B6.Cg-Tg(Pnkd*A7V*A9V,-DsRed)704Ljp/J

Stock number: 022146
Research areas: Metabolism Research, Neurobiology Research

Description

This transgenic strain, mut Tg Pknd line 704, expresses an estimated 2 copies of the long isoform of the paroxysmal nonkinesigenic dyskinesia (Pnkd) gene mutated to include the A7V and A9V substitutions found in human PNKD patients. Following i.p. administration of ethanol or caffeine, mice exhibit nigrostriatal neurotransmission deficits and dyskinesia.

Detailed description

The autosomal dominant episodic movement disorder, paroxysmal nonkinesigenic dyskinesia (PKND), is associated with two valine to alanine substitutions in the uncharacterized human gene, PNKD. This transgenic strain, mut Tg (line 704), expresses the long isoform of the Pnkd gene and the A7V and A9V substitutions. The long isoform of PNKD is expressed only in the central nervous system. In humans, PNKD movement disorder is observed following stress, caffeine or alcohol intake. Mice carrying the transgene are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities. Following i.p. administration of ethanol or caffeine, transgenic mice exhibit nigrostriatal neurotransmission deficits, reduced extracellular dopamine levels in the striatum, an increased percentage of striatal dopamine release, and dyskinesia characterized by severe axial stiffness and abnormal limb movements. Non-induced mice exhibit lower glutathione levels in the frontal cortex. This mutant mouse strain may be useful in studies of autosomal dominant episodic movement disorder (PKND) and cellular redox regulation. No discernible phenotypic differences have been observed between transgenic line 671 (see Stock No. 018410) which carries an estimated single copy of the transgene and line 704 which carries approximately 2 copies. A DsRed insertion is believed to be non-functional.

Development

The bacterial artificial chromosome (BAC) RP24-112K19 containing the entire paroxysmal nonkinesiogenic dyskinesia (Pnkd) gene and 62 Kb of flanking sequence 5' to the AATG start site was altered to insert the mutations associated with human PNKD - valine to alanine substitutions at amino acid positions 7 (A7V) and 9 (A9V). An internal ribosomal entry site (IRES) followed by an enhanced red fluorescent protein (DsRED) also was inserted into the 3' UTR of Pnkd. This modified BAC was microinjected into the pronucleus of C57BL/6XSJL F1 oocytes. Founder mice (line 704) were established and found to have an estimated 2 copies of the transgene. These mut-Tg Pnkd transgenic mice were subsequently backcrossed to C57BL/6J for at least 10 generations prior to arrival at The Jackson Laboratory. Upon arrival, mice were bred to C57BL/6J (Stock No. 000664) for at least one generation to establish the colony. The DsRED insertion is believed to be non-functional.

Allele

Symbol: Tg(Pnkd*A7V*A9V,-DsRed)704Ljp
Name: transgene insertion 704, Louis J Ptacek
MGI accession ID: MGI:5466584
Generation method: Transgenic
Attributes: Inserted expressed sequence
Marker symbol: Tg(Pnkd*A7V*A9V,-DsRed)704Ljp
Marker name: transgene insertion 704, Louis J Ptacek
Chromosome: UN
Strain of origin: (C57BL/6 x SJL)F1
Expressed genes: RFP,Red Fluorescent Protein,coral
Molecular note: The transgene contains a BAC with the nucleotide substitutions that lead to the amino acid substitutions of valine for alanine at positions 7 and 9. An IRES/DsRed was inserted into the 3'UTR. This transgene is predicted to produce only the long isoform. Lines 671, 676, and 704 were generated.