Overview

These Pak4 knock out mice may be useful for studying the role of Pak4 in embryonic and neuronal development.

Donating Investigator

Audrey Minden, Rutgers University

Genetic overview

Genetic Background	Generation

Pak4^tm1Amin

Allele Type	Gene Symbol	Gene Name
Targeted (Null/Knockout)	Pak4	p21 (RAC1) activated kinase 4

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Research Applications

Developmental Biology Research
Neurobiology Research
Cell Biology Research
Cardiovascular Research

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Base Price

Starting at:

$2,854.50 Domestic price Cryo Recovery

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Details

Detailed Description

In this strain, a neo cassette replaces exon 1 of the p21 protein (Cdc42/Rac)-activated kinase 4 (Pak4) gene, abolishing gene expression. Heterozygotes are viable, fertile, normal in size, and do not display any gross physical abnormalities. Homozygotes die by E11.5 due to fetal heart defects. Pak4 is a member of the group B family of PAK serine/threonine kinases and is expressed early in development in a variety of tissues. It is involved in the formation of filopodia in response to Cdc42, promoting neuronal growth. Improper folding of the caudal neural tubes of homozygous embryos results in the formation of two neural lumens. They also exhibit abnormalities in the development, migration and differentiation of neurons. Specifically, axonal outgrowth was impaired, and neurons failed to migrate to their proper locations. These mice may be useful for studying the role of Pak4 in embryonic and neuronal development.

Development

A targeting vector was designed to replace exon 1 encoding the p21 protein (Cdc42/Rac)-activated kinase 4 (Pak4) gene with a neomycin resistance (neo) cassette. The construct was electroporated into 129S2/SvPas-derived D3 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6J blastocysts and the resulting chimeric males were bred to C57BL/6J females. The donating investigator reported that these mice were then backcrossed to C57BL/6J mice for at least 9 generations to maintain the colony (see SNP note below). Upon arrival at The Jackson Laboratory, mice were bred to C57BL/6J (Stock No. 000664) for at least one generation 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, 3 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

Approximate Controls

000664 C57BL/6J

Additional Information

Considerations for Choosing Controls

Selected References

Qu J; Li X; Novitch BG; Zheng Y; Kohn M; Xie JM; Kozinn S; Bronson R; Beg AA; Minden A. 2003. PAK4 kinase is essential for embryonic viability and for proper neuronal development. Mol Cell Biol 23(20):7122-33PubMed: 14517283 MGI: J:85954

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Genetics

Pak4^tm1Amin

Allele Symbol: Pak4^tm1Amin

Allele Name	targeted mutation 1, Audrey Minden
Allele Type	Targeted (Null/Knockout)
Allele Synonym(s)	PAK4^-
Gene Symbol and Name	Pak4, p21 (RAC1) activated kinase 4
Gene Synonym(s)
Strain of Origin	129
Chromosome	7
General Note	ES cell line = D3 or E14.
Molecular Note	Exon 1, containing the initiating methionine and the critical GTPase-binding domaain, was replaced with a PGK-neo cassette inserted by homologous recombination. Neither transcript nor protein was detected by Northern and Western blot analysis of homozygous mutant brain tissue.
Mutations Made By	Audrey Minden, Rutgers University

Disease/Phenotype

Disease Terms

Research Areas By Phenotype

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

Developmental Biology Research
- Perinatal Lethality
- Embryonic Lethality (Homozygous)
Neurobiology Research
- Ataxia (Movement) Defects
- Behavioral and Learning Defects
- Neural Tube Defects
Cell Biology Research
- Signal Transduction
Cardiovascular Research
- Heart Abnormalities

