Induction of Neuronal Morphology in the 661W Cone Photoreceptor Cell Line with Staurosporine

Thompson, Alex F.; Crowe, Megan E.; Lieven, Christopher J.; Levin, Leonard A.

doi:10.1371/journal.pone.0145270

Cited by 17 publications

(17 citation statements)

References 30 publications

(42 reference statements)

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“…Upon adherence to the surface, 661W cells flatten out and start contacting the neighboring cells through their neuronal processes, usually attaching to multiple cells at a time. Treatment of 661W and RGC-5 cells with staurosporine for 24 hours resulted in differentiation into cells with prominent neuronal morphology 24 . Differentiated cells showed larger and more number of cytoplasmic projections/neurites, which could be visualized more clearly upon staining with acetylated tubulin-specific antibody (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…However, morphologically, these cells show processes generally seen in neuronal cells and do not form outer segment-like membranes seen in photoreceptor cells 22 . Treatment of these cells with staurosporine (STSN) induces differentiation into neuronal cells that show increased number and branching of neurites per cell 24 . Although expression of several molecular markers for cone and rod photoreceptors and retinal pigment epithelium (RPE), has been tested, the markers for RGCs have not been examined in this cell line.…”

Section: Introductionmentioning

confidence: 99%

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661W is a retinal ganglion precursor-like cell line in which glaucoma-associated optineurin mutants induce cell death selectively

Sayyad

Sirohi

Radha

et al. 2017

Sci Rep

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A photoreceptor cell line, 661W, derived from a mouse retinal tumor that expresses several markers of cone photoreceptor cells has been described earlier. However, these cells can be differentiated into neuronal cells. Here, we report that this cell line expressed certain markers specific to retinal ganglion cells such as Rbpms, Brn3b (Pou4f2), Brn3c (Pou4f3), Thy1 and γ-synuclein (Sncg), and some other markers of neuronal cells (beta-III tubulin, NeuN and MAP2). These cells also expressed Opn1mw, a cone-specific marker and nestin, a marker for neural precursor cells. Two glaucoma-associated mutants of OPTN, E50K and M98K, but not an amyotrophic lateral sclerosis-associated mutant, E478G, induced cell death selectively in 661W cells. However, in a motor neuron cell line, NSC34, E478G mutant of OPTN but not E50K and M98K induced cell death. We conclude that 661W is a retinal ganglion precursor-like cell line, which shows properties of both retinal ganglion and photoreceptor cells. We suggest that these cells could be utilized for exploring the mechanisms of cell death induction and cytoprotection relevant for glaucoma pathogenesis. RGC-5 cell line which probably arose from 661W cells showed expression of essentially the same markers of retinal ganglion cells and neuronal cells as seen in 661W cells.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

661W is a retinal ganglion precursor-like cell line in which glaucoma-associated optineurin mutants induce cell death selectively

Sayyad

Sirohi

Radha

et al. 2017

Sci Rep

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“…RGC-5 cells (gift of Neeraj Agarwal, Ph.D.) are a cell line derived from retina neurons that despite their name are most similar to the 661W cone photoreceptor cell line, a type of retinal neuron (Al-Ubaidi, 2014; Krishnamoorthy et al, 2013; Thompson et al, 2015). They were incubated in 20 μM DSSA-1 for 1 hour.…”

Section: Methodsmentioning

confidence: 99%

Intracellular disulfide reduction by phosphine-borane complexes: Mechanism of action for neuroprotection

Niemuth

Thompson

Crowe

et al. 2016

Neurochemistry International

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Phosphine-borane complexes are novel cell-permeable drugs that protect neurons from axonal injury in vitro and in vivo. These drugs activate the extracellular signal-regulated kinases 1/2 (ERK1/2) cell survival pathway and are therefore neuroprotective, but do not scavenge superoxide. In order to understand the interaction between superoxide signaling of neuronal death and the action of phosphine-borane complexes, their biochemical activity in cell-free and in vitro assays was studied by electron paramagnetic resonance (EPR) spectrometry and using an intracellular dithiol reporter that becomes fluorescent when its disulfide bond is cleaved. These studies demonstrated that bis(3-propionic acid methyl ester) phenylphosphine-borane complex (PB1) and (3-propionic acid methyl ester) diphenylphosphine-borane complex (PB2) are potent intracellular disulfide reducing agents which are cell permeable. EPR and pharmacological studies demonstrated reducing activity but not scavenging of superoxide. Given that phosphine-borane complexes reduce cell injury from mitochondrial superoxide generation but do not scavenge superoxide, this implies a mechanism where an intracellular superoxide burst induces downstream formation of protein disulfides. The redox-dependent cleavage of the disulfides is therefore a novel mechanism of neuroprotection.

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“…The specificity of this commercial Brn3 antibody was not tested by immunoblotting ( 84 ). Interestingly, 661W as well as RGC-5 cells can be differentiated into cells with prominent neuronal morphology ( 85 ). Recently, both 661W and RGC-5 cell lines have been re-characterized ( 86 ).…”

Section: Model Systems To Study Glaucoma Pathogenesismentioning

confidence: 99%

Altered Functions and Interactions of Glaucoma-Associated Mutants of Optineurin

Swarup

Sayyad

2018

Front. Immunol.

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Optineurin (OPTN) is an adaptor protein that is involved in mediating a variety of cellular processes such as signaling, vesicle trafficking, and autophagy. Certain mutations in OPTN (gene OPTN) are associated with primary open angle glaucoma, a leading cause of irreversible blindness, and amyotrophic lateral sclerosis, a fatal motor neuron disease. Glaucoma-associated mutations of OPTN are mostly missense mutations. OPTN mediates its functions by interacting with various proteins and altered interactions of OPTN mutants with various proteins primarily contribute to functional defects. It interacts with Rab8, myosin VI, Huntigtin, TBC1D17, and transferrin receptor to mediate various membrane vesicle trafficking pathways. It is an autophagy receptor that mediates cargo-selective as well as non-selective autophagy. Glaucoma-associated mutants of OPTN, E50K, and M98K, cause defective vesicle trafficking, autophagy, and signaling that contribute to death of retinal ganglion cells (RGCs). Transgenic mice expressing E50K-OPTN show loss of RGCs and persistent reactive gliosis. TBK1 protein kinase, which mediates E50K-OPTN and M98K-OPTN induced cell death, is emerging as a potential drug target. Autoimmunity has been implicated in glaucoma but involvement of OPTN or its mutants in autoimmnity has not been explored. In this review, we highlight the main functions of OPTN and how glaucoma-associated mutants alter these functions. We also discuss some of the controversies, such as the role of OPTN in signaling to transcription factor NF-κB, interferon signaling, and use of RGC-5 cell line as a cell culture model.

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Induction of Neuronal Morphology in the 661W Cone Photoreceptor Cell Line with Staurosporine

Cited by 17 publications

References 30 publications

661W is a retinal ganglion precursor-like cell line in which glaucoma-associated optineurin mutants induce cell death selectively

661W is a retinal ganglion precursor-like cell line in which glaucoma-associated optineurin mutants induce cell death selectively

Intracellular disulfide reduction by phosphine-borane complexes: Mechanism of action for neuroprotection

Altered Functions and Interactions of Glaucoma-Associated Mutants of Optineurin

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