Mice that are homozygous for the targeted mutation exhibit impaired learning capacity in visible platform swimming water maze task and rotorod test, and abnormal eye blink conditioning response. Purkinje cell electrophysiology is abnormal. This mutant mouse strain may be useful in studies of learning, memory and neurophysiology.
Eric R Kandel, Columbia University
Mice that are homozygous for the targeted mutation are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities. No mRNA encoding the pore/S6 transmembrane domain is detected by in situ hybridization. No HCN1 protein is detected by Western blot analysis of membrane extracts from cerebellum, hippocampus or cortex. Mutant mice exhibit impaired learning capacity in visible platform swimming water maze task and rotorod test, and abnormal eye blink conditioning response. Purkinje cell electrophysiology is abnormal. This mutant mouse strain may be useful in studies of learning, memory and neurophysiology.
A loxP site flanked targeting vector containing neomycin resistance and thymidine kinase genes was utilized in the construction of this mutant. This selection cassette was inserted downstream from the exon encoding the p region and S6 transmembrane (pore-S6) domain, and another loxP site was inserted upstream of the same exon. The construct was electroporated into 129S/SvEv-derived MM13 embryonic stem (ES) cells (obtained from M. Mendelson) which were transiently transfected with a Cre recombinase vector to remove the selection cassette and the exon encoding the p region and S6 transmembrane (pore-S6) domain. Correctly targeted ES cells were injected into C57BL/6 blastocysts. (checking w DI?)
|Allele Name||targeted mutation 2, Eric R Kandel|
|Allele Type||Targeted (Null/Knockout)|
|Allele Synonym(s)||Hcn1-; HCN1 knockout|
|Gene Symbol and Name||Hcn1, hyperpolarization-activated, cyclic nucleotide-gated K+ 1|
|Strain of Origin||129S/SvEv|
|Molecular Note||A loxP site was inserted upstream of the pore-S6 domain containing exon. At the same time, a floxed neomycin cassette was introduced downstream. Cells in which transient Cre recombinase expression of properly targeted ES cells resulted in excision of both the neomycin gene and the pore-S6 exon were selected. Both mRNA and protein were shown to be absent in the brains of mice homozygous for this allele.|
|Mutations Made By|| |
Eric Kandel, Columbia University
The resulting chimeric animals were crossed to 129S/SvEv mice, and then backcrossed to C57BL/6 once. Heterozygotes were bred to generate homozygotes.
When using the B6;129-Hcn1tm2Kndl/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #005034 in your Materials and Methods section.