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Mapping of the spontaneous deletion in the Ap3d1 gene of mocha mice: fast and reliable genotyping
BACKGROUND: The mocha mouse carries a spontaneous deletion in the Ap3d1 gene, encoding the delta 1 subunit of the adaptor related protein complex 3, (Ap3d1), and subsequently lack the expression of functional AP-3. This leads to a deficiency in vesicle transport and storage, which affects neurotrans...
Autores principales: | , , , |
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Formato: | Texto |
Lenguaje: | English |
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BioMed Central
2008
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2612676/ https://www.ncbi.nlm.nih.gov/pubmed/19032734 http://dx.doi.org/10.1186/1756-0500-1-119 |
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author | Drasbek, Kim Ryun Holm, Mai Marie Delenclos, Marion Jensen, Kimmo |
author_facet | Drasbek, Kim Ryun Holm, Mai Marie Delenclos, Marion Jensen, Kimmo |
author_sort | Drasbek, Kim Ryun |
collection | PubMed |
description | BACKGROUND: The mocha mouse carries a spontaneous deletion in the Ap3d1 gene, encoding the delta 1 subunit of the adaptor related protein complex 3, (Ap3d1), and subsequently lack the expression of functional AP-3. This leads to a deficiency in vesicle transport and storage, which affects neurotransmitter vesicle turnover and release in the central nervous system. Since the genomic sequence of the Ap3d1 gene of mocha mouse is not known, precise mapping of the deletion as well as reliable genotyping protocols are lacking. FINDINGS: We sequenced the Ap3d1 gene (HGNC GeneID: 8943) around the deletion site in the mocha mouse and revealed a 10639 bp deletion covering exon 2 to 6. Subsequently, new PCR primers were designed yielding a reliable genotyping protocol of both newborn and adult tissue. To examine the genotypes further, hippocampal neurons were cultured from mocha and control mice. Patch-clamp recordings showed that mocha neurons had a higher input resistance, and that autaptic EPSC in mocha cultures depressed faster and stronger as compared with control cultures. CONCLUSION: Our study reports the sequence of the deleted part of the Ap3d1 gene in mocha mice, as well as a reliable PCR-based genotyping protocol. We cultured hippocampal neurons from control and mocha mice, and found a difference in input resistance of the neurons, and in the synaptic short-term plasticity of glutamatergic autapses showing a larger synaptic depression than controls. The described procedures may be useful for the future utilization of the mocha mouse as a model of defective vesicle biogenesis. Importantly, as genotyping by eye color is complicated in newborn mice, the designed protocol is so fast and reliable that newborn mice could rapidly be genotyped and hippocampal neurons dissociated and cultured, which is normally best done at P0-P2. |
format | Text |
id | pubmed-2612676 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2008 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-26126762008-12-31 Mapping of the spontaneous deletion in the Ap3d1 gene of mocha mice: fast and reliable genotyping Drasbek, Kim Ryun Holm, Mai Marie Delenclos, Marion Jensen, Kimmo BMC Res Notes Short Report BACKGROUND: The mocha mouse carries a spontaneous deletion in the Ap3d1 gene, encoding the delta 1 subunit of the adaptor related protein complex 3, (Ap3d1), and subsequently lack the expression of functional AP-3. This leads to a deficiency in vesicle transport and storage, which affects neurotransmitter vesicle turnover and release in the central nervous system. Since the genomic sequence of the Ap3d1 gene of mocha mouse is not known, precise mapping of the deletion as well as reliable genotyping protocols are lacking. FINDINGS: We sequenced the Ap3d1 gene (HGNC GeneID: 8943) around the deletion site in the mocha mouse and revealed a 10639 bp deletion covering exon 2 to 6. Subsequently, new PCR primers were designed yielding a reliable genotyping protocol of both newborn and adult tissue. To examine the genotypes further, hippocampal neurons were cultured from mocha and control mice. Patch-clamp recordings showed that mocha neurons had a higher input resistance, and that autaptic EPSC in mocha cultures depressed faster and stronger as compared with control cultures. CONCLUSION: Our study reports the sequence of the deleted part of the Ap3d1 gene in mocha mice, as well as a reliable PCR-based genotyping protocol. We cultured hippocampal neurons from control and mocha mice, and found a difference in input resistance of the neurons, and in the synaptic short-term plasticity of glutamatergic autapses showing a larger synaptic depression than controls. The described procedures may be useful for the future utilization of the mocha mouse as a model of defective vesicle biogenesis. Importantly, as genotyping by eye color is complicated in newborn mice, the designed protocol is so fast and reliable that newborn mice could rapidly be genotyped and hippocampal neurons dissociated and cultured, which is normally best done at P0-P2. BioMed Central 2008-11-25 /pmc/articles/PMC2612676/ /pubmed/19032734 http://dx.doi.org/10.1186/1756-0500-1-119 Text en Copyright © 2008 Holm et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( (http://creativecommons.org/licenses/by/2.0) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Short Report Drasbek, Kim Ryun Holm, Mai Marie Delenclos, Marion Jensen, Kimmo Mapping of the spontaneous deletion in the Ap3d1 gene of mocha mice: fast and reliable genotyping |
title | Mapping of the spontaneous deletion in the Ap3d1 gene of mocha mice: fast and reliable genotyping |
title_full | Mapping of the spontaneous deletion in the Ap3d1 gene of mocha mice: fast and reliable genotyping |
title_fullStr | Mapping of the spontaneous deletion in the Ap3d1 gene of mocha mice: fast and reliable genotyping |
title_full_unstemmed | Mapping of the spontaneous deletion in the Ap3d1 gene of mocha mice: fast and reliable genotyping |
title_short | Mapping of the spontaneous deletion in the Ap3d1 gene of mocha mice: fast and reliable genotyping |
title_sort | mapping of the spontaneous deletion in the ap3d1 gene of mocha mice: fast and reliable genotyping |
topic | Short Report |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2612676/ https://www.ncbi.nlm.nih.gov/pubmed/19032734 http://dx.doi.org/10.1186/1756-0500-1-119 |
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