Mitomycin C-Induced Cell Death in Mouse Lens Epithelial Cells

The effects of melatonin on ionic conductances in a cultured mouse lens epithelial cell line (α-TN4) and in cultured human trabecular meshwork (HTM) cells were measured using the amphotericin perforated patch whole cell voltage-clamp technique. Melatonin stimulated a voltage-dependent Na+-selective current in lens epithelial cells and trabecular meshwork cells. The effects of melatonin were observed at 50 pM and were maximal at 100 μM. Melatonin enhanced activation and inactivation kinetics, but no change was observed in the voltage dependence of activation. The results are consistent with an increase in the total number of ion channels available for activation by membrane depolarization. Melatonin was also found to stimulate a K+-selective current at high doses (1 mM). Melatonin did not affect the inwardly rectifying K+ current or the delayed rectifier type K+ current that has been described in cultured mouse lens epithelial cells. The results show that melatonin specifically stimulated the TTX-insensitive voltage-dependent Na+ current by an apparently novel mechanism.

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Cell kinetic status of mouse lens epithelial cells lacking αA- and αB-crystallin

Molecular and Cellular Biochemistry ◽

10.1023/b:mcbi.0000044365.48900.82 ◽

2004 ◽

Vol 265 (1/2) ◽

pp. 115-122 ◽

Cited By ~ 15

Author(s):

Fang Bai ◽

Jinghua Xi ◽

Ryuji Higashikubo ◽

Usha P. Andley

Keyword(s):

Epithelial Cells ◽

Lens Epithelial Cells ◽

Cell Kinetic ◽

Αb Crystallin ◽

Mouse Lens

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Lens Epithelial Cells Express CD95 and CD95 Ligand Treatment Induces Cell death and DNA Fragmentation in Vitro

European Journal of Ophthalmology ◽

10.1177/112067210301300301 ◽

2003 ◽

Vol 13 (3) ◽

pp. 241-245 ◽

Cited By ~ 5

Author(s):

A. Hueber ◽

C.D. Eichholtz ◽

N. Kociok ◽

J.M. Esser ◽

P.J. Esser

Keyword(s):

Cell Death ◽

Epithelial Cells ◽

Dna Fragmentation ◽

Lens Epithelial Cells ◽

Cd95 Ligand

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Specific features of apoptosis in human lens epithelial cells induced by mitomycin C in vitro

Graefe s Archive for Clinical and Experimental Ophthalmology ◽

10.1007/s004170100332 ◽

2001 ◽

Vol 239 (8) ◽

pp. 613-618 ◽

Cited By ~ 13

Author(s):

J. Jordan ◽

N. Kociok ◽

S. Grisanti ◽

P. Jacobi ◽

J. Esser ◽

...

Keyword(s):

Epithelial Cells ◽

Mitomycin C ◽

Human Lens ◽

Lens Epithelial Cells ◽

Human Lens Epithelial Cells

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Short-Wavelength Blue Light Contributes to Pyroptosis of Human Lens Epithelial Cells (hLECs) by Caspase-1-GSDMD Signalling Axis Activation

10.21203/rs.3.rs-44771/v1 ◽

2020 ◽

Author(s):

Zhaowei Song ◽

Xiaohui Wang ◽

Huazhang Li ◽

Ying Sun ◽

Kexin Liu ◽

...

Keyword(s):

