Strain Detail Sheet

Strain Name    :

C57BL/6J-Map3k8m1Btlr/Mmucd

Stock Number :

030499-UCD

Other Names   :

Sluggish, C57BL/6J-Map3k8m1Btlr/Mmcd

Gene Information

Gene Details [Including genotyping protocols]

(provided by MGI)
Allele Symbol: Map3k8M1Btlr
Name: mutation 1, Bruce Beutler
Alteration at locus: Chemically Induced
Gene Symbol: Map3k8
Name: mitogen-activated protein kinase kinase kinase 8
Chromosome: 18
Alteration at locus: Chemically Induced

Genetic Alterations:
The Sluggish mutation was mapped to Chromosome 18, and is a T to A transversion in the acceptor splice site of intron 5 at position 13346 of the Map3k8 gene (Genbank genomic region NC_000084 for linear genomic DNA sequence of Map3k8).

Genotype Determination:

ES Cell Line: Not Applicable

Strain Description [Including phenotype, strain background, strain development and suggested control mice]

Phenotype

Homozygous phenotype: The Sluggish phenotype was identified in a screen of homozygous ENU-induced G3 mutant mice for altered responses to Toll-like receptor (TLR) ligands. Peritoneal macrophages from Sluggish homozygotes did not produce tumor necrosis factor (TNF)-α in response to the TLR2/6 ligands MALP2 (macrophage-activating lipopeptide 2) and peptidoglycan, or to the TLR9 ligand CpG DNA. These macrophages also exhibited a partially reduced TNF-α response to other TLR ligands, such as the TLR4 ligand lipopolysaccharide (LPS), the TLR1/2 ligand Pam3CSK4 (a triacyl lipopeptide), the TLR7 ligand resiquimod (a ssRNA mimetic), and the TLR3 ligand poly I:C (a dsRNA mimetic). The TNF-α response to TLR7 is greatly reduced, while the responses to TLR4, TLR1/2, and TLR3 are intermediate. In addition, the interferon (IFN) α/β response to TLR7 and TLR9 signaling was abolished as was the production of IL- 1β induced by LPS, but the interferon response to LPS and poly I:C was normal. The defect in the TNF-α response to TLR signaling is likely to be one of post-transcriptional proteolytic processing as normal levels of intracellular TNF-α are observed in MALP2, peptidoglycan, and CpG stimulated Sluggish macrophages. Peritoneal macrophages from Sluggish homozygotes showed normal resistance to mouse cytomegalovirus (MCMV) and normal restriction of GFP expression when infected by a non- replicating recombinant Adenovirus 5 vector, but failed to produce TNF-α in response to viral infection. Sluggish homozygous mice have impaired natural killer (NK) cell cyotoxicity in vivo as measured by reduced clearance of MHC class 1-deficient cells.

A quarter of the expected number of heterozygous and very few homozygous Sluggish animals are born, suggesting that the Sluggish mutation affects embryonic survival. The stage at which death occurs is unknown. Surviving Sluggish homozygous animals exhibit no visible phenotypic defects, are viable, and can breed.

Heterozygous phenotype: Heterozygous Sluggish animals displayed a partial TNF-α response to TLR2/6 signaling and reduced numbers survive birth.


