showed 80.2% Strong Positive expression while 19.8% showed Intermediate Positive expression of α A Crystallin. 100% Strong Positive expression of α A Crystallin was seen in group B. Comparison of expression of α A Crystallin in two groups showed significant decrease (p<0.001) in expression. Conclusion: Decreased expression of α A Crystallin in IHC stained senile cataract lens indicates the role of structural alterations of lens fibers in pathogenesis of senile cataract. If mechanism involved in causing these alterations can be identified and targeted so that progression of senile cataract may be delayed. Keywords: Immunohistochemistry, α A crystallin expression, senile cataract, Human eye Lens, Lens Fiber.

Some Aspects of Development and Histological Structure of the Visual System of Nothobranchius Guentheri

In this, work some aspects of the development of the visual system of Nothobranchius guentheri at the main stages of ontogenesis were described for the first time. It was possible to establish that the formation of the visual system occurs similarly to other representatives of the order Cyprinodontiformes, but significantly differs in terms of the individual stages of embryogenesis due to the presence of diapause. In the postembryonic period, there is a further increase in the size of the fish’s eyes and head, to the proportions characteristic of adult fish. The histological structure of the eye in adult N. guentheri practically does not differ from most teleost fish living in the same environmental conditions. The study of the structure of the retina showed the heterogeneity of the thickness of the temporal and nasal areas, which indicates the predominant role of peripheral vision. Morphoanatomical measurements of the body and eyes of N. guentheri showed that their correlation was conservative. This indicates an important role of the visual system for the survival of fish in natural conditions, both for the young and adults. In individuals of the older age group, a decrease in the amount of sodium (Na) and an increase in magnesium (Mg) and calcium (Ca) were found in the eye lens. Such changes in the elemental composition of the lens can be a sign of the initial stage of cataractogenesis and disturbances in the metabolism of lens fibers as a result of aging. This allows us to propose N. guentheri as a model for studying the structure, formation, and aging of the visual and nervous systems.

Spontaneous Background and Procedure-Related Microscopic Findings and Common Artifacts in Ocular Tissues of Laboratory Animals in Ocular Studies

Identification of test article–related microscopic findings in ocular toxicology studies requires a working knowledge of the artifacts and procedure-related or background findings commonly encountered in such studies. The objective of this article is to provide a mini-atlas of the artifacts and procedure-related or spontaneous background findings commonly observed in ocular tissues from animals in toxicology studies of ocular drug candidates. Artifacts in the eye are often related to collection or fixation procedures and include swelling and vacuolation of lens fibers, separation of the neuroretina from the retinal pigment epithelium (RPE), and vacuolation of the optic nerve. Common in-life procedure-related findings include intravitreal injection needle tracks in the sclera and ciliary body pars plana and foci of RPE hypertrophy and/or hyperpigmentation at subretinal injection sites. Common background findings include corneal mineralization, uveal mononuclear cell infiltrates, and peripheral displacement of photoreceptor nuclei in the retina. A few uncommon spontaneous background findings that may be confused with test article–related findings, such as bilateral optic atrophy in macaques, are also included.

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.

Ankyrin-B directs membrane tethering of periaxin and is required for maintenance of lens fiber cell hexagonal shape and mechanics

Periaxin (Prx), a PDZ domain protein expressed preferentially in myelinating Schwann cells and lens fibers, plays a key role in membrane scaffolding and cytoarchitecture. Little is known, however, about how Prx is anchored to the plasma membrane. Here we report that ankyrin-B (AnkB), a well-characterized adaptor protein involved in linking the spectrin-actin cytoskeleton to integral membrane proteins, is required for membrane association of Prx in lens fibers and colocalizes with Prx in hexagonal fiber cells. Under AnkB haploinsufficiency, Prx accumulates in the soluble fraction with a concomitant loss from the membrane-enriched fraction of mouse lenses. Moreover, AnkB haploinsufficiency induced age-dependent disruptions in fiber cell hexagonal geometry and radial alignment and decreased compressive stiffness in mouse lenses parallel to the changes observed in Prx null mouse lens. Both AnkB- and Prx-deficient mice exhibit disruptions in membrane organization of the spectrin-actin network and the dystrophin-glycoprotein complex in lens fiber cells. Taken together, these observations reveal that AnkB is required for Prx membrane anchoring and for maintenance of lens fiber cell hexagonal geometry, membrane skeleton organization, and biomechanics.