Description

Strain Information

Type Mutant Stock; Targeted Mutation; Additional information on Genetically Engineered Mutant Mice. Species laboratory mouse Generation F?+N1F1+N1p (07-AUG-05)   Donating Investigator Peter Mombaerts,   Max Planck Institute of Biophysics

Description
These mutant mice express synapto-pHluorin (spH) from the endogenous locus encoding the olfactory marker protein (OMP) as a result of a targeted knockin mutation that replaces the OMP coding region with that of spH. The OMP locus directs expression of spH at high levels and exclusively in mature sensory neurons of the main olfactory epithelium (OSNs) and the vomeronasal epithelium. SpH is a pH-sensitive variant of the green fluorescent protein (ecliptic pHluorin) fused to the mouse synaptic vesicle-associated protein (VAMP2). The protein is localized preferentially in synaptic vesicles, but is also present on the plasma membrane of OSN axons and nerve terminals. The fluorescent domain of spH is exposed to the acidic lumen of synaptic vesicles where it is ~ 20 fold less fluorescent than at neutral pH. Upon synaptic release, the lumen of the synaptic vesicles becomes continuous with the extracellular space resulting in an increase in pH that causes a rapid increase in fluorescence. In mutant mice all glomeruli of the olfactory bulb are robustly labeled. There are ~1800 glomeruli in the main olfactory bulb, and each glomerulus receives axonal input from thousands of OSNs that express the same odorant receptor. Glomeruli in the dorsal surface of the olfactory bulb can be observed in vivo, after thinning the skull, through a standard epifluorescence microscope. Fluorescence changes caused by neuronal activity and presynaptic release can be measured in living mice following odorant stimulation. Mice that are homozygous for this allele do not express gene product (OMP) but are viable, fertile, normal in size and do not display gross physical or behavioral abnormalities. Homozygous mice produce small litters and breed at reduced frequency. This mutant mouse strain should prove useful to measure stimulus-induced activity or presynaptic vesicle release in OSNs of living mice.

Development
A targeting vector containing super-ecliptic synapto-pHluorin coding sequence and an auto-excising neomycin resistance cassette was used to replace the Omp coding region sequence in 129P2/OlaHsd-derived E14 embryonic stem (ES) cells. The loxP-flanked auto-excising neomycin resistance cassette is composed of the mouse angiotensin-converting enzyme promoter (tACE) located 5' of sequences encoding Cre recombinase and the neomycin resistance gene. The angiotensin-converting enzyme promoter has been shown to initiate transcription only during spermatogenesis. Correctly targeted ES cells were injected into C57BL/6 blastocysts and a chimeric animal derived from cell clone OSE-80 was obtained.

The plasmid that was used to generate this mutation is banked and described at Addgene.

Control Information

	Control
	100903 B6129PF2/J	(approximate)
Considerations for Choosing Controls

Related Strains

Strains carrying other alleles of Omp

007768	B6.129S2-Omptm1Mom/MomJ
006667	B6;129P2-Omptm3Mom/MomJ
006668	B6;129P2-Omptm4(cre)Mom/MomJ
006594	B6;129S2-Omptm1Mom/MomJ

View Strains carrying other alleles of Omp     (4 strains)

Phenotype

Phenotype Information

View Mammalian Phenotype Terms

Mammalian Phenotype Terms

      assigned by genotype

Omp^tm2(spH)Mom/?

        involves: 129P2/OlaHsd * C57BL/6

• no phenotypic analysis
• *normal* no phenotypic analysis (MGI Ref ID J:88765)
  • presence of this allele was used to investigate odorant-evoked synaptic transmissions; no phenotypic analysis was done

View Research Applications

Research Applications

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

Neurobiology Research
Fluorescent protein expression in neural tissue

Research Tools
Genetics Research (Tissue/Cell Markers: neurons)
Neurobiology Research (cell marker)
Sensorineural Research

Genes & Alleles

Gene & Allele Information

Allele Symbol		Omptm2(spH)Mom
Allele Name		targeted mutation 2, Peter Mombaerts
Allele Type		Targeted (Reporter)
Common Name(s)		OMP-spH; OMP-synapto-pHluorin;
Mutation Made By		Peter Mombaerts,   Max Planck Institute of Biophysics
Strain of Origin		129P2/OlaHsd
ES Cell Line Name		E14
ES Cell Line Strain		129P2/OlaHsd
Site of Expression		active mature sensory neurons of the main olfactory epithelium (OSNs) and the vomeronasal epithelium; protein localized preferentially in synaptic vesicles, but is also present on the plasma membrane of OSN axons and nerve terminals; all glomeruli of the olfactory bulb are robustly labeled (visible in vivo); pH sensitive
Gene Symbol and Name		Omp, olfactory marker protein
Chromosome		7
General Note		In the acidic lumen of pre-synaptic vesicles, the fluorescence of spH is only about 5% of its intensity at neutral pH. When synaptic vesicles with spH anchored to their lumenal face fuse with the plasma membrane to release their contents, the spH is exposed to the pH-neutral extracellular environmment and rapidly becomes highly fluorescent. All olfactory bulb glomeruli fluoresce, with varying intensity. The superficial nerve layer overlying the glomerular layer and comprising axons of olfactory sensory neurons is more weakly fluorescent. In vivo fluorescent imaging of the olfactory bulb demonstrates that odorant-evoked responses cause strong fluorescence of odorant-specific, bilaterally symmetric groups of glomeruli.
Molecular Note		The coding exon was replaced by a construct containing the ecliptic synaptopHluorin (spH) coding sequence and a self-excising neomycin resistance cassette. SpH is a fusion protein joining a pH-sensitive derivative of wild-type green fluorescent protein (GFP) to the carboxyl terminus of a mouse synaptic vesicle-associated membrane protein (VAMP2), which targets spH to the lumenal face of pre-synaptic vesicles. The loxP-flanked, male-germline self-excising neomycin resistance cassette (which was deleted inthe chimeric founder's germline) contains the Cre recombinase coding sequence under control of the mouse testis-specific angiotensin-converting enzyme (tACE) promoter and tk-neo. The endogenous gene regulatory sequences direct spH/Vamp expression specifically to sensory neurons in glomeruli of the olfactory bulb. [MGI Ref ID J:36595] [MGI Ref ID J:79620] [MGI Ref ID J:88765]

