TITLE:
024414 B6.129(SJL)-Grip1tm1Rlh
024415 B6.129S6(Cg)-Wwc1tm1.1Rlh
024417 B6.129S6(SJL)-Prkcz tm1.1Rlh
024418 B6.129S6-Gria1tm7Rlh
024419 B6.129(Cg)-Gria2tm2Rlh
024420 B6.129(Cg)-Gria1tm4Rlh
CASE NUMBER: C13635
ABSTRACT
Information are from Jacksons Lab website (https://www.jax.org/mouse-search?searchTerm=Richard%20Huganir)
Stock No. 024414 B6.129(SJL)-Grip1tm1Rlh
Description
Grip1 (glutamate receptor interacting protein 1) binds GluR2 (Gria2)-containing α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors and couples them to structural and signaling complexes in neurons. Deletion of both GRIP1 and GRIP2 blocks long-term synaptic depression (LTD) in primary cultures of mouse cerebellar neurons in a model system of synaptic plasticity.
The first PDZ (postsynaptic density protein-95 (PSD-95)/Drosophila disc large tumor suppressor (Dlg1)/zona occludens-1 protein) domain of the Grip1 gene is flanked by loxP sites in this conditional targeted mutant strain. Expression of Cre recombinase results in the deletion of the first PDZ1 exon, causing a frameshift mutation that introduces an immediate stop codon in the GRIP1 transcript.
Development
Two exons encode the first PDZ (postsynaptic density protein-95 (PSD-95)/Drosophila disc large tumor suppressor (Dlg1)/zona occludens-1 protein) domain of the targeted gene. An Frt-flanked PGK neomycin cassette (in reverse transcriptional orientation) followed by a loxP site was introduced to the intron located between the two exons. An additional loxP site was placed 5' of the first PDZ exon. The targeted mutation was created through homologous recombination in (129X1/SvJ x 129S1/Sv)F1- Kitl+-derived R1 embryonic stem (ES) cells. Resultant chimeric animals were bred to C57BL/6 mice. Heterozygotes were then bred to C57BL/6 background animals expressing FLP recombinase from a human ACTB promoter (Tg(ACTFLPe)#Dym) to delete the neomycin cassette. This strain was backcrossed to C57BL/6 for 10 generations by the donating laboratory.
Stock No. 024415 B6.129S6(Cg)-Wwc1tm1.1Rlh
Description
Mutations in the WWC1 (WW, C2 and coiled-coil domain containing 1; also called KIBRA) gene have been associated with human memory performance.
This strain carries a targeted knockout of the mouse Wwc1 gene. Western blot analysis confirmed the absence of protein expression in the brain. Adult homozygous mice demonstrate impaired long-term depression and long-term potentiation at hippocampal Schaffer collateral-CA1 synapses. They also show severe deficits in contextual fear learning and memory. WWC1/KIBRA directly binds PICK1 in vitro and in vivo and interacts with GluA1 (GRIA1), GluA2 (GRIA2), and several other synaptic proteins in an in vivo protein complex. Evidence suggests that the gene plays and important role in regulating AMPAR (α-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate receptor) trafficking underlying synaptic plasticity and learning.
Development
A loxP site was introduced upstream of exon 4 and an FRT-flanked PGK-neomycin cassette followed by a loxP site was placed downstream of exon 5. The targeting vector was introduced to 129S6/SvEvTac-derived MC1 embryonic stem (ES) cells. Resultant germline mice were crossed with a CMV-Cre strain on a C57BL/6 background to excise the floxed region, resulting in an out-of-frame mutation. This strain was backcrossed to C57BL/6 for 10 generations by the donating laboratory.
Stock No. 024417 B6.129S6(SJL)-Prkcz tm1.1Rlh
Description
Sustained activation of the brain-specific protein kinase C (PKC) isoform protein kinase M-ζ (Prkcz, protein kinase C, zeta; also called PKM-ζ) has been reported to be necessary for both long-term potentiation (LTP) maintenance and long-term memory. PKM-zeta; is transcribed from an alternate internal promoter within the Prkcz gene encoding only the catalytic domain, rendering it free from pseudosubstrate auto-inhibition and PKC-ζ regulatory signal transduction pathways.
In this conditional mutant allele, exon 11 of the mouse Prkcz gene (encoding the catalytic domain) is flanked by loxP sites, enabling directed, Cre-mediated knockout of the PKM-&zeta isoform (referred to as PKC-ζ/PKM-ζ KO)
Knockout mice derived from crosses with CMV-Cre mice show normal synaptic transmission and LATP at Schaffer collateral-CA1 synapses, and have no deficits in several hippocampal-dependent learning and memory tasks.
