B6(Cg)-Tg(Thy1-Usp14*C114A)5-40Swi/J

Stock number: 036402
Product name: TgUsp14CA
Research areas: Cell Biology Research, Developmental Biology Research, Neurobiology Research

Description

Thy1-Usp14CA transgenic mice express a pan-neuronal specific mouse Usp14 (ubiquitin specific peptidase 14) gene modified to incorporate the C114A mutation, which alters the active site cysteine to alanine. Thy1-Usp14CA mice exhibit including impaired muscle development, motor endplate degeneration and impaired synaptic transmission at the neuromuscular junction.

Detailed description

Thy1-Usp14CA transgenic mice express a pan-neuronal (using the Thy1 promoter) specific mouse Usp14 (ubiquitin specific peptidase 14) gene modified to incorporate the C114A mutation, which alters the active site cysteine to alanine. Usp14 (ubiquitin specific peptidase 14) encodes a member of the ubiquitin-specific processing (UBP) family of proteases that functions as a deubiquitinating enzyme and is involved in recycling ubiquitin at the proteasome. Immunoblot analysis of Thy1-Usp14CA transgenics indicates that there is a robust increase in Usp14 expression in the spinal cord that is detectable by postnatal day 8. mRNA expression is not altered in the brain. USP14 abundance is not increased in non-neuronal tissues.

Mice hemizygous for the transgene are viable and fertile. The viability/fertility of homozygotes has not been assessed. Similar to the phenotype observed in ataxia-J mice (B6.Cg-Usp14^ax-J/J, Stock No. 000518), hemizygous Thy1-Usp14CA mice develop resting tremor and hind limb clasping by 2-3 weeks of age, decreased body mass by 12 weeks of age, reduced muscle mass and forelimb grip strength, and impaired ability on the elevated beam by 4 weeks of age. Mice exhibit a reduction in frequency of miniature endplate currents (MEPC) and an increase in the amplitude and decay constant of MEPCs. In addition, the neuromuscular junction (NMJ) structure develops focal swellings at the terminals and ultra-terminal sprouting by 4 weeks of age (57% and 56% of mice, respectively). The strain may be useful for studies of USP14 in neuromuscular disease.

Development

The Thy1-Usp14CA transgenic construct contains a modified full length mouse Usp14 cDNA (including the Usp14 Kozak sequence) under the control of the pan-neuronal Thy1(Thy1.2) promoter. The Usp14 sequence is modified to change the active site cysteine to an alanine (C114A) at position 114. The linearized construct was microinjected into the pronuclei of fertilized C57BL/6J. Founder line 5-40 was established and maintained on the C57BL/6J background. At some point, mice were crossed to mice carrying the Usp14^ax-J allele on the C57BL/6 background, the ax-J allele was later bred out. The strain has been backcrossed at least 13 times to C57BL/6J. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).

Allele

Symbol: Tg(Thy1-Usp14*C114A)5-40Swi
Name: transgene insertion 5-40, Scott Wilson
MGI accession ID: MGI:6715064
Generation method: Transgenic
Attributes: Inserted expressed sequence
Synonyms: TgUsp14CA
Marker symbol: Tg(Thy1-Usp14*C114A)5-40Swi
Marker name: transgene insertion 5-40, Scott Wilson
Chromosome: UN
Strain of origin: C57BL/6J
Molecular note: The transgenic construct contains a modified full length mouse Usp14 cDNA (including the Usp14 Kozak sequence) under the control of the pan-neuronal Thy1 (Thy1.2) promoter. The Usp14 sequence is modified to change the active site cysteine to an alanine (C114A) at position 114.