Overview

Also Known As:Thy1.2-PFN1^C71G/C71G/Prp-PFN1^C71G triple transgenic [or pt(h)] , Thy1.2-PFN1^C71G/Prp-PFN1^C71G double transgenic [or pt]

Thy1.2-PFN1^C71G/Prp-PFN1^C71G transgenic mice ("pt" or "pt(h)") express a C71G mutant human profilin 1 protein (with N-terminal V5 tag) predominantly in motor neurons and in both neurons and non-neuronal cells of the CNS. These mice exhibit motor neuron degeneration and abnormalities in cytoskeleton, proteasome, autophagy and stress granules. This is a model for studying the pathogenic role of PFN1 in familial amyotrophic lateral sclerosis (ALS or Lou Gehrig's disease).

Donating Investigator

Zuoshang Xu, University of Massachusetts Medical School

Genetic overview

Genetic Background	Generation
	N2+pN1F7 (2019-06-12 00:00:00)

Tg(Prnp-PFN1*C71G)22Zxu

Allele Type
Transgenic (Inserted expressed sequence, Humanized sequence)

Tg(Thy1-PFN1*C71G)67Zxu

Allele Type
Transgenic (Inserted expressed sequence, Humanized sequence)

View Genetics

Research Applications

Research Tools
Neurobiology Research
Cell Biology Research

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Base Price

Starting at:

$278.00 Domestic price for female 4-week

556.00 Domestic price for breeder pair

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Details

Detailed Description

These mice harbor two transgenes, Prp-PFN1^C71G and Thy1.2-PFN1^C71G, each expressing a C71G mutant human profilin 1 protein with N-terminal V5 tag (V5-PFN1^C71G) under direction of the mouse prion protein promoter or the human Thy1.2 promoter.

The Prp-PFN1^C71G transgene has V5-PFN1^C71G expression predominantly in the CNS (in both neurons and non-neuronal cells), with very low expression in the periphery. Specifically, expression is highest in the spinal cord, brainstem and forebrain, lower levels in cerebellum and kidney, and at undetectable levels in other organs.

The Thy1.2-PFN1^C71G transgene drives V5-PFN1^C71G expression predominantly in motor neurons. Specifically, expression is highest in the spinal cord and brainstem, followed by the forebrain. Little if any is expressed in the cerebellum and the peripheral organs.

When hemizygous for both transgenes, they are referred to as Thy1.2-PFN1^C71G/Prp-PFN1^C71G double transgenic mice (and were previously called "pt" mice).

When homozygous for Thy1.2-PFN1^C71G and hemizygous for Prp-PFN1^C71G are referred to as
Thy1.2-PFN1^C71G/C71G/Prp-PFN1^C71G triple transgenic mice (and were previously called "pt(h)" mice).

Furthermore, mice harboring only the Thy1.2-PFN1^C71G transgene are referred to as Thy1.2-PFN1^C71G (previously called "t") when hemizygous, and Thy1.2-PFN1^C71G/C71G (previously called "t(h)") when homozygous.

Similarly, mice harboring only the Prp-PFN1^C71G transgene are referred to as Prp-PFN1^C71G (previously called "p") when hemizygous, and Prp-PFN1^C71G/C71G (previously called "p(h)") when homozygous.

In order of most-to-least motor neuron disease severity: "pt(h)" : "t(h)" : "pt" : "t" : "p".

The "pt(h)" and "t(h)" mice have similar disease onset age (~125d), but "pt(h)" mice develop paralysis faster than "t(h)" mice (~60d after onset compared to ~170d after onset). While "t(h)" and "pt" mice have similar overall life expectancy (~250-350d), the "pt" mice have a much later disease onset (~250d) with rapid development of paralysis (~10d after onset).

The "pt(h)" genotype is most severely affected, with mice exhibiting motor neuron degeneration and abnormalities in cytoskeleton, proteasome, autophagy and stress granules.
Specifically, beginning at 3-4 months of age, "pt(h)" exhibit progressive loss of motor capabilities, with progressive loss of motor and sensory axons. Cytoskeleton defects in motor neurons include mutant PFN1 aggregation and reduced neurofilament L expression.
Alterations in autophagy pathway are evident throughout spinal cord, particularly in motor neurons.
At the paralysis disease stage (end stage), there is significant microgliosis, astrogliosis, motor neuron loss, elevated ubiquitin levels in the spinal cord, increased stress granules in the spinal motor neurons

Both "pt" and "pt(h)" mice are viable and fertile. The donating investigator reports that "pt(h)" mice breed normally, and they maintain their double transgenic colony by breeding "t(h)" females with "pt(h)" males.