Mammalian Phenotype Terms by Genotype

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

Genotype: Pak4^tm1Amin/Pak4^tm1Amin
either: (involves: 129P2/OlaHsd * C57BL/6) or (involves: 129S2/SvPas * C57BL/6)

embryo phenotype

decreased embryo size
- at E9.5, mutant embryos are generally smaller than wild-type embryos

abnormal neural tube morphology
- at E9.5, an improper folding appears to occur between the ventral and dorsal parts of the caudal neural tube, resulting in the formation of two neural lumens
- however, the dorsal-ventral patterning appears normal in mutant neural tubes

decreased embryonic neuroepithelium thickness
- at E9.5, the defects in neuroepithelium are not associated with a severe defect in proliferation or an increase in apoptosis, although sporadic TUNEL-positive staining is observed in tissues outside the neuroepithelium at this stage
- at E9.5, the neuroepithelium around the hindbrain and forebrain is strikingly thinner than normal

growth/size/body region phenotype

decreased embryo size
- at E9.5, mutant embryos are generally smaller than wild-type embryos

mortality/aging

embryonic lethality during organogenesis, complete penetrance
- only partly resorbed embryos are detected at E11.5
- no homozygotes are obtained after E10.5, putatively due to a defect in the fetal heart

muscle phenotype

thin myocardium
- at E10.5, homozygotes display thinning of the myocardial walls of the bulbus cordis and ventricles

nervous system phenotype

abnormal brain morphology
- at E10.5, mutant embryos display a large empty area in the head region in H&E-stained sagittal sections
- at E9.5, mutant brains appear wavy, probably due to postmortem collapse of the thin neuroepithelial walls

abnormal nervous system development
- at E9.5, mutant embryos are translucent around the head and neural tube regions

abnormal neural tube morphology
- however, the dorsal-ventral patterning appears normal in mutant neural tubes
- at E9.5, an improper folding appears to occur between the ventral and dorsal parts of the caudal neural tube, resulting in the formation of two neural lumens

abnormal neuronal migration
- at E9.5, mutant spinal cord motor neurons are not located in their proper lateral positions, indicating a corresponding defect in motor neuron migration
- unlike wild-type embryos where neurofilament positive cells are located at the lateral edge of the hindbrain, mutant embryos display a small number of neurofilament positive cells both medially and laterally, suggesting a defect in neuronal migration

decreased embryonic neuroepithelium thickness
- at E9.5, the defects in neuroepithelium are not associated with a severe defect in proliferation or an increase in apoptosis, although sporadic TUNEL-positive staining is observed in tissues outside the neuroepithelium at this stage
- at E9.5, the neuroepithelium around the hindbrain and forebrain is strikingly thinner than normal

abnormal neuron differentiation
- at E9.5, homozygotes exhibit lack of neuronal differentiation and a concomittant reduction in cell size due to impaired axonal outgrowth

abnormal spinal cord interneuron morphology
- at E9.5, homozygotes display failure of developing ventral interneurons to differentiate

abnormal motor neuron morphology
- at E9.5, homozygotes show failure of spinal cord motor neurons to differentiate

abnormal axon extension
- at E9.5, homozygotes display defective axonal outgrowth in the hindbrain, as shown by staining with anti-neurofilament antibody

cardiovascular system phenotype

thin ventricular wall
- at E10.5, homozygotes display thinning of the primitive ventricular wall

thin myocardium
- at E10.5, homozygotes display thinning of the myocardial walls of the bulbus cordis and ventricles

dilated heart
- at E10.5, the atrium and/or sinus venosus region of the heart appears distended and dilated

cellular phenotype

abnormal cell adhesion
- when grown in serum-free medium, cells isolated from E9.5 mutant, but not wild-type, embryos show significant levels of focal adhesions, as evidenced by large clusters of vinculin

abnormal neuron differentiation
- at E9.5, homozygotes exhibit lack of neuronal differentiation and a concomittant reduction in cell size due to impaired axonal outgrowth

abnormal neuronal migration
- unlike wild-type embryos where neurofilament positive cells are located at the lateral edge of the hindbrain, mutant embryos display a small number of neurofilament positive cells both medially and laterally, suggesting a defect in neuronal migration
- at E9.5, mutant spinal cord motor neurons are not located in their proper lateral positions, indicating a corresponding defect in motor neuron migration

abnormal axon extension
- at E9.5, homozygotes display defective axonal outgrowth in the hindbrain, as shown by staining with anti-neurofilament antibody

