Strain Name:
FVB/N-Tg(Ckm-Prkaa2*D157A)1Ljg/Mmjax
Stock Number:
041177-JAX
Citation ID:
RRID:MMRRC_041177-JAX
Other Names:
AMPK alpha-2i TG

Strain Information

Ckm
Name: creatine kinase, muscle
Synonyms: M-CK, MCK, Ckmm
Type: Gene
Species: Mouse
Chromosome: 7
NCBI: 12715
HGNC: HGNC:1994
Homologene: 20432
Prkaa2
Name: protein kinase, AMP-activated, alpha 2 catalytic subunit
Type: Gene
Species: Rattus norvegicus (norway rat)
Chromosome: 5
Tg(Ckm-Prkaa2*D157A)1Ljg
Name: transgene insertion 1, Laurie J Goodyear
Synonyms: alpha2i TG, AMPKalpha2i TG
Type: Transgene
Species: Multi-species
Chromosome: unknown
Alteration at locus: Transgenic
Genetic Alterations
To render the catalytic subunit inactive, the aspartic acid at amino acid residue 157 of rat AMPK alpha 2 subunit was substituted to alanine. The AMPK alpha 2 subunit is tagged at the amino terminus with a hemagglutinin epitope. The recombinant DNA is driven by a muscle creatine kinase promoter.

HGVS:
  • Genbank RefSeq - mRNA: NM_178143.2
  • Genbank RefSeq, protein: NP_835279.2
  • Variant, nucleic acid level: c.470A>C
  • Variant, amino acid level, predicted: p.Asp157Ala
    Check this variant: LUMC Mutalyzer
Phenotype

Homozygous: Same as hemizygous

Hemizygous: In AMPK alpha2i TG mice basal AMPKalpha2 activity was barely detectable, whereas basal AMPK alpha1 activity was only partially reduced. Known AMPK stimuli including 5-aminoimidazole-4-carboxamide-1-beta-4-ribofuranoside (AICAR), rotenone (a Complex I inhibitor), dinitrophenol (a mitochondrial uncoupler), muscle contraction,and sorbitol (producing hyperosmolar shock) did not increase AMPK alpha2 activity in alpha2i TG mice, whereas 1 activation was attenuated by only 30-50%. AICAR- and rotenone-stimulated glucose transport was fully inhibited in AMPK alpha-2iTG mice; however, the lack of AMPK alpha-2 activity had no effect on contraction- or sorbitol-induced glucose transport in muscle. Mice breed and survive normally.

