Overview

These TRPM8^EGFPf targeted mutation (knock in) mice display deficits in cool thermosensation and may be useful in studies of cold nociception.

Donating Investigator

Ardem Pataoputian, The Scripps Research Institute

Genetic overview

Genetic Background	Generation

Trpm8^tm1Apat

Allele Type	Gene Symbol	Gene Name
Targeted (Reporter, Null/Knockout)	Trpm8	transient receptor potential cation channel, subfamily M, member 8

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Research Applications

Neurobiology Research
Research Tools
Sensorineural Research

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Base Price

Starting at:

$2,595.00 Domestic price Cryo Recovery

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Details

Detailed Description

These mutant mice express farnesylated enhanced GFP gene (EGFPf) from the endogenous Trpm8 locus. Mice that are homozygous for the targeted mutation are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities. No wildtype gene product (mRNA) is detected although low levels of a truncated mRNA splice variant is sometimes detected by RT-PCR analysis of total RNA derived from dorsal root ganglia (DRG) isolated from homozygotes. No Trpm8 locus mRNA is detected by in situ hybridization of DRG isolated from homozygotes. In thermotaxis assays performed in a cage equipped with surface temperature gradients, homozygous mice spend less time in the warmer area compared to wildtype controls. Homozygotes display diminished responses to application of acetone and injection of icilin. Noxious cold sensation of colder temperatures (sub zero) is normal. Mutant mice do not display cold-induced analgesia in response to pain. EGFPf-positive neurons isolated from homozygotes are more sensitive to cold and menthol. TRPM8-expressing neurons isolated from mutants are specifically labeled by EGFPf expression. EGFPf fluorescence is detected in the dorsal root ganglia and spinal cord from both homozygotes and heterozygotes (with an increased fluorescence intensity observed in homozygotes). Axonal terminals are visualized utilizing immunohistochemical methods.

Development

A targeting vector containing a farnesylated enhanced GFP gene (EGFP-F), SV40polyA signal sequence, and a loxP site flanked EM7-PGK-Neo cassette, was used to replace sequence encoding amino acids 2-29. The construct was electroporated into 129S1/Sv-Oca2⁺ Tyr⁺ Kitl⁺ derived CJ7 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts. The resulting male chimeric animals were crossed to C57BL/6 female mice. The mice were crossed to an EIIA-Cre deleter strain (Stock No. 003724) to remove the floxed Neo cassette. Upon arrival at The Jackson Laboratory, the mice were crossed to C57BL/6J (Stock No. 000664) at least once to establish the colony.

Expression Data

Expressed Gene	GFP, Green Fluorescent Protein,
Site of Expression	Farnesylated EGFP (EGFPf) fluorescence is detected in the dorsal root ganglia and spinal cord.

Control Suggestions

101043 B6129SF1/J, ()
101045 B6129SF2/J, ()

Additional Information

Considerations for Choosing Controls

Selected References

Dhaka A; Murray AN; Mathur J; Earley TJ; Petrus MJ; Patapoutian A. 2007. TRPM8 is required for cold sensation in mice. Neuron 54(3):371-8PubMed: 17481391 MGI: J:122963

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Genetics

Trpm8^tm1Apat

Allele Symbol: Trpm8^tm1Apat

Allele Name	targeted mutation 1, Ardem Patapoutian
Allele Type	Targeted (Reporter, Null/Knockout)
Allele Synonym(s)	Trpm8^tm1Apat; targeted mutation 1, Ardem Patapoutian
Gene Symbol and Name	Trpm8, transient receptor potential cation channel, subfamily M, member 8
Gene Synonym(s)	CMR1; TRPP8; Trp-p8; LTRPC6; trp-p8; LTrpC-6
Expressed Gene	GFP, Green Fluorescent Protein,
Site of Expression	Farnesylated EGFP (EGFPf) fluorescence is detected in the dorsal root ganglia and spinal cord.
Strain of Origin	129S1/Sv-Oca2⁺ Tyr⁺ Kitl⁺
Chromosome	1
Molecular Note	Velocigene technology was used to delete sequences encoding amino acid residues 2-29 and insert a farnesylated enhanced GFP gene followed by an SV40polyA tail in frame with the start codon of the endogenous gene. RT-PCR confirmed absence of wild-type transcript in mutants, however, in some samples a truncated splice variant was detected at low levels.
Mutations Made By	Ardem Patapoutian, The Scripps Research Institute

