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Glutamate Stimulation Dysregulates AMPA Receptors-Induced Signal Transduction Pathway in Leber’s Inherited Optic Neuropathy Patient-Specific hiPSC-Derived Retinal Ganglion Cells

The mitochondrial genetic disorder, Leber’s hereditary optic neuropathy (LHON), is caused by a mutation in MT-ND4 gene, encoding NADH dehydrogenase subunit 4. It leads to the progressive death of retinal ganglion cells (RGCs) and causes visual impairment or even blindness. However, the precise mecha...

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Autores principales: Yang, Yi-Ping, Nguyen, Phan Nguyen Nhi, Lin, Tai-Chi, Yarmishyn, Aliaksandr A., Chen, Wun-Syuan, Hwang, De-Kuang, Chiou, Guang-Yuh, Lin, Tzu-Wei, Chien, Chian-Shiu, Tsai, Ching-Yao, Chiou, Shih-Hwa, Chen, Shih-Jen, Peng, Chi-Hsien, Hsu, Chih-Chien
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6627514/
https://www.ncbi.nlm.nih.gov/pubmed/31234430
http://dx.doi.org/10.3390/cells8060625
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author Yang, Yi-Ping
Nguyen, Phan Nguyen Nhi
Lin, Tai-Chi
Yarmishyn, Aliaksandr A.
Chen, Wun-Syuan
Hwang, De-Kuang
Chiou, Guang-Yuh
Lin, Tzu-Wei
Chien, Chian-Shiu
Tsai, Ching-Yao
Chiou, Shih-Hwa
Chen, Shih-Jen
Peng, Chi-Hsien
Hsu, Chih-Chien
author_facet Yang, Yi-Ping
Nguyen, Phan Nguyen Nhi
Lin, Tai-Chi
Yarmishyn, Aliaksandr A.
Chen, Wun-Syuan
Hwang, De-Kuang
Chiou, Guang-Yuh
Lin, Tzu-Wei
Chien, Chian-Shiu
Tsai, Ching-Yao
Chiou, Shih-Hwa
Chen, Shih-Jen
Peng, Chi-Hsien
Hsu, Chih-Chien
author_sort Yang, Yi-Ping
collection PubMed
description The mitochondrial genetic disorder, Leber’s hereditary optic neuropathy (LHON), is caused by a mutation in MT-ND4 gene, encoding NADH dehydrogenase subunit 4. It leads to the progressive death of retinal ganglion cells (RGCs) and causes visual impairment or even blindness. However, the precise mechanisms of LHON disease penetrance and progression are not completely elucidated. Human-induced pluripotent stem cells (hiPSCs) offer unique opportunities to investigate disease-relevant phenotypes and regulatory mechanisms underlying LHON pathogenesis at the cellular level. In this study, we successfully generated RGCs by differentiation of LHON patient-specific hiPSCs. We modified the protocol of differentiation to obtain a more enriched population of single-cell RGCs for LHON study. Based on assessing morphology, expression of specific markers and electrophysiological activity, we found that LHON-specific hiPSC-derived were more defective in comparison with normal wild-type RGCs. Based on our previous study, whereby by using microarray analysis we identified that the components of glutamatergic synapse signaling pathway were significantly downregulated in LHON-specific RGCs, we focused our study on glutamate-associated α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) receptors. We found that the protein expression levels of the subunits of the AMPA receptor, GluR1 and GluR2, and their associated scaffold proteins were decreased in LHON-RGCs. By performing the co-immunoprecipitation assay, we found several differences in the efficiencies of interaction between AMPA subunits and scaffold proteins between normal and LHON-specific RGCs.
