Description

Strain Information

Type Congenic; Mutant Strain; Targeted Mutation; Additional information on Genetically Engineered Mutant Mice. Species laboratory mouse   Donating Investigator Michael Wilson,   University of New Mexico

Description
Homozygous mutant mice die at birth from respiratory failure. At embryonic day 17.5 to 18.5 homozygous embryos appear smaller and do not display spontaneous movement or sensorimotor reflexes. Dilated vascular channels in subcutaneous tissues give the embryos an external blotchy appearance. No gene product (mRNA or protein) is detected by Northern or Western blot analysis of brain tissue from homozygous animals. Histological analysis of fetal diaphragm tissue reveals a dispersed pattern of innervation and fewer layers of muscle fibers. Thin, disarrayed intercostal and anterior chestwall muscles are also observed. Spontaneous miniature endplate potential (mEPP) activity is detected in the diaphragm phrenic nerve, but no evoked endplate potentials (EPP), evoked neurotransmitter release or muscle contraction is detected with stimulation of the neuromuscular junction (NMJ). Mutant NMJs exhibit larger endplate diameters and lower levels of acetylcholinesterase. Tetrodotoxin (TTX) resistant miniature excitatory postsynaptic currents (mEPSCs), but not evoked, action-potential dependent responses are detected from mutant central nervous system (CNS) synapses. Mice that are heterozygous for the targeted mutation are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities. Thismutant mouse strain may be useful in studies of neuroexocytosis and neurotransmitter release in the developing nervous system.

Development
A targeting vector containing neomycin resistance and herpes simplex virus thymidine kinase genes was used to disrupt exons 5a and 5b and part of the downstream intron. The construct was electroporated into 129X1/SvJ derived embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts. The resulting chimeric animals were crossed to C57BL/6 mice, and then backcrossed to C57BL/6 for 7 generations (September 2003).

Control Information

	Control
	Wild-type from the colony
	000664 C57BL/6J
		Wildtype mice from the colony or C57BL/6J mice (Stock No. 000664) may be used as controls.
Considerations for Choosing Controls

Additional Web Information

Genetic Quality Control Annual Report

Phenotype

Phenotype Information

View Mammalian Phenotype Terms

Mammalian Phenotype Terms

      assigned by genotype

Snap25^tm1Mcw/Snap25^tm1Mcw

        involves: C57BL/6

• lethality-prenatal/perinatal
• neonatal lethality (MGI Ref ID J:73755)
  • lethality at birth is suggested to be due to respiratory failure
• behavior/neurological phenotype
• no spontaneous movement (MGI Ref ID J:73755)
  • no movement was detectable in response to mechanical stimuli
• unresponsive to tactile stimuli (MGI Ref ID J:73755)
• cardiovascular system phenotype
• abnormal vasculature (MGI Ref ID J:73755)
  • dilated vascular channels are found in the subcutaneous soft tissue resulting in hyperemic skin blotches
• embryogenesis phenotype
• reduced embryo size (MGI Ref ID J:73755)
  • homozygous animals appear smaller and exhibit a tucked position
• growth/size phenotype
• reduced embryo size (MGI Ref ID J:73755)
  • homozygous animals appear smaller and exhibit a tucked position
• muscle phenotype
• abnormal intercostal muscle morphology (MGI Ref ID J:73755)
  • thin and disorganized musculature and AChR cluster sites are more dispersed in mutant muscles
• thin diaphragm muscle (MGI Ref ID J:73755)
  • thin and disorganized musculature and AChR cluster sites are more dispersed in mutant muscles
• respiratory system phenotype
• respiratory failure (MGI Ref ID J:73755)
  • at birth, animals do not breathe
• skin/coat/nails phenotype
• abnormal skin condition (MGI Ref ID J:73755)
• blotchy skin (MGI Ref ID J:73755)
  • dilated vascular channels are found in the subcutaneous soft tissue resulting in hyperemic skin blotches
• touch/vibrissae phenotype
• unresponsive to tactile stimuli (MGI Ref ID J:73755)
• nervous system phenotype
• abnormal CNS synaptic transmission (MGI Ref ID J:73755)
• abnormal miniature excitatory postsynaptic currents (MGI Ref ID J:73755)
  • spontaneous miniature excitatory postsynaptic currents (mEPSCs) were detected in mutant hippocampal neurons, but the frequency was lower than in controls; treatment of neurons with agents to accelerate vesicle fusion events increased this frequency
  • neuronal postsynaptic responses were unaffected
  • the transmission defect is suggested to be due to a block in regulated presynaptic exocytosis
• abnormal PNS synaptic transmission (MGI Ref ID J:73755)
• abnormal endplate potential (MGI Ref ID J:73755)
  • phrenic nerve stimulation of mutant diaphragms resulted in absence of EPPs and evoked contractility; however, carbachol treatment resulted in contraction of mutant diaphragms suggesting that the muscles are capable of responding to neurotransmitter signals
  • spontaneous miniature EPP activity was recorded in mutant diaphragms due to low spontaneous release of acetylcholine
• abnormal brain morphology (MGI Ref ID J:80415)
• abnormal cerebral cortex morphology (MGI Ref ID J:80415)
  • abnormal neocortical plate morphology; irregular bulges and undulations in the cortical plate in a subset of mutant animals