Development
A loxP site was introduced upstream of exon 11 and a neomycin resistance cassette flanked by FRT sites (in reverse transcriptional orientation) followed by a loxP site were placed downstream of exon 11. The mutation was created by homologous recombination in 129S6/SvEvTac-derived MC1 embryonic stem (ES) cells. The neomycin cassette was removed from resultant mice through crosses with an a human ACTB Flp strain (Tg(ACTFLPe)#Dym) on a C57BL/6 background. This strain was backcrossed to C57BL/6 for 10 generations by the donating laboratory.
Stock No. 024418 B6.129S6-Gria1tm7Rlh
Description
Phosphorylation of Gria1 (glutamate receptor, ionotropic, AMPA1 (alpha 1); also known as GluA1) has been proposed to regulate receptor trafficking and synaptic transmission and plasticity.
These mice carry S831D and S845D amino acid substitutions in the mouse Gria1 gene that are phosphomimetic (mimic a phosphorylated protein). Mice show normal total and surface expression and subcellular localization of GLUA1 as well as intact basal synaptic transmission. Unlike wildtype animals, mutant mice show long-term potentiation (LTP) induced with a weak 10-Hz stimulation. Evidence indicates that phosphorylation of GLUA1 at S831 and S845 is sufficient to lower the threshold for LTP induction, increasing the probability of synaptic plasticity.
Development
S831D (TCC->GAT) and S845D (AGC->GAC) serine to aspartate mutations were introduced to GluA1 (Gria1) cDNA using QuikChange site-directed mutagenesis (Stratagene). The mutations were introduced to 129S6/SvEvTac-derived MC1 embryonic stem (ES) cells by homologous recombination. This strain was backcrossed to C57BL/6 for 10 generations by the donating laboratory.
Stock No. 024419 B6.129(Cg)-Gria2tm2Rlh
Description
Cerebellar long-term depression (LTD) is a major form of synaptic plasticity that is thought to be critical for certain types of motor learning. Phosphorylation of GluR2 (Gria2, glutamate receptor, ionotropic, AMPA2 (alpha 2)) on serine-880 has been suggested to contribute to the endocytic removal of postsynaptic AMPA receptors during LTD.
In this mutant strain, a lysine (K) to alanine (A) mutation was introduced to the consensus recognition motif for protein kinase C (PKC) at amino acid 882 to prevent PKCα-mediated phosphorylation at serine 880 (S880) of the GRIA2 protein. Western blots performed with a phospho-specific S880 antibody reveal that basal S880 phosphorylation is intact at levels similar to those observed in wildtype littermates, but PKC-mediated S880 phosphorylation is abolished. The mutation does not interfere with binding to the PDZ domain-containing proteins PICKI and GRIPI. Cerebellar architecture and Purkinje cell morphology appear grossly normal by Nissl and Golgi staining. Cerebellar LTD is absent. Mice are viable and without any observable developmental defects.
Development
A lysine (K) to alanine (A) change at amino acid 882 was created in exon 15 (encoding transmembrane domain 4 and the C terminus) of Gria2. This corresponds to an AAA ATT TAG to GCC ATA TGA sequence change (also introducing a BslI site). A loxP-flanked neomycin cassette was introduced to the intron 14 in reverse transcriptional orientation. Linearized targeting vector was electroporated into (129X1/SvJ x 129S1/Sv)F1-Kitl+-derived R1 embryonic stem (ES) cells. Resultant chimeric mice were checked for germline transmission and the neomycin cassette was excise by breeding to CMV-Cre mice. Cre was bred out of the line by backcrossing to C57BL/6 for 10 generations.
Stock No. 024420 B6.129(Cg)-Gria1tm4Rlh
Description
Phosphorylation of various AMPA receptor subunits, including Gria1 (GluR1), can alter synaptic transmission and plasticity at excitatory glutamatergic synapses in the central nervous system.
These GluR1 "penta" phosphomutant mice express five alanine substitutions (S831A, T838A, S839A, T840A, and S845A) that were introduced to the mouse Gria1 (glutamate receptor, ionotropic, AMPA1 (alpha 1)) gene, thus blocking T840 phosphorylation and preventing phosphorylation from other upstream sites. Western blots indicate that GRIA1 protein is still expressed in homozygotes. Hippocampal cytoarchitecture is normal as shown by Nissl stain, and protein distribution is normal as shown by immunohistochemistry. Long-term potentiation (LTP) induced by theta burst stimulation (TBS) is reduced in adult homozygotes, and although long-term depression (LTD) induced by a paired-pulse 1 Hz protocol is absent in adult animals, it is present in young animals. Mutant mice do not show any gross abnormalities in anatomy and behavior.
