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Strain Detail Sheet

Strain Name:
B6.129-Htr2ctm1Knk/Mmmh
Stock Number:
031761-MU
Citation ID:
RRID:MMRRC_031761-MU

Strain Information

Gene Details

Htr2ctm1Knk
Name: 5-hydroxytryptamine (serotonin) receptor 2C; targeted mutation 1, Kazuko Nishikura
Synonyms: 5-HT2CR-VGV
Type: Allele
Species: Mus musculus (mouse)
Chromosome:
Alteration at locus: Knock-In
Htr2c
Name: 5-hydroxytryptamine (serotonin) receptor 2C
Synonyms: 5-HT2C receptor, 5-HT2cR, Htr1c, SR1, 5HT1c
Type: Gene
Species: Mus musculus (mouse)
Chromosome: X
Alteration at locus: Knock-In
NCBI: 15560
HGNC: HGNC:5295
Homologene: 20242

Additional Resources
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Genetic Alterations
Exon 5 was replaced with one in which 5 adenosines were converted to guanosines by nucleotide substitutions giving rise to I156V, N158G and I160V amino acid substitutions. A floxed neo cassette was inserted upstream of exon 5 for selection purposes. Mice only express the isoform carrying these substitutions.

Genotype Determination
ES Cell Line
R1

Strain Description

Phenotype
Homozygous: Although INI mice grew normally, VGV mice had a severely reduced fat mass, in spite of compensatory hyperphagia, due to constitutive activation of the sympathetic nervous system and increased energy expenditure. Furthermore, serotonergic neurotransmission was oversensitized in VGV mice, most likely due to the increased cell surface expression of VGV receptors. In addition, both VGV and INI mice display behavioral changes (more nervousness).

More specifically, RNA editing of the serotonin 2C receptor (5-HT2CR) gene replaces gene-encoded Ile, Asn, Ile (INI) with Val, Gly, Val (VGV) by converting adenosine to inosine, producing up to 24 isoforms across brain regions, with knock-in mouse lines expressing either fully edited 5-HT2CR-VGV or unedited 5-HT2CR-INI to investigate its physiological role. The 5-HT2CR-VGV knock-in vector introduced five A-G mutations at editing sites A-E, while the 5-HT2CR-INI vector deleted the 160 bp editing complementary sequence in intron 5, with a silent XbaI site in exon 5 enabling mutated allele screening in ES cells and mice. Inosine, read as guanosine during translation, drives these changes, and ES cell clones were selected for G418 resistance and ganciclovir sensitivity, screened via PCR.


Hemizygous: Heterozygous female VGV mice has about half reduced body weight of homozygous female mice.
Mammalian Phenotype Terms
Allelic Composition: Htr2ctm1Knk/Htr2ctm1Knk (Genetic Background: B6.Cg-Htr2ctm1Knk )
Allelic Composition: Htr2ctm1Knk/Htr2ctm1Knk (Genetic Background: C.Cg-Htr2ctm1Knk )
Allelic Composition: Htr2ctm1Knk/Htr2c+ (Genetic Background: B6.Cg-Htr2ctm1Knk )
Allelic Composition: Htr2ctm1Knk/Htr2ctm2Knk (Genetic Background: B6.Cg-Htr2ctm1Knk/Htr2ctm2Knk )
MeSH Terms
  • Adipose Tissue/metabolism
  • Amino Acid Sequence/genetics
  • Animals
  • Energy Metabolism/genetics
  • Female
  • Gene Knock-In Techniques
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Mutation/genetics
  • Protein Isoforms/genetics
  • RNA Editing/genetics
  • RNA, Messenger/genetics
  • Receptor, Melanocortin, Type 4/genetics
  • Receptor, Melanocortin, Type 4/metabolism
  • Receptor, Serotonin, 5-HT2C/genetics
  • Serotonin/metabolism
  • Signal Transduction/genetics
  • Sympathetic Nervous System/physiopathology
  • Synaptic Transmission/genetics
Strain Development
Inosine, interpreted as guanosine by the translational machinery, drives RNA editing in the 5-HT2CR gene, where the VGV knock-in vector incorporates five A→G mutations at sites A-E, while the INI vector deletes the 160 bp editing complementary sequence in intron 5, with a silent XbaI site in exon 5 aiding mutated allele screening in ES cells and knock-in mice. ES cell clones, selected for G418 resistance and ganciclovir sensitivity, were screened by Southern hybridization with 5′ and 3′ 5-HT2CR and Neo gene probes or by PCR using intron 4 (5′-ACTGATACTACCATCACTGG-3′) and intron 5 (5′-AGCATATATAGGAAATTGCAGTAACCCT-3′) primers followed by XbaI digestion, identifying three positive clones. One clone generated germline-transmitting male chimeric founders, establishing VGV and INI alleles in hemizygous male and heterozygous/homozygous female mice on C57BL/6J.
Founder male mice were backcrossed to wt C57BL/6 female mice. More than 10 backcrossing were done.


Suggested Control Mice

Wildtype littermates

MMRRC Genetic QC

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 mmrrc@missouri.edu. Older strains may not have this information.

Research Applications

  • Diabetes/Obesity
  • Metabolism
  • Models for Human Disease
  • Neurobiology

Strain Origin

Donor
Kazuko Nishikura, Ph.D., The Wistar Institute
Primary Reference
Kawahara Y, Grimberg A, Teegarden S, Mombereau C, Liu S, Bale TL, Blendy JA, Nishikura K. Dysregulated editing of serotonin 2C receptor mRNAs results in energy dissipation and loss of fat mass. J Neurosci. 2008 Nov 26;28(48):12834-44. (Medline PMID: 19036977)

Colony and Husbandry Information

Special Considerations

When hemizygous VGV male are mated with wt or heterozygous female mice, because of small body size of hemizygous male and homozygous female, wt or heterozygous female mice need to remove from the colony within 2-3 days of birth. Otherwise, hemizygous male and homozygous female pups often die within the first week of birth (social competition).

Health Status Report

Colony Surveillance Program and Current Health 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 mmrrc@missouri.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.

Cryopreserved material may be available upon request, please inquire to mmrrc@missouri.edu for more information.

Conditions of Distribution

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 $)
*Shipping & Handling not included*		 Units Notes
031761-MU-SPERM Cryo-preserved spermatozoa $437.00 / Non-Profit Aliquot Approximate quantity3
031761-MU-RESUS Litter recovered from cryo-archive $2,624.00 / Non-Profit Litter Recovered litter4; additional fees for any special requests.
Cryopreserved material may be available upon request, please inquire to mmrrc@missouri.edu 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.