Mammalian Phenotype Terms:(provided by MGI)      Extend all MPTs
      assigned by genotype
Map3k8m1Btlr/Map3k8+
        C57BL/6J-Map3k8<m1Btlr>
  • mortality/aging
    • partial prenatal lethality (MGI Ref ID J:140986)
      • only ~25% of the expected number of heterozygous mutant mice are born; the developmental stage at which death occurs has not been determined
      • surviving heterozygotes exhibit no overt phenotypic defects, are viable, and can breed
  • immune system phenotype
    • abnormal tumor necrosis factor secretion (MGI Ref ID J:140986)
      • heterozygous macrophages secrete somewhat diminished levels of TNF-alpha in response to lower doses of the TLR1/2 ligand Pam3SK4, but slightly elevated levels in response to higher doses
      • decreased tumor necrosis factor secretion (MGI Ref ID J:140986)
        • peritoneal macrophages from mice heterozygous for this mutation exhibit impairment in TNF-alpha secretion in response to MALP2 and PGN signaling via TLR1/2, CpG-DNA signaling via TLR9, and LPS signaling via TLR4
        • resiquimod signaling via TLR7 stimulates statistically normal TNF-alpha secretion by heterozygous mutant macrophages
Map3k8m1Btlr/Map3k8m1Btlr
        C57BL/6J-Map3k8<m1Btlr>
  • mortality/aging
    • partial prenatal lethality (MGI Ref ID J:140986)
      • very few mice homozygous for this mutation are born; the developmental stage at which death occurs has not been determined
      • surviving homozygous mutants exhibit no overt phenotypic defects, are viable, and can breed
  • immune system phenotype
    • abnorma l response to infection (MGI Ref ID J:140986)
      • peritoneal macrophages from homozygous mutants fail to produce TNF-alpha in response to viral infection
      • homozygous mutant macrophages show normal resistance to mouse cytomegalovirus (MCMV) and normal restriction of GFP expression when infected by a non-replicating recombinant Adenovirus 5 vector
    • decreased interferon-alpha secretion (MGI Ref ID J:140986)
      • peritoneal macrophages from homozygous mutant mice fail to produce interferon alpha/beta (IFN-alpha/beta) in response to signaling via TLR7 or TLR9
      • mutant macrophages exhibit a normal interferon response to LPS (via TLR4) and to poly I:C (via TLR3)
    • decreased interferon-beta secretion (MGI Ref ID J:140986)
      • peritoneal macrophages from homozygous mutant mice fail to produce interferon alpha/beta (IFN-alpha/beta) in response to signaling via TLR7 or TLR9
      • mutant macrophages exhibit a normal interferon response to LPS (via TLR4) and to poly I:C (via TLR3)
    • decreased interleukin-1 beta secretion (MGI Ref ID J:140986)
      • the interleukin 1 beta (IL-1beta) response to LPS signaling via TLR4 is abolished in mutant macrophages
    • decreased interleukin-6 secretion (MGI Ref ID J:140986)
      • mutant macrophages have an impaired interleukin- (IL-) 6 response to PGN (via TLR2/6), to Pam3CSK4 (via TLR1/2), and to resiquimod (via TLR7)
    • decreased tumor necrosis factor secretion (MGI Ref ID J:140986)
      • peritoneal macrophages from homozygous mutants fail to produce TNF-alpha in response to viral infection
      • peritoneal macrophages from homozygous mutants mice do not secrete tumor necrosis factor (TNF) -alpha in response to the TLR2/6 ligands MALP2 (macrophage-activating lipopeptide 2) and peptidoglycan or to CpG DNA, a TLR9 ligand
      • although it is not secreted, normal intracellular levels of TNF-alpha are observed in MALP2-, peptidoglycan-, and CpG-stimulated mutant macrophages
      • secretion of TNF-alpha in response to the TLR4 ligand, lipopolysaccharide (LPS), and to resiquimod, a ssRNA analog that signals via TLR7, is greatly reduced
      • homozygous mutant macrophages exhibit a partially attenuated TNF-alpha response to the TLR1/2 ligand Pam3CSK4, a triacyl lipopeptide; and to the TLR3 ligand poly I:C, a double-stranded DNA mimetic
    • impaired NK cell cytolysis (MGI Ref ID J:140986)
      • homozygous muta nt mice have impaired natural killer (NK) cell cyotoxicity in vivo as measured by reduced clearance of MHC class 1-deficient cells
    • increased interleukin-12b secretion (MGI Ref ID J:140986)
      • production of interleukin- (IL-) 12/p40 by mutant macrophages is elevated in response to signaling via TLR7 and vai TLR9

Strain of Origin: C57BL/6J

Strain genetic background: C57BL/6J

Strain Development: The sluggish mutation was induced by ENU mutagenesis on the C57BL/6J background, and was discovered in G3 animals. All subsequent crosses to maintain strain were on a C57BL/6J background.

Suggested Control Mice:

  • Wildtype littermates

Research Applications

  • Cancer
  • Cell Biology
  • Immunology and Inflammation

Strain Origin

Donor: Bruce Beutler, M.D., The Scripps Research Institute

Primary Reference:

  • Xiao N; Eidenschenk C; Krebs P; Brandl K; Blasius AL; Xia Y; Khovananth K; Smart NG; Beutler B, The Tpl2 mutation Sluggish impairs type I IFN production and increases susceptibility to group B streptococcal disease., J Immunol 2009 Dec 15;183(12):7975-83 (Medline PMID: 19923465)
  • For additional information see: Mutagenetix, a catalog of mutations identified in the Beutler Laboratory at The Scripps Research Institute.)

Colony and Husbandry Information

Special Considerations

None

Health Status Report

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 mmrrc@ucdavis.edu.

Order Request Information

Availability Level

Limited quantities of breeder mice (recovered litter) are available from a cryoarchive; recovered litter usually available to ship in 3 to 4 months.

Conditions of Distribution [Including applicable technology transfer agreements]

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.

The donor or their institution limits the distribution to non-profit institutions only.

Fees

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 $)
Units Notes
030499-UCD-RESUSLitter recovered from cryo-archive
$2,022.00
Non-Profit
Litter Recovered litter1; additional fees for any special requests.
030499-UCD-SPERMCryo-preserved spermatozoa
$437.00
Non-Profit
Aliquot Approximate quantity.2
030499-UCD-EMBRYOCryo-preserved embryos
$1,038.00
Non-Profit
Aliquot Approximate quantity3: 20-40 embryos / aliquot

1 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.

2 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.

3 An aliquot contains a sufficient number of embryos (in one or more vials and based on the transfer success rate of the MMRRC facility) to transfer to at least two recipients. The MMRRC makes no guarantee concerning embryo transfer success experienced in the recipient investigator's laboratory. Neither gender nor genotype ratios are guaranteed.

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 above). 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.



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.



The MMRRC is a collaborative effort, funded by grants from DPCPSI of the NIH.

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