View Research Applications

Research Applications

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

Snap25^tm1Mcw related

Cardiovascular Research
Vascular Defects

Developmental Biology Research
Growth Defects Growth Defects (homozygous)
Perinatal Lethality (Homozygous)

Neurobiology Research
Neuromuscular Defects

Genes & Alleles

Gene & Allele Information

Allele Symbol		Snap25tm1Mcw
Allele Name		targeted mutation 1, Michael C Wilson
Allele Type		Targeted (knock-out)
Common Name(s)		Snap25-;
Mutation Made By		Michael Wilson,   University of New Mexico
Gene Symbol and Name		Snap25, synaptosomal-associated protein 25
Chromosome		2
Gene Common Name(s)		Bdr; FLJ23079; GENA 70; MGC105414; RIC-4; RIC4; SEC9; SNAP; SNAP-25; SNAP-25B; bA416N4.2; blind drunk; dJ1068F16.2; sp;
Molecular Note		Exon 5a/b and part of the downstream intron were replaced with a PGK-neo cassette by homologous recombination. Western blot of brain proteins and RT-PCR analysis of total brain RNA from homozygous mutant embryos verified the absence of gene expression. [MGI Ref ID J:73755]

Genotyping

Genotyping Information

This strain will not have a genotyping protocol or one is not currently available.

Helpful Links

Optimizing PCR Protocols

References

References

Selected Reference(s)

Washbourne P; Thompson PM; Carta M; Costa ET; Mathews JR; Lopez-Bendito G; Molnar Z; Becher MW; Valenzuela CF; Partridge LD; Wilson MC. 2002. Genetic ablation of the t-SNARE SNAP-25 distinguishes mechanisms of neuroexocytosis. Nat Neurosci 5(1):19-26. [PubMed: 11753414]  [MGI Ref ID J:73755]

Additional References

Molnar Z; Lopez-Bendito G; Small J; Partridge LD; Blakemore C; Wilson MC. 2002. Normal development of embryonic thalamocortical connectivity in the absence of evoked synaptic activity. J Neurosci 22(23):10313-23. [PubMed: 12451131]  [MGI Ref ID J:80415]

Snap25^tm1Mcw related

Bark C; Bellinger FP; Kaushal A; Mathews JR; Partridge LD; Wilson MC. 2004. Developmentally regulated switch in alternatively spliced SNAP-25 isoforms alters facilitation of synaptic transmission. J Neurosci 24(40):8796-805. [PubMed: 15470145]  [MGI Ref ID J:94005]

