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Two amino acid changes were engineered into the Atp1a2 gene rendering the Na,K-ATPase α2 isoform resistant to ouabain binding. In wild type mice, this isoform is sensitive to ouabain binding, with nanomolar affinity.
A targeting vector designed to alter several amino acids impacting ouabain binding was transfected into Duffy embryonic stem (ES) cells (derived from 129S6). Correctly targeted ES cells were injected into blastocysts. Chimeric offspring were bred to Black Swiss mice for two generations. The mice were further backcrossed to B6 mice for and extended period of time.
Dostanic I, Lorenz JN, Schultz Jel J, Grupp IL, Neumann JC, Wani MA, Lingrel JB. The alpha2 isoform of Na,K-ATPase mediates ouabain-induced cardiac inotropy in mice. J Biol Chem. 2003 Dec 26;278(52):53026-34. doi: 10.1074/jbc.M308547200. Epub 2003 Oct 14. (Medline PMID: 14559919)
Radzyukevich TL, Neumann JC, Rindler TN, Oshiro N, Goldhamer DJ, Lingrel JB, Heiny JA. Tissue-specific role of the Na,K-ATPase α2 isozyme in skeletal muscle. J Biol Chem. 2013 Jan 11;288(2):1226-37. doi: 10.1074/jbc.M112.424663. Epub 2012 Nov 28. (Medline PMID: 23192345)
Dostanic I, Schultz Jel J, Lorenz JN, Lingrel JB. The alpha 1 isoform of Na,K-ATPase regulates cardiac contractility and functionally interacts and co-localizes with the Na/Ca exchanger in heart. J Biol Chem. 2004 Dec 24;279(52):54053-61. doi: 10.1074/jbc.M410737200. Epub 2004 Oct 14. (Medline PMID: 15485817)
Bossuyt J, Despa S, Han F, Hou Z, Robia SL, Lingrel JB, Bers DM. Isoform specificity of the Na/K-ATPase association and regulation by phospholemman. J Biol Chem. 2009 Sep 25;284(39):26749-57. doi: 10.1074/jbc.M109.047357. Epub 2009 Jul 28. (Medline PMID: 19638348)
Chomczynski PW, Vires KM, Rymaszewski M, Heiny JA. A real-time PCR method to genotype mutant mouse models with altered affinity for cardiotonic steroids on the Na,K-ATPase. PLoS One. 2022 Apr 21;17(4):e0267348. doi: 10.1371/journal.pone.0267348. (Medline PMID: 35446892)
Despa S, Lingrel JB, Bers DM. Na(+)/K)+)-ATPase α2-isoform preferentially modulates Ca2(+) transients and sarcoplasmic reticulum Ca2(+) release in cardiac myocytes. Cardiovasc Res. 2012 Sep 1;95(4):480-6. doi: 10.1093/cvr/cvs213. Epub 2012 Jun 27. (Medline PMID: 22739122)
Colony Surveillance Program and Current Health Reports
Limited quantities of breeder mice (recovered litter) are available from a cryoarchive; recovered litter usually available to ship in 3 to 4 months.
Cryopreserved material may be available upon request, please inquire to mmrrc@med.unc.edu for more information.
A Commercial License Agreement from the Donor is required for for-profit entities to use this strain. For more information, please contact Geoffrey Pinski.
Distribution of this strain requires submission of the MMRRC Conditions of Use (COU). A link to the COU web form will be provided via email after an order has been placed; the form should be completed then or the email forwarded to your institutional official for completion.
A Commercial License Agreement from the Donor is required for for-profit entities to use this strain. For more information, please contact Geoffrey Pinski
Additional charges may apply for any special requests. Shipping costs are in addition to the basic distribution/resuscitation fees. Information on shipping costs and any additional charges will be provided by the supplying MMRRC facility.
1 The distribution fee covers the expense of rederiving mice from a live mouse; you will receive the resulting litter. The litter will contain at minimum one mutant carrier; the actual number of animals and the gender and genotype ratios will vary. (Typically, multiple breeder pairs can be established from the recovered litter.) Prior to shipment, the MMRRC will provide information about the animals recovered. If you anticipate or find that you need to request specific genotypes, genders or quantities of mice in excess of what is likely from a resuscitated litter, you may discuss available options and pricing with the supplying MMRRC facility.
2 An aliquot contains a sufficient number of embryos (in one or more vials or straws and based on the transfer success rate of the MMRRC facility) to transfer into one to three recipients. The MMRRC makes no guarantee concerning embryo transfer success experienced in the recipient investigator's laboratory. Neither gender nor genotype ratios are guaranteed.
3 An aliquot is one straw or vial with sufficient sperm to recover at least one litter of mice, as per provided protocols, when performed at the MMRRC facility. The MMRRC makes no guarantee concerning the success of these procedures when performed outside the MMRRC facilities.
4 The distribution fee covers the expense of resuscitating mice from the cryo-archive; you will receive the resulting litter. The litter will contain at minimum one mutant carrier; the actual number of animals and the gender and genotype ratios will vary. (Typically, multiple breeder pairs can be established from the recovered litter.) Prior to shipment, the MMRRC will provide information about the animals recovered. If you anticipate or find that you need to request specific genotypes, genders or quantities of mice in excess of what is likely from a resuscitated litter, you may discuss available options and pricing with the supplying MMRRC facility.