Overview

In this strain a PGK-neo cassette replaces sequence including exon 2 of the Lum (lumican) gene. These mice exhibit corneal opacity, poor wound healing, are hyporesponsive to LPS-induced sepsis and may be useful in studies of corneal dystrophy, extracellular matrix and neutrophil migration during the immune response.

Donating Investigator

Shukti Chakravarti, Johns Hopkins University

Genetic overview

Genetic Background	Generation

Lum^tm1Chak

Allele Type	Gene Symbol	Gene Name
Targeted (Null/Knockout)	Lum	lumican

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Research Applications

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Base Price

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$2,854.50 Domestic price Cryo Recovery

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Details

Detailed Description

The protein encoded by the Lum (lumican) gene, lumican, is a member of the small leucine-rich proteoglycan family, the predominant corneal keratan sulfate proteoglycan, is also found in interstitial extracellular matrices. Lumican regulates collagen fibril structure and is involved with the innate immune response, wound healing and neutrophil migration. These mice carry a targeted mutation for the Lum gene in which exon 2 is replaced by a NEO cassette. Mice that are homozygous for the targeted mutation are viable and fertile, but homozygous female mice on the C57BL/6 background are not good breeders. Homozygotes on the C57BL/6J background have loose skin around the neck, and older mice develop spontaneous lesion and tears of the skin. No gene product (protein) is detected by Western blot analysis of eye tissue, bone marrow extracts and peritoneal lavage from homozygotes. Homozygotes exhibit hyperextensible (8-fold reduced tensile strength) skin with abnormal collagen fibril structure and develop corneal opacity. Histological analysis of homozygotes reveals disorganized disorganized collagen fibrils with increased diameter in connective tissues of the skin, tendon, and cornea. Polymorphonuclear neutrophils from homozygotes exhibit poor chemotactic migration. In cell culture assays peritoneal macrophages (from homozygotes) exhibit defective phagocytosis of gram-negative bacteria. In response to LPS challenge, peritoneal macrophages and polymorphonuclear neutrophils (from homozygotes) exhibit a sub-normal production of proinflammatory cytokine (TNFalpha and IL6) production. Mutant mice are hyporesponsive to LPS-induced septic shock compared to wildtype controls and clearance of Gram-negative bacterial infections is impaired. During backcrossing, the Y chromosome may not have been fixed to the C57BL/6J genetic background.

Development

A targeting vector designed by Dr. Shukti Chakravarti, while at Case Western Reserve University, containing a PGK-neo cassette was used to disrupt 3kb of sequence, including exon 2. The construct was electroporated into 129S/Sv (129/Sv) derived CT129#25 embryonic stem (ES) cells. Correctly targeted ES cells were injected into CD-1 blastocysts. The resulting chimeric animals were tested for germline transmission by intercrossing. The donating investigator reported that these mice were backcrossed to C57BL/6J for 10 generations (see SNP note below). During backcrossing, the Y chromosome may not have been fixed to the C57BL/6J genetic background.
Upon arrival at The Jackson Laboratory, the mice were crossed to C57BL/6J (Stock No. 000664) at least once to establish the colony.

A 32 SNP (single nucleotide polymorphism) panel analysis, with 27 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. While the 27 markers throughout the genome suggested a C57BL/6 genetic background, 4 of 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a mixed C57BL/6J ; C57BL/6N genetic background.

Control Suggestions

Wild-type from the colony
000664 C57BL/6J

Additional Information

Considerations for Choosing Controls

Selected References

Chakravarti S; Magnuson T; Lass JH; Jepsen KJ; LaMantia C; Carroll H. 1998. Lumican regulates collagen fibril assembly: skin fragility and corneal opacity in the absence of lumican. J Cell Biol 141(5):1277-86PubMed: 9606218 MGI: J:48068

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Genetics

Lum^tm1Chak

Allele Symbol: Lum^tm1Chak

Allele Name	targeted mutation 1, Shukti Chakravarti
Allele Type	Targeted (Null/Knockout)
Allele Synonym(s)	Lum-; lum^tm1Sc
Gene Symbol and Name	Lum, lumican
Gene Synonym(s)
Strain of Origin	129S/Sv
Chromosome	10
General Note	ES cell line = CT 129#25.
Molecular Note	A neomycin resistance cassette replaced 3kb of sequence containing exon 2.

Disease/Phenotype

Disease Terms

Model with phenotypic similarity to human disease where etiologies are distinct. Human genes are associated with this disease. Orthologs of these genes do not appear in the mouse genotype(s).

