Verification and spatial mapping of TRPV1 and TRPV4 expression in the embryonic and adult mouse lens

Nakazawa, Yosuke; Donaldson, Paul J.; Petrova, Rosica S.

doi:10.1016/j.exer.2019.107707

Cited by 32 publications

(31 citation statements)

References 37 publications

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“…S2). As in other epithelia, 42 the subcellular distribution of TRPV4-ir included plasma membrane and cytosolic signals. Trpv4 −/− corneas stained for TRPV4 showed little labeling and few discernible changes in overall architecture ( Fig.…”

Section: Trpv4 the Dominant Thermotrp Isoform In Cecs Is Distributementioning

confidence: 77%

Polymodal Sensory Transduction in Mouse Corneal Epithelial Cells

Lapajne

Lakk

Yarishkin

et al. 2020

Invest. Ophthalmol. Vis. Sci.

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PURPOSE. Contact lenses, osmotic stressors, and chemical burns may trigger severe discomfort and vision loss by damaging the cornea, but the signaling mechanisms used by corneal epithelial cells (CECs) to sense extrinsic stressors are not well understood. We therefore investigated the mechanisms of swelling, temperature, strain, and chemical transduction in mouse CECs. METHODS. Intracellular calcium imaging in conjunction with electrophysiology, pharmacology, transcript analysis, immunohistochemistry, and bioluminescence assays of adenosine triphosphate (ATP) release were used to track mechanotransduction in dissociated CECs and epithelial sheets isolated from the mouse cornea. RESULTS. The transient receptor potential vanilloid (TRPV) transcriptome in the mouse corneal epithelium is dominated by Trpv4, followed by Trpv2, Trpv3, and low levels of Trpv1 mRNAs. TRPV4 protein was localized to basal and intermediate epithelial strata, keratocytes, and the endothelium in contrast to the cognate TRPV1, which was confined to intraepithelial afferents and a sparse subset of CECs. The TRPV4 agonist GSK1016790A induced cation influx and calcium elevations, which were abolished by the selective blocker HC067047. Hypotonic solutions, membrane strain, and moderate heat elevated [Ca 2+ ] CEC with swelling-and temperature-, but not strain-evoked signals, sensitive to HC067047. GSK1016790A and swelling evoked calcium-dependent ATP release, which was suppressed by HC067027 and the hemichannel blocker probenecid. CONCLUSIONS. These results demonstrate that cation influx via TRPV4 transduces osmotic and thermal but not strain inputs to CECs and promotes hemichannel-dependent ATP release. The TRPV4-hemichannel-ATP signaling axis might modulate corneal pain induced by excessive mechanical, osmotic, and chemical stimulation.

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Section: Trpv4 the Dominant Thermotrp Isoform In Cecs Is Distributementioning

confidence: 77%

Polymodal Sensory Transduction in Mouse Corneal Epithelial Cells

Lapajne

Lakk

Yarishkin

et al. 2020

Invest. Ophthalmol. Vis. Sci.

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“… 41 – 43 In the lens, TRPV4 and TRPV1 channels have been shown to not only reciprocally regulate the hydrostatic pressure generated by water flow through gap junction channels 6 but also transduce changes to the magnitude of this pressure gradient induced by pharmacologically modulating the zonular tension applied to the lens. 7 Furthermore, changes in zonular tension induced either mechanically by cutting the zonules 44 or pharmacologically via application of tropicamide or pilocarpine also altered the subcellular location of TRPV1 and TRPV4 (data presented at the Sixth International Conference on the Lens by Nakazawa et al, unpublished data). Taken together, these observations suggest that in addition to the modulation of the osmotic gradients that drive the transport of water, 6 , 45 changes to P H2O are also required to maintain the gradient in hydrostatic pressure that has been measured in all lenses studied to date.…”

Section: Discussionmentioning

confidence: 97%

Changes to Zonular Tension Alters the Subcellular Distribution of AQP5 in Regions of Influx and Efflux of Water in the Rat Lens

Petrova

Bavana

Zhao

et al. 2020

Invest. Ophthalmol. Vis. Sci.