Mice singly hemizygous for the Prp-PFN1^C71G transgene ("p") express V5-PFN1^C71G in both neurons and glia, but the expression levels are low. The "p" genotype does not manifest motor neuron degeneration symptoms during their lifespan. The donating investigator also reports they were not able to obtain homozygous Prp-PFN1^C71G animals ("p(h)").

Development

These mice harbor two transgenes, Prp-PFN1^C71G and Thy1.2-PFN1^C71G, each designed by Drs. Zuoshang Xu and John E. Landers (University of Massachusetts Medical School). The PFN1^C71G protein (V5-PFN1^C71G) encoded by each transgene is a human profilin 1 cDNA sequence (PFN1; 423 bp / 141 aa) harboring the C71G mutation associated with amyotrophic lateral sclerosis (ALS), with a V5-tag (45 bp) fused to its N-terminus.

The Prp-PFN1^C71G transgene has the V5-PFN1^C71G sequence inserted between exon 2 and exon 3 of mouse prion protein (PrP or Prnp) gene at a unique XhoI site in the MoPrP.Xho plasmid vector. The resulting 11,855 bp Prp-V5-PFN1 construct was microinjected into FVB/NJ embryos. Transgenic founder male 22 was bred with FVB/NJ females for germline transmission, and founder line 22 was found to have expression predominantly in the CNS. The integration site(s) and copy number of the Prp-PFN1^C71G transgene is not known to date (February 2016). The Prp-PFN1^C71G line 22 colony was maintained by breeding hemizygotes with FVB/NJ mice for at least one more generation, prior to using as below.

The Thy1.2-PFN1^C71G transgene has the V5-PFN1^C71G sequence inserted downstream of the modified ~6.5 kbp mouse Thy1.2 promoter sequences (Thy1; thymus cell antigen 1, theta). The resulting 1,171 bp Thy1-V5-PFN1 construct was microinjected into FVB/NJ embryos. Transgenic founder female 67 was bred with FVB/NJ males for germline transmission, and founder line 67 was found to have expression predominantly in the CNS. The integration site(s) and copy number of the Thy1.2-PFN1^C71G transgene is not known to date (February 2016). The Thy1.2-PFN1^C71G line 67 colony was maintained by breeding hemizygotes with FVB/NJ mice for at least one more generation, prior to using as below.

To generate the Prp-PFN1^C71G Thy1.2-PFN1^C71G mice, the two individual mouse lines were bred together. The donating investigator then maintained the colony by breeding females homozygous for Thy1.2-PFN1^C71G ("t(h)" females) with males hemizygous for PrP-PFN1^C71G and homozygous for Thy1.2-PFN1^C71G ("pt(h)" males). In 2016, albino males harboring both transgenes were sent to The Jackson Laboratory Repository. Upon arrival, sperm was cryopreserved. To generate our live colony, an aliquot of the frozen sperm was used to fertilize oocytes from FVB/NJ mice (Stock No. 001800).

Expression Data

Expressed Gene	PFN1, profilin 1, human
Site of Expression
Expressed Gene	PFN1, profilin 1, human
Site of Expression

Control Suggestions

Noncarrier

Approximate Controls

001800 FVB/NJ

Additional Information

Considerations for Choosing Controls

Selected References

Yang C; Danielson EW; Qiao T; Metterville J; Brown RH Jr; Landers JE; Xu Z. 2016. Mutant PFN1 causes ALS phenotypes and progressive motor neuron degeneration in mice by a gain of toxicity. Proc Natl Acad Sci U S A 113(41):E6209-E6218PubMed: 27681617 MGI: J:235427