Development
A neomycin resistance cassette flanked by loxP sites was inserted to the intron upstream of the last coding exon. Five alanine substitutions (S831A, T838A, S839A, T840A, and S845A) were introduced to phosphorylation sites by directed PCR. The mutations were introduced through homologous recombination in (129X1/SvJ x 129S1/Sv)F1- Kitl+-derived R1 embryonic stem (ES) cells. After germline transmission was confirmed, the neomycin cassette was excised through crosses with a CMV-Cre strain on a mixed C57BL/6-129 genetic background. This strain was backcrossed to C57BL/6 for 12 generations by the donating lab.
INVENTORS: Richard L. Huganir
ASSOCIATED PUBLICATIONS:
Stock No. 024414: Takamiya K; Mao L; Huganir RL; Linden DJ. 2008. The glutamate receptor-interacting protein family of GluR2-binding proteins is required for long-term synaptic depression expression in cerebellar Purkinje cells. J Neurosci 28(22):5752-5. [PubMed: 18509036] [MGI Ref ID J:136388]
Mao L; Takamiya K; Thomas G; Lin DT; Huganir RL. 2010. GRIP1 and 2 regulate activity-dependent AMPA receptor recycling via exocyst complex interactions. Proc Natl Acad Sci U S A 107(44):19038-43. [PubMed: 20956289] [MGI Ref ID J:166238]
Mejias R; Adamczyk A; Anggono V; Niranjan T; Thomas GM; Sharma K; Skinner C; Schwartz CE; Stevenson RE; Fallin MD; Kaufmann W; Pletnikov M; Valle D; Huganir RL; Wang T. 2011. Gain-of-function glutamate receptor interacting protein 1 variants alter GluA2 recycling and surface distribution in patients with autism. Proc Natl Acad Sci U S A 108(12):4920-5. [PubMed: 21383172] [MGI Ref ID J:170091]
Stock No. 024415: Makuch L; Volk L; Anggono V; Johnson RC; Yu Y; Duning K; Kremerskothen J; Xia J; Takamiya K; Huganir RL. 2011. Regulation of AMPA receptor function by the human memory-associated gene KIBRA. Neuron 71(6):1022-9. [PubMed: 21943600] [MGI Ref ID J:178552]
Stock No. 024417: Volk LJ; Bachman JL; Johnson R; Yu Y; Huganir RL. 2013. PKM-zeta is not required for hippocampal synaptic plasticity, learning and memory. Nature 493(7432):420-3. [PubMed: 23283174] [MGI Ref ID J:194557]
Stock No. 024418: Makino Y; Johnson RC; Yu Y; Takamiya K; Huganir RL. 2011. Enhanced synaptic plasticity in mice with phosphomimetic mutation of the GluA1 AMPA receptor. Proc Natl Acad Sci U S A 108(20):8450-5. [PubMed: 21536866] [MGI Ref ID J:172128]
Stock No. 024419: Steinberg JP; Takamiya K; Shen Y; Xia J; Rubio ME; Yu S; Jin W; Thomas GM; Linden DJ; Huganir RL. 2006. Targeted in vivo mutations of the AMPA receptor subunit GluR2 and its interacting protein PICK1 eliminate cerebellar long-term depression. Neuron 49(6):845-60. [PubMed: 16543133] [MGI Ref ID J:107573]
Schonewille M; Gao Z; Boele HJ; Veloz MF; Amerika WE; Simek AA; De Jeu MT; Steinberg JP; Takamiya K; Hoebeek FE; Linden DJ; Huganir RL; De Zeeuw CI. 2011. Reevaluating the role of LTD in cerebellar motor learning. Neuron 70(1):43-50. [PubMed: 21482355] [MGI Ref ID J:174710]
Stock No. 024420: Lee HK; Takamiya K; Kameyama K; He K; Yu S; Rossetti L; Wilen D; Huganir RL. 2007. Identification and characterization of a novel phosphorylation site on the GluR1 subunit of AMPA receptors. Mol Cell Neurosci 36(1):86-94. [PubMed: 17689977] [MGI Ref ID J:126784]
CONTACT INFORMATION
Carole Burns
Senior Licensing Associate
JHTV
410-516-6492
cburns21@jhu.edu