MeSH Terms
  • AMP-Activated Protein Kinases
  • Aminoimidazole Carboxamide/analogs & derivatives
  • Aminoimidazole Carboxamide/pharmacology
  • Animals
  • Biological Transport, Active/drug effects
  • Glucose/metabolism
  • In Vitro Techniques
  • Mice
  • Mice, Transgenic
  • Multienzyme Complexes/chemistry
  • Multienzyme Complexes/genetics
  • Multienzyme Complexes/metabolism
  • Muscle Contraction
  • Muscle, Skeletal/drug effects
  • Muscle, Skeletal/metabolism
  • Mutagenesis, Site-Directed
  • Osmolar Concentration
  • Protein-Serine-Threonine Kinases/chemistry
  • Protein-Serine-Threonine Kinases/genetics
  • Protein-Serine-Threonine Kinases/metabolism
  • Rats
  • Recombinant Proteins/chemistry
  • Recombinant Proteins/genetics
  • Recombinant Proteins/metabolism
  • Ribonucleotides/pharmacology
  • Rotenone/pharmacology
  • Signal Transduction
  • Sorbitol/pharmacology
  • AMP-Activated Protein Kinases/deficiency
  • AMP-Activated Protein Kinases/genetics
  • AMP-Activated Protein Kinases/metabolism
  • Biological Transport, Active
  • Cells, Cultured
  • Mice, Inbred C57BL
  • Mice, Inbred ICR
  • Mice, Knockout
  • Muscle Contraction/physiology
  • Muscle, Skeletal/physiology
  • Phosphatidylinositol 3-Kinases/metabolism
  • Proto-Oncogene Proteins c-akt/metabolism
  • Reactive Oxygen Species/metabolism
  • Stress, Mechanical
  • p38 Mitogen-Activated Protein Kinases/metabolism
  • Adenylate Kinase/metabolism
  • Glucose Transporter Type 4/metabolism
  • Green Fluorescent Proteins/metabolism
  • Kinetics
  • Male
  • Microtubules/metabolism
  • Muscle Fibers, Skeletal/physiology
  • Plasmids
  • Protein Transport/physiology
  • Sarcolemma/metabolism
  • Transfection
  • Antibodies, Phospho-Specific/pharmacology
  • Consensus Sequence
  • Dietary Fats/pharmacology
  • GTPase-Activating Proteins
  • Insulin/pharmacology
  • Muscle Contraction/drug effects
  • Muscle, Skeletal/enzymology
  • Mutant Proteins/metabolism
  • Mutation/genetics
  • Nuclear Proteins/genetics
  • Nuclear Proteins/metabolism
  • Phosphorylation/drug effects
  • Phosphoserine/metabolism
  • Blood Glucose/metabolism
  • Body Weight/drug effects
  • Dietary Fats/administration & dosage
  • Fatty Acids, Nonesterified/blood
  • Glucose Tolerance Test
  • Glycogen/metabolism
  • Immunoblotting
  • Insulin/blood
  • Insulin Resistance
  • Muscles/drug effects
  • Muscles/metabolism
  • Triglycerides/blood
  • Amino Acid Sequence
  • Cell Line
  • Enzyme Activation/drug effects
  • Molecular Sequence Data
  • Mutation
  • Myoblasts/cytology
  • Myoblasts/drug effects
  • Myoblasts/metabolism
  • Rats, Sprague-Dawley
  • p38 Mitogen-Activated Protein Kinases/genetics
  • Adaptation, Biological
  • Biomarkers/metabolism
  • Gene Expression Regulation, Enzymologic
  • Hexokinase/metabolism
  • Mitochondrial Proteins/metabolism
  • Myosin Heavy Chains/metabolism
  • Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha
  • Physical Conditioning, Animal
  • Protein Subunits/genetics
  • Protein Subunits/metabolism
  • Trans-Activators/metabolism
  • Transcription Factors
  • Amino Acid Substitution
  • Biological Transport
  • Female
  • Insulin/physiology
  • Models, Animal
  • Protein Kinases/genetics
  • Protein Kinases/metabolism
  • Calcium-Calmodulin-Dependent Protein Kinase Kinase
  • Calcium-Calmodulin-Dependent Protein Kinase Type 1
  • Calcium-Calmodulin-Dependent Protein Kinases/metabolism
  • Deoxyglucose/metabolism
  • Genetic Vectors
  • Humans
  • Phosphorylation
  • GTPase-Activating Proteins/drug effects
  • GTPase-Activating Proteins/metabolism
  • Protein Transport
  • Signal Transduction/physiology
  • Echocardiography
  • Heart/physiology
  • Myocardium/enzymology
Strain Development
To render the catalytic subunit inactive, the aspartic acid at amino acid residue 157 of rat AMPK 2 subunit (Prkaa2) was substituted to alanine by a PCR-based site direct mutagenesis. Complementary primers spanning the residue to be mutated were synthesized and used for the mutagenesis. Mice expressing the inactive alpha2 tagged at the amino terminus with a hemagglutinin epitope (alpha 2i TG mice) were generated by injecting the recombinant DNA driven by a muscle creatine kinase promoter into fertilized FVB/NTac mouse oocytes. FVB/NTac founders were bred with FVB/NTac mice.
Suggested Control Mice
Wild-type littermates
MMRRC Genetic QC Summary
The MMRRC Centers have developed a genetic QC pipeline using MiniMUGA array genotyping to provide additional information on strain backgrounds for MMRRC congenic and inbred strains. For more information on when data may be available, or to request genotyping for a strain of interest, please contact csmmrrc@jax.org. Older strains may not have this information.
  • Diabetes
  • Metabolism
  • Obesity
  • Research Tools
Donor
Laurie Goodyear, Ph.D., Joslin Diabetes Center.
Primary Reference
  • Fujii N, Hirshman MF, Kane EM, Ho RC, Peter LE, Seifert MM, Goodyear LJ. AMP-activated protein kinase alpha2 activity is not essential for contraction- and hyperosmolarity-induced glucose transport in skeletal muscle. J Biol Chem. 2005 Nov 5;280(47):39033-41. Epub 2005 Sep 26. (Medline PMID: 16186119)
  • Chambers MA, Moylan JS, Smith JD, Goodyear LJ, Reid MB. Stretch-stimulated glucose uptake in skeletal muscle is mediated by reactive oxygen species and p38 MAP-kinase. J Physiol. 2009 Jul 1;587(Pt 13):3363-73. Epub 2009 Apr 29. (Medline PMID: 19403598)
  • Lauritzen HP, Galbo H, Toyoda T, Goodyear LJ. Kinetics of contraction-induced GLUT4 translocation in skeletal muscle fibers from living mice. Diabetes. 2010 Sep;59(9):2134-44. Epub 2010 Jul 9. (Medline PMID: 20622170)
  • Vichaiwong K, Purohit S, An D, Toyoda T, Jessen N, Hirshman MF, Goodyear LJ. Contraction regulates site-specific phosphorylation of TBC1D1 in skeletal muscle. Biochem J. 2010 Oct 15;431(2):311-20. (Medline PMID: 20701589)
  • Fujii N, Ho RC, Manabe Y, Jessen N, Toyoda T, Holland WL, Summers SA, Hirshman MF, Goodyear LJ. Ablation of AMP-activated protein kinase alpha2 activity exacerbates insulin resistance induced by high-fat feeding of mice. Diabetes. 2008 Nov;57(11):2958-66. Epub 2008 Aug 26. (Medline PMID: 18728234)
  • Ho RC, Fujii N, Witters LA, Hirshman MF, Goodyear LJ. Dissociation of AMP-activated protein kinase and p38 mitogen-activated protein kinase signaling in skeletal muscle. Biochem Biophys Res Commun. 2007 Oct 19;362(2):354-9. Epub 2007 Aug 7. (Medline PMID: 17709097)
  • R ckl KS, Hirshman MF, Brandauer J, Fujii N, Witters LA, Goodyear LJ. Skeletal muscle adaptation to exercise training: AMP-activated protein kinase mediates muscle fiber type shift. Diabetes. 2007 Aug;56(8):2062-9. Epub 2007 May 18. (Medline PMID: 17513699)
  • Fujii N, Seifert MM, Kane EM, Peter LE, Ho RC, Winstead S, Hirshman MF, Goodyear LJ. Role of AMP-activated protein kinase in exercise capacity, whole body glucose homeostasis, and glucose transport in skeletal muscle -insight from analysis of a transgenic mouse model-. Diabetes Res Clin Pract. 2007 Sep;77 Suppl 1:S92-8. Epub 2007 Apr 23. (Medline PMID: 17452058)
  • Witczak CA, Fujii N, Hirshman MF, Goodyear LJ. Ca2+/calmodulin-dependent protein kinase kinase-alpha regulates skeletal muscle glucose uptake independent of AMP-activated protein kinase and Akt activation. Diabetes. 2007 May;56(5):1403-9. Epub 2007 Feb 7. (Medline PMID: 17287469)
  • Kramer HF, Witczak CA, Fujii N, Jessen N, Taylor EB, Arnolds DE, Sakamoto K, Hirshman MF, Goodyear LJ. Distinct signals regulate AS160 phosphorylation in response to insulin, AICAR, and ontraction in mouse skeletal muscle. Diabetes. 2006 Jul;55(7):2067-76. (Medline PMID: 16804077)
  • Musi N, Hirshman MF, Arad M, Xing Y, Fujii N, Pomerleau J, Ahmad F, Berul CI, Seidman JG, Tian R, Goodyear LJ. Functional role of AMP-activated protein kinase in the heart during exercise. FEBS Lett. 2005 Apr 11;579(10):2045-50. (Medline PMID: 15811316)