View All References

Genetics

Tg(Prnp-PFN1*C71G)22Zxu

Allele Symbol: Tg(Prnp-PFN1*C71G)22Zxu

Allele Name	transgene insertion 22, Zuoshang Xu
Allele Type	Transgenic (Inserted expressed sequence, Humanized sequence)
Allele Synonym(s)	Prp-PFN1^C71G
Gene Symbol and Name	Tg(Prnp-PFN1*C71G)22Zxu, transgene insertion 22, Zuoshang Xu
Gene Synonym(s)
Promoter	Prnp, prion protein, mouse, laboratory
Expressed Gene	PFN1, profilin 1, human
Strain of Origin	FVB/N
Chromosome	UN
Molecular Note	The transgenic construct contains a mutant human profilin 1 cDNA sequence with a V5 tag (45 bp) fused to the N-terminus and inserted between exon 2 and exon 3 of mouse prion protein gene at a unique XhoI site in the MoPrP.Xho plasmid vector. PFN1 harbors the C71G mutation associated with amyotrophic lateral sclerosis (ALS).

Tg(Thy1-PFN1*C71G)67Zxu

Allele Symbol: Tg(Thy1-PFN1*C71G)67Zxu

Allele Name	transgene insertion 67, Zuoshang Xu
Allele Type	Transgenic (Inserted expressed sequence, Humanized sequence)
Allele Synonym(s)	Thy1.2-PFN1^C71G
Gene Symbol and Name	Tg(Thy1-PFN1*C71G)67Zxu, transgene insertion 67, Zuoshang Xu
Gene Synonym(s)
Promoter	Thy1, thymus cell antigen 1, theta, mouse, laboratory
Expressed Gene	PFN1, profilin 1, human
Strain of Origin	Not Specified
Chromosome	UN
Molecular Note	The transgenic construct contains a mutant human profilin 1 cDNA sequence with a V5 tag (45 bp) fused to the N-terminus and inserted downstream of the modified ~6.5 kbp mouse Thy1.2 promoter sequences. PFN1 harbors the C71G mutation associated with amyotrophic lateral sclerosis (ALS).

Disease/Phenotype

Disease Terms

Characteristics of this human disease are associated with transgenes and other mutation types in the mouse.

amyotrophic lateral sclerosis type 18

Research Areas By Phenotype

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

Research Tools
- Cell Biology Research
- Neurobiology Research
Neurobiology Research
- Neurodegeneration
- Neuromuscular defects
- Amyotrophic Lateral Sclerosis (ALS)
- Ataxia (Movement) Defects
Cell Biology Research
- Protein Processing
  - degradation
- Cell Cycle Regulation

Mammalian Phenotype Terms by Genotype

The following phenotype relates to a compound genotype created using this strain

Genotype: Tg(Prnp-PFN1C71G)22Zxu/0 Tg(Thy1-PFN1C71G)67Zxu/0
involves: FVB/N

behavior/neurological phenotype

paralysis
- paralysis at 305 +/-34 days

mortality/aging

premature death
- due to paralysis

muscle phenotype

progressive muscle weakness
- muscle weakness beginning at an average age of 295 days

Genotype: Tg(Prnp-PFN1C71G)22Zxu/0 Tg(Thy1-PFN1C71G)67Zxu/Tg(Thy1-PFN1*C71G)67Zxu
involves: FVB/N

behavior/neurological phenotype

decreased grip strength
- reduced grip strength beginning at 121-150 days

decreased vertical activity
- impaired vertical behavior beginning at 121-150 days

paralysis
- paralysis at 211 +/-25 days

nervous system phenotype

abnormal motor neuron innervation pattern
- increased muscle denervation

abnormal motor neuron morphology
- motor neurons have reduced neurofilaments, lack a well-spread filament network and exhibit a circular distribution of neurofilaments around the cell periphery

astrocytosis
- astrogliosis is increased in ventral horn of spinal cord at the end stage of disease

axon degeneration
- axon degeneration is observed in the dorsal root, but progression lags behind ventral root axons
- axon degeneration is observed in white matter of spinal cord; degeneration is mild in lateral column and more severe in ventral column
- axon loss is progressive
- increased number of degenerating axons in the ventral root in 100-120 day old mice as compared to controls
- little axon degeneration is observed in the cortical spinal track in the dorsal column

decreased motor neuron number
- motor neuron number is reduced in lumbar spinal cord at the end stage of disease (170-226 days)
- motor neuron number is reduced in the ventral horn of cervical spinal cord beginning at 132-154 days of age

