Design of a novel quantitative PCR (QPCR)-based protocol for genotyping mice carrying the neuroprotective Wallerian degeneration slow (<it>Wld</it><sup><it>s</it></sup>) gene
<p>Abstract</p> <p>Background</p> <p>Mice carrying the spontaneous genetic mutation known as Wallerian degeneration slow (<it>Wld</it><sup><it>s</it></sup>) have a unique neuroprotective phenotype, where axonal and synaptic compartmen...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2007-10-01
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| Series: | Molecular Neurodegeneration |
| Online Access: | http://www.molecularneurodegeneration.com/content/2/1/21 |
| _version_ | 1811278626973286400 |
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| author | Coleman Michael P Shipston Michael J Chen Philip E MacDonald Stephen HF Wishart Thomas M Gillingwater Thomas H Ribchester Richard R |
| author_facet | Coleman Michael P Shipston Michael J Chen Philip E MacDonald Stephen HF Wishart Thomas M Gillingwater Thomas H Ribchester Richard R |
| author_sort | Coleman Michael P |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>Mice carrying the spontaneous genetic mutation known as Wallerian degeneration slow (<it>Wld</it><sup><it>s</it></sup>) have a unique neuroprotective phenotype, where axonal and synaptic compartments of neurons are protected from degeneration following a wide variety of physical, toxic and inherited disease-inducing stimuli. This remarkable phenotype has been shown to delay onset and progression in several mouse models of neurodegenerative disease, suggesting that <it>Wld</it><sup><it>s</it></sup>-mediated neuroprotection may assist in the identification of novel therapeutic targets. As a result, cross-breeding of <it>Wld</it><sup><it>s </it></sup>mice with mouse models of neurodegenerative diseases is used increasingly to understand the roles of axon and synapse degeneration in disease. However, the phenotype shows strong gene-dose dependence so it is important to distinguish offspring that are homozygous or heterozygous for the mutation. Since the <it>Wld</it><sup><it>s </it></sup>mutation comprises a triplication of a region already present in the mouse genome, the most stringent way to quantify the number of mutant <it>Wld</it><sup><it>s </it></sup>alleles is using copy number. Current approaches to genotype <it>Wld</it><sup><it>s </it></sup>mice are based on either Southern blots or pulsed field gel electrophoresis, neither of which are as rapid or efficient as quantitative PCR (QPCR).</p> <p>Results</p> <p>We have developed a rapid, robust and efficient genotyping method for <it>Wld</it><sup><it>s </it></sup>using QPCR. This approach differentiates, based on copy number, homozygous and heterozygous <it>Wld</it><sup><it>s </it></sup>mice from wild-type mice and each other. We show that this approach can be used to genotype mice carrying the spontaneous <it>Wld</it><sup><it>s </it></sup>mutation as well as animals expressing the <it>Wld</it><sup><it>s </it></sup>transgene.</p> <p>Conclusion</p> <p>We have developed a QPCR genotyping method that permits rapid and effective genotyping of <it>Wld</it><sup><it>s </it></sup>copy number. This technique will be of particular benefit in studies where <it>Wld</it><sup><it>s </it></sup>mice are cross-bred with other mouse models of neurodegenerative disease in order to understand the neuroprotective processes conferred by the <it>Wld</it><sup><it>s </it></sup>mutation.</p> |
| first_indexed | 2024-04-13T00:39:14Z |
| format | Article |
| id | doaj.art-af35a258da52453db7019f55d1c39452 |
| institution | Directory Open Access Journal |
| issn | 1750-1326 |
| language | English |
| last_indexed | 2024-04-13T00:39:14Z |
| publishDate | 2007-10-01 |
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| series | Molecular Neurodegeneration |