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spelling pubmed-66275142019-07-23 Glutamate Stimulation Dysregulates AMPA Receptors-Induced Signal Transduction Pathway in Leber’s Inherited Optic Neuropathy Patient-Specific hiPSC-Derived Retinal Ganglion Cells Yang, Yi-Ping Nguyen, Phan Nguyen Nhi Lin, Tai-Chi Yarmishyn, Aliaksandr A. Chen, Wun-Syuan Hwang, De-Kuang Chiou, Guang-Yuh Lin, Tzu-Wei Chien, Chian-Shiu Tsai, Ching-Yao Chiou, Shih-Hwa Chen, Shih-Jen Peng, Chi-Hsien Hsu, Chih-Chien Cells Article The mitochondrial genetic disorder, Leber’s hereditary optic neuropathy (LHON), is caused by a mutation in MT-ND4 gene, encoding NADH dehydrogenase subunit 4. It leads to the progressive death of retinal ganglion cells (RGCs) and causes visual impairment or even blindness. However, the precise mechanisms of LHON disease penetrance and progression are not completely elucidated. Human-induced pluripotent stem cells (hiPSCs) offer unique opportunities to investigate disease-relevant phenotypes and regulatory mechanisms underlying LHON pathogenesis at the cellular level. In this study, we successfully generated RGCs by differentiation of LHON patient-specific hiPSCs. We modified the protocol of differentiation to obtain a more enriched population of single-cell RGCs for LHON study. Based on assessing morphology, expression of specific markers and electrophysiological activity, we found that LHON-specific hiPSC-derived were more defective in comparison with normal wild-type RGCs. Based on our previous study, whereby by using microarray analysis we identified that the components of glutamatergic synapse signaling pathway were significantly downregulated in LHON-specific RGCs, we focused our study on glutamate-associated α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) receptors. We found that the protein expression levels of the subunits of the AMPA receptor, GluR1 and GluR2, and their associated scaffold proteins were decreased in LHON-RGCs. By performing the co-immunoprecipitation assay, we found several differences in the efficiencies of interaction between AMPA subunits and scaffold proteins between normal and LHON-specific RGCs. MDPI 2019-06-21 /pmc/articles/PMC6627514/ /pubmed/31234430 http://dx.doi.org/10.3390/cells8060625 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Yang, Yi-Ping
Nguyen, Phan Nguyen Nhi
Lin, Tai-Chi
Yarmishyn, Aliaksandr A.
Chen, Wun-Syuan
Hwang, De-Kuang
Chiou, Guang-Yuh
Lin, Tzu-Wei
Chien, Chian-Shiu
Tsai, Ching-Yao
Chiou, Shih-Hwa
Chen, Shih-Jen
Peng, Chi-Hsien
Hsu, Chih-Chien
Glutamate Stimulation Dysregulates AMPA Receptors-Induced Signal Transduction Pathway in Leber’s Inherited Optic Neuropathy Patient-Specific hiPSC-Derived Retinal Ganglion Cells
title Glutamate Stimulation Dysregulates AMPA Receptors-Induced Signal Transduction Pathway in Leber’s Inherited Optic Neuropathy Patient-Specific hiPSC-Derived Retinal Ganglion Cells
title_full Glutamate Stimulation Dysregulates AMPA Receptors-Induced Signal Transduction Pathway in Leber’s Inherited Optic Neuropathy Patient-Specific hiPSC-Derived Retinal Ganglion Cells
title_fullStr Glutamate Stimulation Dysregulates AMPA Receptors-Induced Signal Transduction Pathway in Leber’s Inherited Optic Neuropathy Patient-Specific hiPSC-Derived Retinal Ganglion Cells
title_full_unstemmed Glutamate Stimulation Dysregulates AMPA Receptors-Induced Signal Transduction Pathway in Leber’s Inherited Optic Neuropathy Patient-Specific hiPSC-Derived Retinal Ganglion Cells
title_short Glutamate Stimulation Dysregulates AMPA Receptors-Induced Signal Transduction Pathway in Leber’s Inherited Optic Neuropathy Patient-Specific hiPSC-Derived Retinal Ganglion Cells
title_sort glutamate stimulation dysregulates ampa receptors-induced signal transduction pathway in leber’s inherited optic neuropathy patient-specific hipsc-derived retinal ganglion cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6627514/
https://www.ncbi.nlm.nih.gov/pubmed/31234430
http://dx.doi.org/10.3390/cells8060625
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