Ehlers-Danlos syndrome classic type 1

Research Areas By Phenotype

This mouse can be used to support research in many areas including:

Mammalian Phenotype Terms by Genotype

The following phenotype information is associated with a similar, but not exact match to this JAX^® Mice strain

Genotype: Lum^tm1Chak/Lum^tm1Chak
involves: 129S/Sv * CD-1

cardiovascular system phenotype

bruising
- homozygotes bruise easily

integument phenotype

skin lesions
- homozygotes display an increased incidence of skin lesions

abnormal dermal layer morphology
- tail skin collagen fibrils are less affected relative to corneal collagen fibrils
- homozygotes display a disorganized and loosely arranged dermal connective tissue
- mutant dermal fibroblasts appear disoriented relative to wild-type
- in tail preparations (skin and tendon), dermal regions show increased interfibrillar spacing and a wide range of fiber diameter

loose skin
- homozygotes exhibit skin laxity and fragility

decreased skin tensile strength
- homozygotes display an 86% reduction in skin tensile strength relative to wild-type

mammalian phenotype

cardiovascular system phenotype
- Normal - homozygotes display no major cardiac valvular defects

digestive/alimentary phenotype
- Normal - homozygotes display no major gastrointestinal tract defects

muscle phenotype

abnormal tendon morphology
- at 1-3 months, mutant tendons display only slight alterations in collagen fibril diameter relative to wild-type; no differences are detected thereafter
- at P10, mutant tendons have relatively equal numbers of fibrils with intermediate diameters (between 65- and 140-nm)
- at P4, mutant tendons show a shift in the relatively homogeneous collagen fibril diameter distributions to larger diameters (72-nm versus 64-nm in wild-type)

skeleton phenotype

abnormal tendon morphology
- at P10, mutant tendons have relatively equal numbers of fibrils with intermediate diameters (between 65- and 140-nm)
- at P4, mutant tendons show a shift in the relatively homogeneous collagen fibril diameter distributions to larger diameters (72-nm versus 64-nm in wild-type)
- at 1-3 months, mutant tendons display only slight alterations in collagen fibril diameter relative to wild-type; no differences are detected thereafter

vision/eye phenotype

abnormal cornea morphology
- at both 2- and 6-months, X-ray patterns from mutant corneas fail to register any measurable subsidiary X-ray reflection, indicating a broader than normal range of fibril diameters
- mutant corneas display abnormal arrangement of collagen fibrils and keratinocytes
- at 6 months, the average collagen fibril spacing is marginally higher (~7%) in mutant corneas relative to wild-type
- X-ray diffraction patterns from mutant corneas revealed abnormally diffuse interfibrillar reflections
- TEM revealed that mutant corneal collagen fibrils show a wide range of fiber diameter (i.e. thicker fibrils in addition to fibrils of normal diameter), shape irregularities due to lateral fusions, and increased and non-uniform interfibrillar spacing

abnormal cornea posterior stroma morphology
- also, the posterior stroma displays abnormally large fibril aggregates, aberrant fibril packing, and impaired lamellar organization
- notably, the anterior stroma appears relatively unaffected
- the posterior stroma contains numerous large-diameter collagen fibrils, many with irregular contours, indicating abnormal lateral growth

abnormal corneal stroma development
- homozygotes display normal development of the corneal stroma between E13.5 and E16.5
- in contrast to wild-type corneal stroma, mutant stroma fails to undergo swelling from P8 to P12 (eyelid opening stage), followed by thinning at P14

corneal opacity
- whole eyes from mutant mice exhibit a 25% reduction in total ocular keratan sulfate levels, contributing to reduced corneal transparency
- at 5 to 34 weeks, 44 out of 51 homozygotes show bilateral corneal clouding with a peripheral ring-like clear zone
- also, corneal clouding is progressive: all homozygotes show an increase in opacity over time
- corneal opacity is age-dependent: by 8-12 weeks, most homozygotes have cloudy corneas
- importantly, no mutants exhibit total opacification where iris details are undetectable

sclera thinning
- in the sclera, the range and distribution of collagen fibril diameter is not severely affected
- mutant scleras display lamellar disorganization and occasional fibril-poor areas
- homozygotes show a significant reduction in posterior sclera thickness and a decrease in the number of lamellae

abnormal corneal stroma morphology
- the entire mutant stroma appears bright due to increased backscattered light
- beginning at 5 weeks, the mutant stroma appears uniformly hazy and opalescent

increased corneal light-scattering
- in contrast, corneas from mutant neonates show a significant increase in light-scattering at 3 weeks, when stroma thickness is reduced significantly
- on average, homozygotes show a 3-fold increase in backscattering of light, with maximal increase restricted to the posterior stroma
- in mutant posterior corneas, light-scattering is diffuse as opposed to granular, and remains elevated above wild-type levels, in the absence of significant changes in keratocyte proliferation or differentiation
- corneas from wild-type neonates show a rapid loss of light-scattering, decreasing by 50% from P1 to P12 (eyelid opening), concomittant with a 60% decrease in keratocyte density

decreased corneal stroma thickness
- in contrast to wild-type, mutant stroma never increases in thickness from week 1 to 2, remains markedly thinner at 2 weeks, and becomes significantly thinner from week 2 to week 3 after eyelid opening
- at this age, mutant mice also show a 12% reduction in epithelial thickness
- at 4-5 months of age, homozygotes display a 40% reduction in stromal thickness