Genotyping

Genotyping Information

Genotyping Protocols

Omp^tm2(spH)Mom, STD PCR, vers. 1

Helpful Links

Optimizing PCR Protocols

References

References

Selected Reference(s)

Bozza T; McGann JP; Mombaerts P; Wachowiak M. 2004. In vivo imaging of neuronal activity by targeted expression of a genetically encoded probe in the mouse. Neuron 42(1):9-21. [PubMed: 15066261]  [MGI Ref ID J:88765]

Additional References

Omp^tm2(spH)Mom related

Bunting M; Bernstein KE; Greer JM; Capecchi MR; Thomas KR. 1999. Targeting genes for self-excision in the germ line. Genes Dev 13(12):1524-8. [PubMed: 10385621]  [MGI Ref ID J:79620]

McGann JP; Pirez N; Gainey MA; Muratore C; Elias AS; Wachowiak M. 2005. Odorant representations are modulated by intra- but not interglomerular presynaptic inhibition of olfactory sensory neurons. Neuron 48(6):1039-53. [PubMed: 16364906]  [MGI Ref ID J:107569]

Mombaerts P; Wang F; Dulac C; Chao SK; Nemes A; Mendelsohn M; Edmondson J; Axel R. 1996. Visualizing an olfactory sensory map. Cell 87(4):675-86. [PubMed: 8929536]  [MGI Ref ID J:36595]

Wachowiak M; McGann JP; Heyward PM; Shao Z; Puche AC; Shipley MT. 2005. Inhibition [corrected] of olfactory receptor neuron input to olfactory bulb glomeruli mediated by suppression of presynaptic calcium influx. J Neurophysiol 94(4):2700-12. [PubMed: 15917320]  [MGI Ref ID J:127983]

Health & husbandry

Health & Colony Maintenance Information

Colony Maintenance

Breeding & Husbandry	This strain originated on a B6;129P2 background. It is maintained as a homozygote on the same background.
Diet Information	LabDiet® 5K52/5K67

Purchasing information

Pricing, Supply Level & Notes, Controls, General Terms & Conditions

Pricing

Supply Details

Standard Supply

Repository-Cryopreserved. Must Be Recovered. Please refer to pricing and supply notes for further information.

Supply Notes

Cryopreserved Embryos This strain is also available as cryopreserved embryos from our Repository. Orders for cryopreserved embryos are supplied subject to a signed agreement that must be returned to the Customer Service Department after order placement. Experienced technicians at The Jackson Laboratory have recovered frozen embryos of this strain successfully. We will provide you enough embryos to perform two embryo transfers. The Jackson Laboratory does not guarantee successful recovery at your facility. For complete information on purchasing embryos from our repository, please visit our Cryopreserved Embryos web page. Cryorecovery - Standard. The recovery process begins when a signed agreement form is returned to the Customer Service Department after order placement. Although results vary by strain, at least two males and two females (two pairs) will be provided, typically within 15 weeks of our receipt of the signed agreement form. If the first recovery attempt is unsuccessful or only one pair is recovered, a second recovery will be done, extending the delivery time to approximately 25 weeks. At least one member of each pair will be of known genotype and will carry the mutation if it is a mutant strain. Please note that pairs may not reflect the mating scheme utilized by The Jackson Laboratory prior to cryopreservation of the strain. Mating schemes are sometimes modified for successful cryopreservation. Price represents a repository maintenance fee, which includes the cost of recovery of the strain from the cryopreservation resource and the periodic replacement of the frozen embryos used for recovery. Cryorecovery to establish a Dedicated Supply for greater quantities of mice. One to two pairs will be recovered to establish a Dedicated Supply of mice. Price by quotation. For more information on Dedicated Supply, please contact JAX® Services, Tel: 1-800-422-6423 or 1-207-288-5845. This strain is included in the Induced Mutant Resource Colony collection. Genomic DNA is available for this strain from the Mouse DNA Resource.

Control Information

	Control
	100903 B6129PF2/J	(approximate)
Considerations for Choosing Controls
USA, Canada and Mexico - Control Pricing Information for Genetically Engineered Mutant Strains.
International - Control Pricing Information for Genetically Engineered Mutant Strains.

General Terms and Conditions

See Terms of Use

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.

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Ordering and Purchasing Information

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Terms of Use

Terms of Use

General Terms and Conditions

Effective September 26, 2007: License Requirements for Strains using Cre-lox Technology only apply in Canada, see Licenses for Strains using Cre-lox Technology.

Contact information

General inquiries

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phone:	207-288-6470
fax:	207-288-6655

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