microgliosis
- microgliosis is increased in ventral horn of spinal cord at the end stage of disease

motor neuron degeneration

neurodegeneration
- increased cellulation (and GFAP staining) in medulla
- neuronal loss is not observed in cerebral cortex, hippocampus or cerebellum

neuronal cytoplasmic inclusions
- 1-2% of motor neurons form large aggregates
- motor neurons exhibit widespread small particulates in the cytoplasm and neuronal processe
- p62/SQSTM1 staining is present as small particulates in cytoplasm of motor neurons
- protein aggregates are visible by day 127 and gradually accumulate until death
- strong ubiquitin staining is present in motor neurons and some neuropils

cellular phenotype

abnormal cell cytoskeleton morphology
- motor neurons have reduced neurofilaments, lack a well-spread filament network and exhibit a circular distribution of neurofilaments around the cell periphery

growth/size/body region phenotype

decreased body weight
- reduced body weight beginning at 121-150 days

hematopoietic system phenotype

microgliosis
- microgliosis is increased in ventral horn of spinal cord at the end stage of disease

immune system phenotype

microgliosis
- microgliosis is increased in ventral horn of spinal cord at the end stage of disease

mortality/aging

premature death
- due to paralysis

muscle phenotype

increased variability of skeletal muscle fiber size
- clusters of oxidative (SDH-strong) and weak oxidative (SDH-weak) fibers as compared to interspersed distribution found in control

progressive muscle weakness
- 10% of mice between 100-120 days old exhibit slight weakness (foot dragging)
- 100% of mice between 141-170 days exhibit slight weakness, progressing to weakness (leg dragging)
- 70% of mice between 121-140 days old exhibit slight weakness
- beyond 170 days, all mice progress from partial paralysis to paralysis in both fore and hind limbs
- impaired rotorod performance beginning at 121-150 days

skeletal muscle fiber atrophy

References

Yang C; Danielson EW; Qiao T; Metterville J; Brown RH Jr; Landers JE; Xu Z. 2016. Mutant PFN1 causes ALS phenotypes and progressive motor neuron degeneration in mice by a gain of toxicity. Proc Natl Acad Sci U S A 113(41):E6209-E6218PubMed: 27681617 MGI: J:235427

Additional - Tg(Prnp-PFN1*C71G)22Zxu related

Additional - Tg(Thy1-PFN1*C71G)67Zxu related

Technical Support

CONTACT TECHNICAL SUPPORT

Genotyping Protocols
- Standard PCR:Tg(Prnp-PFN1)34Zxu-alt1
- Standard PCR:Tg(Thy1-PFN1*)67Zxu-Alternate 2
- Genotyping resources and troubleshooting
Breeding Considerations

Prp-PFN1^C71G transgenic mice were referred to as "p" or "p(h)" when hemizygous or homozygous, respectively.

Similarly, Thy1.2-PFN1^C71G transgenic mice were referred to as "t" or "t(h)" when hemizygous or homozygous, respectively.

When hemizygous for both transgenes, they are referred to as Thy1.2-PFN1^C71G/Prp-PFN1^C71G double transgenic mice (and were previously called "pt" mice).

When homozygous for Thy1.2-PFN1^C71G and hemizygous for Prp-PFN1^C71G are referred to as Thy1.2-PFN1^C71G/C71G/Prp-PFN1^C71G triple transgenic mice (and were previously called "pt(h)" mice).

The donating investigator reports that "pt(h)" mice breed normally. They also report that breeding "t(h)" females with "pt(h)" males results in 50% "t(h)" offspring and 50% "pt(h)" offspring.

When maintaining our live colony at The Jackson Laboratory Repository, we will breed mice wildtype (noncarrier) for Prp-PFN1^C71G and hemizygous for Thy1.2-PFN1^C71G with mice hemizygous for Prp-PFN1^C71G and homozygous for Thy1.2-PFN1^C71G. That is, we will breed "t" mice with "pt(h)" mice.
- Additional Breeding and Husbandry Support
Mating System
- wildtype (noncarrier) for Prp-PFN1C71G and homozygous for Thy1.2-PFN1C71G X hemizygous for Prp-PFN1C71G and homozygous for Thy1.2-PFN1C71G
Citation
When using the Thy1.2-PFN1^C71G/C71G/Prp-PFN1^C71G triple transgenic [or pt(h)] , Thy1.2-PFN1^C71G/Prp-PFN1^C71G double transgenic [or pt] mouse strain in a publication, please cite the originating article(s) and include JAX stock #028608 in your Materials and Methods section.