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The lens uses circulating fluxes of ions and water that enter the lens at both poles and exit at the equator to maintain its optical properties. We have mapped the subcellular distribution of the lens aquaporins (AQP0, AQP1, and AQP5) in these water influx and efflux zones and investigated how their membrane location is affected by changes in tension applied to the lens by the zonules. METHODS. Immunohistochemistry using AQP antibodies was performed on axial sections obtained from rat lenses that had been removed from the eye and then fixed or were fixed in situ to maintain zonular tension. Zonular tension was pharmacologically modulated by applying either tropicamide (increased) or pilocarpine (decreased). AQP labeling was visualized using confocal microscopy. RESULTS. Modulation of zonular tension had no effect on AQP1 or AQP0 labeling in either the water efflux or influx zones. In contrast, AQP5 labeling changed from membranous to cytoplasmic in response to both mechanical and pharmacologically induced reductions in zonular tension in both the efflux zone and anterior (but not posterior) influx zone associated with the lens sutures. CONCLUSIONS. Altering zonular tension dynamically regulates the membrane trafficking of AQP5 in the efflux and anterior influx zones to potentially change the magnitude of circulating water fluxes in the lens.

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“…Several factors may be involved in age-related nuclear cataracts: connexins and aquaporins (AQPs) related to metabolite transport in the lens, transient receptor potential cation channel subfamily V member 1 (TRPV1) and transient receptor potential cation channel subfamily V member 4 (TRPV4) proteins involved in lens ion transport [ 65 ], the post-translational modification of various crystallin proteins in the deep lens cortex and nucleus [ 66 , 67 ], and the oxidation of the lens nucleus due to antioxidants’ inability to reach the lens nucleus [ 68 ]. We plan to investigate various post-translational modifications, the effects of culture temperature on the proteins related to intercellular transport, two- and three-dimensional cultures, and lens organ cultures.…”

Section: Discussionmentioning

confidence: 99%

Effect of a Lens Protein in Low-Temperature Culture of Novel Immortalized Human Lens Epithelial Cells (iHLEC-NY2)

Yamamoto

Takeda

Hatsusaka

et al. 2020

Cells

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The prevalence of nuclear cataracts was observed to be significantly higher among residents of tropical and subtropical regions compared to those of temperate and subarctic regions. We hypothesized that elevated environmental temperatures may pose a risk of nuclear cataract development. The results of our in silico simulation revealed that in temperate and tropical regions, the human lens temperature ranges from 35.0 °C to 37.5 °C depending on the environmental temperature. The medium temperature changes during the replacement regularly in the cell culture experiment were carefully monitored using a sensor connected to a thermometer and showed a decrease of 1.9 °C, 3.0 °C, 1.7 °C, and 0.1 °C, after 5 min when setting the temperature of the heat plate device at 35.0 °C, 37.5 °C, 40.0 °C, and 42.5 °C, respectively. In the newly created immortalized human lens epithelial cell line clone NY2 (iHLEC-NY2), the amounts of RNA synthesis of αA crystallin, protein expression, and amyloid β (Aβ)1-40 secreted into the medium were increased at the culture temperature of 37.5 °C compared to 35.0 °C. In short-term culture experiments, the secretion of Aβ1-40 observed in cataracts was increased at 37.5 °C compared to 35.0 °C, suggesting that the long-term exposure to a high-temperature environment may increase the risk of cataracts.

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Verification and spatial mapping of TRPV1 and TRPV4 expression in the embryonic and adult mouse lens

Cited by 32 publications

References 37 publications

Polymodal Sensory Transduction in Mouse Corneal Epithelial Cells

Polymodal Sensory Transduction in Mouse Corneal Epithelial Cells

Changes to Zonular Tension Alters the Subcellular Distribution of AQP5 in Regions of Influx and Efflux of Water in the Rat Lens

Effect of a Lens Protein in Low-Temperature Culture of Novel Immortalized Human Lens Epithelial Cells (iHLEC-NY2)

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