| spelling | doaj.art-af35a258da52453db7019f55d1c394522022-12-22T03:10:14ZengBMCMolecular Neurodegeneration1750-13262007-10-01212110.1186/1750-1326-2-21Design of a novel quantitative PCR (QPCR)-based protocol for genotyping mice carrying the neuroprotective Wallerian degeneration slow (<it>Wld</it><sup><it>s</it></sup>) geneColeman Michael PShipston Michael JChen Philip EMacDonald Stephen HFWishart Thomas MGillingwater Thomas HRibchester Richard R<p>Abstract</p> <p>Background</p> <p>Mice carrying the spontaneous genetic mutation known as Wallerian degeneration slow (<it>Wld</it><sup><it>s</it></sup>) have a unique neuroprotective phenotype, where axonal and synaptic compartments of neurons are protected from degeneration following a wide variety of physical, toxic and inherited disease-inducing stimuli. This remarkable phenotype has been shown to delay onset and progression in several mouse models of neurodegenerative disease, suggesting that <it>Wld</it><sup><it>s</it></sup>-mediated neuroprotection may assist in the identification of novel therapeutic targets. As a result, cross-breeding of <it>Wld</it><sup><it>s </it></sup>mice with mouse models of neurodegenerative diseases is used increasingly to understand the roles of axon and synapse degeneration in disease. However, the phenotype shows strong gene-dose dependence so it is important to distinguish offspring that are homozygous or heterozygous for the mutation. Since the <it>Wld</it><sup><it>s </it></sup>mutation comprises a triplication of a region already present in the mouse genome, the most stringent way to quantify the number of mutant <it>Wld</it><sup><it>s </it></sup>alleles is using copy number. Current approaches to genotype <it>Wld</it><sup><it>s </it></sup>mice are based on either Southern blots or pulsed field gel electrophoresis, neither of which are as rapid or efficient as quantitative PCR (QPCR).</p> <p>Results</p> <p>We have developed a rapid, robust and efficient genotyping method for <it>Wld</it><sup><it>s </it></sup>using QPCR. This approach differentiates, based on copy number, homozygous and heterozygous <it>Wld</it><sup><it>s </it></sup>mice from wild-type mice and each other. We show that this approach can be used to genotype mice carrying the spontaneous <it>Wld</it><sup><it>s </it></sup>mutation as well as animals expressing the <it>Wld</it><sup><it>s </it></sup>transgene.</p> <p>Conclusion</p> <p>We have developed a QPCR genotyping method that permits rapid and effective genotyping of <it>Wld</it><sup><it>s </it></sup>copy number. This technique will be of particular benefit in studies where <it>Wld</it><sup><it>s </it></sup>mice are cross-bred with other mouse models of neurodegenerative disease in order to understand the neuroprotective processes conferred by the <it>Wld</it><sup><it>s </it></sup>mutation.</p>http://www.molecularneurodegeneration.com/content/2/1/21 |
| spellingShingle | Coleman Michael P Shipston Michael J Chen Philip E MacDonald Stephen HF Wishart Thomas M Gillingwater Thomas H Ribchester Richard R Design of a novel quantitative PCR (QPCR)-based protocol for genotyping mice carrying the neuroprotective Wallerian degeneration slow (<it>Wld</it><sup><it>s</it></sup>) gene Molecular Neurodegeneration |
| title | Design of a novel quantitative PCR (QPCR)-based protocol for genotyping mice carrying the neuroprotective Wallerian degeneration slow (<it>Wld</it><sup><it>s</it></sup>) gene |
| title_full | Design of a novel quantitative PCR (QPCR)-based protocol for genotyping mice carrying the neuroprotective Wallerian degeneration slow (<it>Wld</it><sup><it>s</it></sup>) gene |
| title_fullStr | Design of a novel quantitative PCR (QPCR)-based protocol for genotyping mice carrying the neuroprotective Wallerian degeneration slow (<it>Wld</it><sup><it>s</it></sup>) gene |
| title_full_unstemmed | Design of a novel quantitative PCR (QPCR)-based protocol for genotyping mice carrying the neuroprotective Wallerian degeneration slow (<it>Wld</it><sup><it>s</it></sup>) gene |
| title_short | Design of a novel quantitative PCR (QPCR)-based protocol for genotyping mice carrying the neuroprotective Wallerian degeneration slow (<it>Wld</it><sup><it>s</it></sup>) gene |
| title_sort | design of a novel quantitative pcr qpcr based protocol for genotyping mice carrying the neuroprotective wallerian degeneration slow it wld it sup it s it sup gene |
| url | http://www.molecularneurodegeneration.com/content/2/1/21 |
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