Molnar Z; Higashi S; Lopez-Bendito G. 2003. Choreography of early thalamocortical development. Cereb Cortex 13(6):661-9. [PubMed: 12764042]  [MGI Ref ID J:102089]

Molnar Z; Lopez-Bendito G; Small J; Partridge LD; Blakemore C; Wilson MC. 2002. Normal development of embryonic thalamocortical connectivity in the absence of evoked synaptic activity. J Neurosci 22(23):10313-23. [PubMed: 12451131]  [MGI Ref ID J:80415]

Nagy G; Milosevic I; Fasshauer D; Muller EM; de Groot BL; Lang T; Wilson MC; Sorensen JB. 2005. Alternative splicing of SNAP-25 regulates secretion through nonconservative substitutions in the SNARE domain. Mol Biol Cell 16(12):5675-85. [PubMed: 16195346]  [MGI Ref ID J:107104]

Pozzi D; Condliffe S; Bozzi Y; Chikhladze M; Grumelli C; Proux-Gillardeaux V; Takahashi M; Franceschetti S; Verderio C; Matteoli M. 2008. Activity-dependent phosphorylation of Ser187 is required for SNAP-25-negative modulation of neuronal voltage-gated calcium channels. Proc Natl Acad Sci U S A 105(1):323-8. [PubMed: 18162553]  [MGI Ref ID J:131023]

Tafoya LC; Mameli M; Miyashita T; Guzowski JF; Valenzuela CF; Wilson MC. 2006. Expression and function of SNAP-25 as a universal SNARE component in GABAergic neurons. J Neurosci 26(30):7826-38. [PubMed: 16870728]  [MGI Ref ID J:111062]

Health & husbandry

Health & Colony Maintenance Information

Colony Maintenance

Breeding & Husbandry	This strain originated on a B6;129 background and was backcrossed to C57BL/6 for 7 generations (September 2003). The strain must be maintained as a heterozygote. Homozygotes are not viable.

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
	Wild-type from the colony
	000664 C57BL/6J
		Wildtype mice from the colony or C57BL/6J mice (Stock No. 000664) may be used as controls.
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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Contact Information
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Tel: 800.422.6423 or 207.288.5845
Fax: 207.288.6150
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Terms of Use

Terms of Use

General Terms and Conditions

For Licensing and Use Restrictions view the link(s) below:
- Use of MICE by companies or for-profit entities requires a license prior to shipping.

Contact information

General inquiries

Contracts Administration

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

JAX^® Mice & Services Conditions of Use

“Each recipient institution, including its employees and other researchers under its control (RECIPIENT), of mice or services using mice from The Jackson Laboratory (TJL) agrees that such mice, descendants of those mice derived by inbreeding or crossbreeding, including unmodified derivatives of those mice or their descendants (“MICE”) shall not be: (i) used for any purpose other than the internal research of the RECIPIENT, (ii) sold or otherwise provided to any third party for any use, or (iii) provided to any agent or other third party to provide breeding or other services with respect to MICE. Acceptance of MICE from TJL shall be deemed agreement by RECIPIENT to these conditions, and departure from these conditions requires The Jackson Laboratory’s prior written authorization.”

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MICE and biological materials are to be used in a safe manner and in accordance with all applicable governmental rules and regulations.

The foregoing represents the General Terms and Conditions applicable to The Jackson Laboratory’s MICE, products and services. In addition, special terms and conditions of sale of certain MICE, products and services may be set forth separately in The Jackson Laboratory web pages, catalogs, price lists, contracts, and/or other documents, and these special terms and conditions shall also govern the sale of these MICE, products and services by The Jackson Laboratory, and by its licensees and distributors.

Acceptance of delivery of MICE, products or services shall be deemed agreement to these terms and conditions. No purchase order or other document transmitted by purchaser or recipient that may modify the terms and conditions hereof, shall be in any way binding on The Jackson Laboratory, and instead the terms and conditions set forth herein, including any special terms and conditions set forth separately, shall govern the sale of MICE, products services by The Jackson Laboratory.