Animal Health Reports

Facility Barrier Level Descriptions

AX11 (Maximum)

Pricing & Availability

Available

Domestic
International

Live Mouse

Age

Genotype

Price

weeks

Breeder Pair

Sex

Genotype

Price

Female
Male

Cryorecovery - Pricing

Service/Product	Description	Price
	Hemizygous for Tg(Prnp-PFN1C71G)22Zxu Hemizygous for Tg(Thy1-PFN1C71G)67Zxu or Hemizygous for Tg(Prnp-PFN1C71G)22Zxu non carrier for Tg(Thy1-PFN1C71G)67Zxu

Related Products and Services

Frozen Mouse Embryo	FVB/N-Tg(Prnp-PFN1C71G)22Zxu Tg(Thy1-PFN1C71G)67Zxu/J	$2595.00
Frozen Mouse Embryo	FVB/N-Tg(Prnp-PFN1C71G)22Zxu Tg(Thy1-PFN1C71G)67Zxu/J	$2595.00
Frozen Mouse Embryo	FVB/N-Tg(Prnp-PFN1C71G)22Zxu Tg(Thy1-PFN1C71G)67Zxu/J	$3373.50
Frozen Mouse Embryo	FVB/N-Tg(Prnp-PFN1C71G)22Zxu Tg(Thy1-PFN1C71G)67Zxu/J	$3373.50

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Also Known As:Thy1.2-PFN1C71G/C71G/Prp-PFN1C71G triple transgenic [or pt(h)] , Thy1.2-PFN1C71G/Prp-PFN1C71G double transgenic [or pt]

Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Expression Data

Control Suggestions

Approximate Controls

Additional Information

Selected References

Tg(Prnp-PFN1*C71G)22Zxu

Allele Symbol: Tg(Prnp-PFN1*C71G)22Zxu

Tg(Thy1-PFN1*C71G)67Zxu

Allele Symbol: Tg(Thy1-PFN1*C71G)67Zxu

Disease Terms

Characteristics of this human disease are associated with transgenes and other mutation types in the mouse.

Research Areas By Phenotype

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

Mammalian Phenotype Terms by Genotype

Genotype: Tg(Prnp-PFN1*C71G)22Zxu/0 Tg(Thy1-PFN1*C71G)67Zxu/0involves: FVB/N

behavior/neurological phenotype

mortality/aging

muscle phenotype

Genotype: Tg(Prnp-PFN1*C71G)22Zxu/0 Tg(Thy1-PFN1*C71G)67Zxu/Tg(Thy1-PFN1*C71G)67Zxuinvolves: FVB/N

behavior/neurological phenotype

nervous system phenotype

cellular phenotype

growth/size/body region phenotype

hematopoietic system phenotype

immune system phenotype

mortality/aging

muscle phenotype

References

Additional - Tg(Prnp-PFN1*C71G)22Zxu related

Additional - Tg(Thy1-PFN1*C71G)67Zxu related

Genotyping Protocols

Breeding Considerations

Mating System

Citation

Animal Health Reports

Live Mouse

Breeder Pair

Cryorecovery - Pricing

Related Products and Services

Payment Terms and Conditions

The Jackson Laboratory's Genotype Promise

Terms Of Use

Licensing Information

JAX® Mice, Products & Services Conditions of Use

No Warranty

Credit for PRODUCTS or SERVICES

Animal Care and Use for SERVICES

No Liability

Also Known As:Thy1.2-PFN1^C71G/C71G/Prp-PFN1^C71G triple transgenic [or pt(h)] , Thy1.2-PFN1^C71G/Prp-PFN1^C71G double transgenic [or pt]

Genotype: Tg(Prnp-PFN1C71G)22Zxu/0 Tg(Thy1-PFN1C71G)67Zxu/0
involves: FVB/N

Genotype: Tg(Prnp-PFN1C71G)22Zxu/0 Tg(Thy1-PFN1C71G)67Zxu/Tg(Thy1-PFN1*C71G)67Zxu
involves: FVB/N