Genotype: Pak4^tm1Amin/Pak4^tm1Amin
either: (involves: 129P2/OlaHsd * C57BL/6J) or (involves: 129S2/SvPas * C57BL/6J)

cardiovascular system phenotype

abnormal placenta vasculature
- at E10.5, vascular invasion fails after chorioallantoic fusion unlike in wild-type mice

growth/size/body region phenotype

decreased embryo size
- slightly at E9.5, markedly at E10.5

embryonic growth retardation
- severe at E10.5

integument phenotype

pallor
- slightly at E9.5, markedly at E10.5

cellular phenotype

increased apoptosis
- at E10.5, especially in the nervous system

decreased cell proliferation
- at E10.5, especially in the nervous system

embryo phenotype

abnormal vitelline vascular remodeling
- at E10.5, yolk sac vessels are dilated and contain fewer blood cells than in wild-type mice
- at E10.5, large vitelline vessels are absent with only the honeycomb-like plexus present

abnormal placenta labyrinth morphology
- at E10.5, the placenta labyrinth is compact with reduced thickness and fewer embryonic blood vessels compared to in wild-type mice

embryonic growth retardation
- severe at E10.5

pale yolk sac
- at E10.5

decreased embryo size
- slightly at E9.5, markedly at E10.5

abnormal placenta vasculature
- at E10.5, vascular invasion fails after chorioallantoic fusion unlike in wild-type mice

abnormal visceral yolk sac morphology
- at E10.5, yolk sacs are thicker and rougher than wild-type yolk sacs

pale placenta
- slightly at E9.5, markedly at E10.5

References

Qu J; Li X; Novitch BG; Zheng Y; Kohn M; Xie JM; Kozinn S; Bronson R; Beg AA; Minden A. 2003. PAK4 kinase is essential for embryonic viability and for proper neuronal development. Mol Cell Biol 23(20):7122-33PubMed: 14517283 MGI: J:85954

Additional - Pak4^tm1Amin related

Technical Support

CONTACT TECHNICAL SUPPORT

Genotyping Protocols
- Genotyping resources and troubleshooting
Breeding Considerations

When maintaining a live colony, heterozygotes may be bred to wildtype mice from the colony or to C57BL/6J inbred mice (Stock No. 000664). the donating investigator states that homozygous mice exhibit embryonic lethality by E11.5 due to heart defects.
- Additional Breeding and Husbandry Support
Citation
When using the B6.129S2-Pak4^tm1Amin/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #015829 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 Pak4<tm1Amin>

Related Products and Services

Frozen Mouse Embryo	B6.129S2-Pak4<tm1Amin>/J	$2595.00
Frozen Mouse Embryo	B6.129S2-Pak4<tm1Amin>/J	$2595.00
Frozen Mouse Embryo	B6.129S2-Pak4<tm1Amin>/J	$3373.50
Frozen Mouse Embryo	B6.129S2-Pak4<tm1Amin>/J	$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

Licensing Information

Phone: 207-288-6470

Email: TechTran@jax.org

Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Control Suggestions

Approximate Controls

Additional Information

Selected References

Pak4tm1Amin

Allele Symbol: Pak4tm1Amin

Disease Terms

Research Areas By Phenotype

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

Mammalian Phenotype Terms by Genotype

Genotype: Pak4tm1Amin/Pak4tm1Amineither: (involves: 129P2/OlaHsd * C57BL/6) or (involves: 129S2/SvPas * C57BL/6)

embryo phenotype

growth/size/body region phenotype

mortality/aging

muscle phenotype

nervous system phenotype

cardiovascular system phenotype

cellular phenotype

Genotype: Pak4tm1Amin/Pak4tm1Amineither: (involves: 129P2/OlaHsd * C57BL/6J) or (involves: 129S2/SvPas * C57BL/6J)

cardiovascular system phenotype

growth/size/body region phenotype

integument phenotype

cellular phenotype

embryo phenotype

References

Additional - Pak4tm1Amin 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

Licensing Information

Pak4^tm1Amin

Allele Symbol: Pak4^tm1Amin

Genotype: Pak4^tm1Amin/Pak4^tm1Amin
either: (involves: 129P2/OlaHsd * C57BL/6) or (involves: 129S2/SvPas * C57BL/6)

Genotype: Pak4^tm1Amin/Pak4^tm1Amin
either: (involves: 129P2/OlaHsd * C57BL/6J) or (involves: 129S2/SvPas * C57BL/6J)

Additional - Pak4^tm1Amin related