These mice carry a targeted mutation with exons 14 through 17 of the Cacna1f (calcium channel, voltage-dependent, alpha 1F subunit) gene deleted. This mutant mouse strain may be useful in studies of calcium channelopathies, incomplete congenital stationary night blindness, photoreceptor electrophysiology, and the role of the Cav1.4 channel in survival of naive T cells.
L-type voltage dependent calcium channel activity is one of the primary ways calcium ions enter the cell from the extracellular space and has a critical role in intracellular signaling. The alpha 1F subunit makes up the central pore forming portion of the L-type voltage dependent calcium channel, Cav1.4. Genetic defects in Cav1.4 can cause human incomplete congenital stationary night blindness. Female mice that are homozygous and male mice that are hemizygous for the targeted mutation are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities. A lower molecular weight gene product (protein) is detected by Western blot analysis of retinal tissue. Immunohistochemical examination of adult retina from homozygotes reveals diminished and diffuse staining for Cacna1s (calcium channel, voltage-dependent, L type, alpha 1S subunit), mGluR6 (glutamate receptor 6), Serca2 (ATPase, Ca++ transporting, cardiac muscle, slow twitch 2), and photoreceptor synapse extracellular matrix component dystroglycan; staining for Vlgr1/Gpr98 (very large G protein-coupled receptor 1/G protein-coupled receptor 98) was mislocated. The Donating Investigator reports that this targeted mutation is a loss of function allele. This strain was generated from breeding (Stock No. 017760) mice to a germ line Cre recombinase expressing strain.
A targeting vector was used to insert a point mutation in exon 17 that results in an amino acid substitution of threonine for isoleucine at position 756, FRT site flanked PGKneo cassette into intron 17, as well as loxP sites into introns 13 and 17. The construct was electroporated into C57BL/6 derived Bruce4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts. The resulting chimeric animals were crossed to C57BL/6 mice. The mice were then crossed to Cre recombinase-expressing mice on the C57BL/6 genetic background to remove the selection cassette and exons 14 through 17.
|Allele Name||targeted mutation 1.1, Susanne tom Dieck|
|Allele Type||Targeted (Null/Knockout)|
|Gene Symbol and Name||Cacna1f, calcium channel, voltage-dependent, alpha 1F subunit|
|Strain of Origin||B6.Cg-Thy1a|
|Molecular Note||Germ line, cre mediated recombination was used to remove exons 14 through 17. Expression of a truncated protein was confirmed by western blot analysis on retinal extracts.|
|Mutations Made By|| |
When maintaining a live colony, homozygous females may be bred to hemizygous males(mutation is X-linked).
When using the B6.Cg-Cacna1ftm1.1Sdie/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #017761 in your Materials and Methods section.