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The GJA8 allele encoding CX50I247M is a rare polymorphism, not a cataract-causing mutation
PURPOSE: The aim of this study was the genetic, cellular, and physiological characterization of a connexin50 (CX50) variant identified in a child with congenital cataracts. METHODS: Lens material from surgery was collected and used for cDNA production. Genomic DNA was prepared from blood obtained fr...
Autores principales: | , , , , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
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Molecular Vision
2009
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2743802/ https://www.ncbi.nlm.nih.gov/pubmed/19756179 |
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author | Graw, Jochen Schmidt, Werner Minogue, Peter J. Rodriguez, Jessica Tong, Jun-Jie Klopp, Norman Illig, Thomas Ebihara, Lisa Berthoud, Viviana M. Beyer, Eric C. |
author_facet | Graw, Jochen Schmidt, Werner Minogue, Peter J. Rodriguez, Jessica Tong, Jun-Jie Klopp, Norman Illig, Thomas Ebihara, Lisa Berthoud, Viviana M. Beyer, Eric C. |
author_sort | Graw, Jochen |
collection | PubMed |
description | PURPOSE: The aim of this study was the genetic, cellular, and physiological characterization of a connexin50 (CX50) variant identified in a child with congenital cataracts. METHODS: Lens material from surgery was collected and used for cDNA production. Genomic DNA was prepared from blood obtained from the proband and her parents. PCR amplified DNA fragments were sequenced and characterized by restriction digestion. Connexin protein distribution was studied by immunofluorescence in transiently transfected HeLa cells. Formation of functional channels was assessed by two-microelectrode voltage-clamp in cRNA-injected Xenopus oocytes. RESULTS: Ophthalmologic examination showed that the proband suffered from bilateral white, diffuse cataracts, but the parents were free of lens opacities. Direct sequencing of the PCR product produced from lens cDNA showed that the proband was heterozygous for a G>T transition at position 741 of the GJA8 gene, encoding the exchange of methionine for isoleucine at position 247 of CX50 (CX50I247M). The mutation was confirmed in the genomic DNA, but it was also present in the unaffected mother. When expressed in HeLa cells, both wild type CX50 and CX50I247M formed gap junction plaques. Both CX50 and CX50I247M induced gap junctional currents in pairs of Xenopus oocytes. CONCLUSIONS: Although the CX50I247M substitution has previously been suggested to cause cataracts, our genetic, cellular, and electrophysiological data suggest that this allele more likely represents a rare silent, polymorphic variant. |
format | Text |
id | pubmed-2743802 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2009 |
publisher | Molecular Vision |
record_format | MEDLINE/PubMed |
spelling | pubmed-27438022009-09-15 The GJA8 allele encoding CX50I247M is a rare polymorphism, not a cataract-causing mutation Graw, Jochen Schmidt, Werner Minogue, Peter J. Rodriguez, Jessica Tong, Jun-Jie Klopp, Norman Illig, Thomas Ebihara, Lisa Berthoud, Viviana M. Beyer, Eric C. Mol Vis Research Article PURPOSE: The aim of this study was the genetic, cellular, and physiological characterization of a connexin50 (CX50) variant identified in a child with congenital cataracts. METHODS: Lens material from surgery was collected and used for cDNA production. Genomic DNA was prepared from blood obtained from the proband and her parents. PCR amplified DNA fragments were sequenced and characterized by restriction digestion. Connexin protein distribution was studied by immunofluorescence in transiently transfected HeLa cells. Formation of functional channels was assessed by two-microelectrode voltage-clamp in cRNA-injected Xenopus oocytes. RESULTS: Ophthalmologic examination showed that the proband suffered from bilateral white, diffuse cataracts, but the parents were free of lens opacities. Direct sequencing of the PCR product produced from lens cDNA showed that the proband was heterozygous for a G>T transition at position 741 of the GJA8 gene, encoding the exchange of methionine for isoleucine at position 247 of CX50 (CX50I247M). The mutation was confirmed in the genomic DNA, but it was also present in the unaffected mother. When expressed in HeLa cells, both wild type CX50 and CX50I247M formed gap junction plaques. Both CX50 and CX50I247M induced gap junctional currents in pairs of Xenopus oocytes. CONCLUSIONS: Although the CX50I247M substitution has previously been suggested to cause cataracts, our genetic, cellular, and electrophysiological data suggest that this allele more likely represents a rare silent, polymorphic variant. Molecular Vision 2009-09-14 /pmc/articles/PMC2743802/ /pubmed/19756179 Text en http://creativecommons.org/licenses/by/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Article Graw, Jochen Schmidt, Werner Minogue, Peter J. Rodriguez, Jessica Tong, Jun-Jie Klopp, Norman Illig, Thomas Ebihara, Lisa Berthoud, Viviana M. Beyer, Eric C. The GJA8 allele encoding CX50I247M is a rare polymorphism, not a cataract-causing mutation |
title | The GJA8 allele encoding CX50I247M is a rare polymorphism, not a cataract-causing mutation |
title_full | The GJA8 allele encoding CX50I247M is a rare polymorphism, not a cataract-causing mutation |
title_fullStr | The GJA8 allele encoding CX50I247M is a rare polymorphism, not a cataract-causing mutation |
title_full_unstemmed | The GJA8 allele encoding CX50I247M is a rare polymorphism, not a cataract-causing mutation |
title_short | The GJA8 allele encoding CX50I247M is a rare polymorphism, not a cataract-causing mutation |
title_sort | gja8 allele encoding cx50i247m is a rare polymorphism, not a cataract-causing mutation |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2743802/ https://www.ncbi.nlm.nih.gov/pubmed/19756179 |
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