cellular phenotype

increased fibroblast proliferation
- mutant mouse embryonic fibroblasts (MEFs) display increased rates of proliferation relative to wild-type
- in culture, mutant corneal fibroblasts show increased growth relative to wild-type

abnormal apoptosis
- in addition, 6-week-old homozygotes show a significant reduction in apoptosis during stromal wound healing
- at P10, homozygous mutant mice exhibit increased proliferation and decreased apoptosis of stromal keratocytes during postnatal maturation of the cornea

decreased fibroblast apoptosis
- mutant MEFs display reduced apoptosis relative to wild-type

growth/size/body region phenotype

decreased body weight
- homozygotes weigh 70-80% of the body weight of wild-type littermates

decreased body size
- homozygotes are viable and fertile; however, ~10-15% of mutant mice are significantly smaller at birth

References

Chakravarti S; Magnuson T; Lass JH; Jepsen KJ; LaMantia C; Carroll H. 1998. Lumican regulates collagen fibril assembly: skin fragility and corneal opacity in the absence of lumican. J Cell Biol 141(5):1277-86PubMed: 9606218 MGI: J:48068

Additional - Lum^tm1Chak related

Technical Support

CONTACT TECHNICAL SUPPORT

Genotyping Protocols
- Separated PCR:Lum
- Genotyping resources and troubleshooting
Breeding Considerations

When maintaining a live colony, female heterozygous mice can be bred to male homozygous mice. Although homozygotes are viable and fertile, homozygous females are not good breeders.
- Additional Breeding and Husbandry Support
Citation
When using the B6.129S-Lum^tm1Chak/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #019749 in your Materials and Methods section.

Animal Health Reports

Facility Barrier Level Descriptions

Production of mice from cryopreserved embryos or sperm occurs in a maximum barrier room, G200

Pricing & Availability

Cryo Recovery

Domestic
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Live Mouse

Age

Genotype

Price

weeks

Cryorecovery - Pricing

Service/Product	Description	Price
	Heterozygous or wildtype for Lum<tm1Chak>

Related Products and Services

Frozen Mouse Embryo	B6.129S-Lum<tm1Chak>/J Frozen Embryo	$2595.00
Frozen Mouse Embryo	B6.129S-Lum<tm1Chak>/J Frozen Embryo	$2595.00
Frozen Mouse Embryo	B6.129S-Lum<tm1Chak>/J Frozen Embryo	$3373.50
Frozen Mouse Embryo	B6.129S-Lum<tm1Chak>/J Frozen Embryo	$3373.50

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The Jackson Laboratory's Genotype Promise

The Jackson Laboratory has rigorous genetic quality control and mutant gene genotyping programs to ensure the genetic background of JAX® Mice strains as well as the genotypes of strains with identified molecular mutations. JAX® Mice strains are only made available to researchers after meeting our standards. However, the phenotype of each strain may not be fully characterized and/or captured in the strain data sheets. Therefore, we cannot guarantee a strain's phenotype will meet all expectations. To ensure that JAX® Mice will meet the needs of individual research projects or when requesting a strain that is new to your research, we suggest ordering and performing tests on a small number of mice to determine suitability for your particular project. We do not guarantee breeding performance and therefore suggest that investigators order more than one breeding pair to avoid delays in their research.

Terms Of Use

General Terms and Conditions

Questions about Terms of Use

Additional Use Restrictions Apply

Use of MICE by companies or for-profit entities requires a license prior to shipping.

Licensing Information

Phone: 207-288-6470

Email: TechTran@jax.org

Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Control Suggestions

Additional Information

Selected References

Lumtm1Chak

Allele Symbol: Lumtm1Chak

Disease Terms

Model with phenotypic similarity to human disease where etiologies are distinct. Human genes are associated with this disease. Orthologs of these genes do not appear in the mouse genotype(s).

Research Areas By Phenotype

This mouse can be used to support research in many areas including:

Mammalian Phenotype Terms by Genotype

Genotype: Lumtm1Chak/Lumtm1Chakinvolves: 129S/Sv * CD-1

cardiovascular system phenotype

integument phenotype

mammalian phenotype

muscle phenotype

skeleton phenotype

vision/eye phenotype

cellular phenotype

growth/size/body region phenotype

References

Additional - Lumtm1Chak related

Genotyping Protocols

Breeding Considerations

Citation

Animal Health Reports

Production of mice from cryopreserved embryos or sperm occurs in a maximum barrier room, G200

Live Mouse

Cryorecovery - Pricing

Related Products and Services

Payment Terms and Conditions

The Jackson Laboratory's Genotype Promise

Terms Of Use

Additional Use Restrictions Apply

Licensing Information

Lum^tm1Chak

Allele Symbol: Lum^tm1Chak

Genotype: Lum^tm1Chak/Lum^tm1Chak
involves: 129S/Sv * CD-1

Additional - Lum^tm1Chak related