Disease/Phenotype

Disease Terms

Research Areas By Genotype

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

Neurobiology Research
- Fluorescent protein expression in neural tissue
Research Tools
- Fluorescent Proteins
Sensorineural Research
- Nociception

Mammalian Phenotype Terms by Genotype

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

Genotype: Trpm8^tm1Apat/Trpm8^tm1Apat
involves: 129S1/Sv * C57BL/6

behavior/neurological phenotype

abnormal nociception after inflammation
- absence of cold-induced analgesic response following formalin injection

abnormal thermosensation
- mice show no preference for a floor at 31 C over one at 18 C unlike wild-type mice
- presented with a floor temperature gradient mice spend more time in the cooler areas (23 - 30 C) and less time in warm areas (30 - 38 C) compared to wild-type mice
- response of mice to the cooling effect of acetone is significantly reduced

increased thermal nociceptive threshold
- following icilin injection mice do not display hindpaw withdrawal on a 1 C plate or wet-dog shakes
- Normal - however, cold nociception response to sub-zero temperatures is normal

nervous system phenotype

abnormal neuron physiology
- only 7.6% of cultured dorsal root ganglion neurons respond to a cold stimulus compared to 14.9% of wild-type neurons

integument phenotype

abnormal nociception after inflammation
- absence of cold-induced analgesic response following formalin injection

increased thermal nociceptive threshold
- following icilin injection mice do not display hindpaw withdrawal on a 1 C plate or wet-dog shakes
- Normal - however, cold nociception response to sub-zero temperatures is normal

References

Dhaka A; Murray AN; Mathur J; Earley TJ; Petrus MJ; Patapoutian A. 2007. TRPM8 is required for cold sensation in mice. Neuron 54(3):371-8PubMed: 17481391 MGI: J:122963

Additional - Trpm8^tm1Apat related

Technical Support

Contact Technical Support

Genotyping Protocols
- Probe: Gt(rosa)26sor^Tm1Sor Probe
- Separated MCA: Gt(ROSA)26Sor^tm1sor STD
- Standard PCR: Trpm8^tm1Apat
- Genotyping resources and troubleshooting
Breeding Considerations

When maintaining a live colony, these mice can be bred as homozygotes.
- Additional Breeding and Husbandry Support
Citation
When using the STOCK Trpm8^tm1Apat/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #014581 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	Genotype	Price
	Heterozygous or Wild-type for Trpm8<tm1Apat>

We will fulfill your order by providing at least two carriers for each strain ordered. The total number, sex, and genotypes provided will vary, although typically 8 or more animals are provided. Please check genotypes which will be recovered. While the genotypes of all animals produced will be communicated to you prior to scheduling shipment, the genotypes of animals provided may not reflect the mating scheme and genotypes described in the strain description. Animals are typically ready to ship in 11-14 weeks. If a second recovery is required to produce the minimum number of animals, then delivery time would increase to approximately 25 weeks. If we fail to produce animals of the correct genotype, you will not be charged. We cannot guarantee the reproductive success of mice shipped to your facility. If the mice are lost after the first three days (post-arrival) or do not produce progeny at your facility, a new order and fee will be necessary.

Cryorecovery to establish a Dedicated Supply for greater quantities of mice. Mice recovered can be used to establish a dedicated colony to contractually supply you mice according to your requirements. Price by quotation.

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Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Expression Data

Control Suggestions

Additional Information

Selected References

Trpm8tm1Apat

Allele Symbol: Trpm8tm1Apat

Disease Terms

Research Areas By Genotype

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

Mammalian Phenotype Terms by Genotype

Genotype: Trpm8tm1Apat/Trpm8tm1Apatinvolves: 129S1/Sv * C57BL/6

behavior/neurological phenotype

nervous system phenotype

integument phenotype

References

Additional - Trpm8tm1Apat 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

JAX® Mice, Products & Services Conditions of Use

No Warranty

Credit for PRODUCTS or SERVICES

Animal Care and Use for SERVICES

No Liability

Trpm8^tm1Apat

Allele Symbol: Trpm8^tm1Apat

Genotype: Trpm8^tm1Apat/Trpm8^tm1Apat
involves: 129S1/Sv * C57BL/6

Additional - Trpm8^tm1Apat related