Colony and Husbandry Information

Cryo-recovered strains distributed by the MMRRC at JAX are shipped to the customer from the Pathogen & Opportunistic-Free Animal Room G200 - see https://www.jax.org/jax-mice-and-services/customer-support/customer-service/animal-health/health-status-reports.

Mice recovered from a cryo-archive will have health surveillance performed on recipient females. Health reports will be provided prior to shipment. If you require additional health status information, please email csmmrrc@jax.org.
Coat Color
White
Other
They look like wild-type FVB/NTac mice
MMRRC Breeding System
Other or uncertain
Generation
N10+ (FVB/NTac), F10+
Overall Breeding Performance
Excellent
NOTE: "Hemizygote" as used here refers to males carrying a mutation on the X Chromosome or mice of either sex carrying an inserted transgene with no homologous allele on the other chromosome.
Viability and Fertility: Female Male Comments
Homozygotes are viable: Yes Yes
Homozygotes are fertile: Yes Yes
Hetero/Hemizygotes are fertile: Yes Yes
Age Reproductive Decline: 6 to 8 months 8 to 12 months
Average litter size
5-9
Recommended wean age
3-4 weeks
Average Pups Weaned
5-9

Order Request Information

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 csmmrrc@jax.org for more information.

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.

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.

Click button to Request this one strain. (Use the MMRRC Catalog Search to request more than one strain.)
MMRRC Item # Description Distribution Fee / Unit (US $)
*Shipping & Handling not included*
Units Notes
041177-JAX-SPERM Cryo-preserved spermatozoa $437.00 / $437.00
Non-Profit / For-Profit
Aliquot Approximate quantity3
041177-JAX-RESUS Litter recovered from cryo-archive $2,022.00 / $2,022.00
Non-Profit / For-Profit
Litter Recovered litter4; additional fees for any special requests.
Cryopreserved material may be available upon request, please inquire to csmmrrc@jax.org for more information.

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.

To request material from the MMRRC: Please fill out our on-line request form (accessible from the catalog search results page, or click the Request this Strain button in the fees section). If you have questions or need assistance completing this form, you may call Customer Service at (800) 910-2291 (in USA or Canada) or (530) 757-5710 (international calls). Before you call, please have with you: the MMRRC item number, quantity needed, Bill-to and Ship-to contact information.