Flow Cytometry ◽

Cell Death ◽

Epithelial Cells ◽

Blue Light ◽

Short Wavelength ◽

Human Lens ◽

Lens Epithelial Cells ◽

Expression Levels ◽

Human Lens Epithelial Cells ◽

Caspase 1

Abstract Backgroud: To examine the effects of short-wavelength blue light (SWBL) on cultured human lens epithelial cells (hLECs). The nosogenesis of cataracts after SWBL exposure was discussed. Methods: HLE-B3 hLECs were divided into 3 groups randomly: A: normal control group, which consisted of hLECs cultured in the dark; B: the caspase-1 inhibitor group; and C: the SWBL exposure group. After the SWBL (2500 lux) irradiation (for 8, 16, 24, and 32 h), the caspase-1 and gasdermin D (GSDMD) expression levels in HLE-B3 hLECs were examined using ELISA, immunofluorescence, and Western blotting analyses. Double-positive staining of HLE-B3 hLECs for activated and inhibited caspase-1 was used to confirm pyroptosis in hLECs by flow cytometry. Results: SWBL can cause cell death in HLE-B3 hLECs, but a caspase-1 inhibitor suppressed cell death. The flow cytometry results also confirmed the does-dependent of short-wavelength blue light irradiation on pyroptotic death of hLECs. Caspase-1 and GSDMD expression levels of all hLECs groups changed with short-wavelength blue light exposure times (8, 16, 24, and 32 h) and were higher in groups B and C than group A. The immunofluorescence results demonstrated that the expression of GSDMD-N was higher in the cell membrane in both the B and C groups than in the A group.Conclusion: The data indicate that SWBL induces pyroptotic programmed cell death by activation of the GSDMD signalling axis in HLE-B3 hLECs. These results provide new insights into the exploitation of new candidates for the prevention of cataracts.

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Changes in lens connexin expression lead to increased gap junctional voltage dependence and conductance

AJP Cell Physiology ◽

10.1152/ajpcell.1995.269.3.c590 ◽

1995 ◽

Vol 269 (3) ◽

pp. C590-C600 ◽

Cited By ~ 32

Author(s):

P. J. Donaldson ◽

Y. Dong ◽

M. Roos ◽

C. Green ◽

D. A. Goodenough ◽

...

Keyword(s):

Epithelial Cells ◽

Gap Junction ◽

Voltage Dependence ◽

Expression Patterns ◽

Fiber Cell ◽

Lens Epithelial Cells ◽

Connexin Expression ◽

Fiber Cells ◽

Gap Junctional ◽

Mouse Lens

The differentiation of mouse lens epithelial cells into fiber cells is a useful model for studying the changes of the electrical properties of gap junction (cell-to-cell) channels that are induced by an alteration in connexin expression patterns. In this model, cuboidal lens epithelial cells differentiate into elongated fiber cells, and the expression of connexin43 (Cx43) in the epithelial cells is replaced with the production of high levels of Cx50 and Cx46 in the fiber cells. We now report a new procedure to isolate mouse lens fiber cell pairs suitable for double whole cell patch-clamp analysis. Analysis was also performed for fiberlike cell pairs differentiated from epithelial cells in culture. Voltage dependence and unitary conductance of fiber cell gap junction channels were determined and compared with the corresponding values previously measured for the channels joining lens epithelial cells and for lens connexin channels formed in Xenopus oocyte pairs. Our results support a differentiation-induced shift toward stronger gap junctional voltage dependence and larger unitary conductances in the fiber cells. Our data further reflect a balanced functional contribution of Cx50 and Cx46 in the fiber cell-to-cell channels rather than a predominance of a single connexin.

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Factors modulating mouse lens epithelial cell morphology with differentiation and development of a lentoid structure in vitro

Development ◽

10.1242/dev.99.1.25 ◽

1987 ◽

Vol 99 (1) ◽

pp. 25-32

Author(s):

A.L. Muggleton-Harris ◽

N. Higbee

Keyword(s):

Epithelial Cells ◽

Collagen Gel ◽

Cell Aggregation ◽

Lens Epithelial Cell ◽

Lens Capsule ◽

Lens Epithelial Cells ◽

Collagen Membrane ◽

Mouse Lens

The morphological and cellular changes that occur with differentiation and development of a lentoid structure from cultured mouse lens epithelial cells have been found to be dependent on the presence of lens capsule in association with the cells. The development of the ‘lentoid body’ is a multiphase process involving cell replication, synthesis of mucosubstances and a basement collagen membrane, cell aggregation and differentiation. Stage-specific synthesis of lens proteins confirms that the genes regulating normal differentiation in vivo are operating in the in vitro system. The hydrated collagen gel studies described in this report demonstrate that the cuboidal morphology and apical-basal polarity of the lens epithelial cells are dependent on their relationship with the lens capsule. Following a replicative phase the cells assume a mesenchyme-like morphology and migrate into the gel. Trypsinized cells freed from the lens capsule replicate but form colonies on the surface of the gel. The implications of these results are discussed with respect to previous observations made on normal lens development and the abnormalities associated with the congenital cataractous embryonic lens.

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