These Syt1 targeted mice exhibit altered Ca2+ binding and are useful in studies of synaptic neurotransmission.
Dr. Thomas C. Sudhof, Stanford University School of Medicine
Mice homozygous for this targeted point mutation are viable and fertile and do not display any gross physical or behavioral abnormalities. No structural changes are introduced to the mutant protein and no major changes in protein levels are detected. Intrinsic Ca2+ binding and Ca2+ dependent phospholipid binding by the isolated C2A domain are severely reduced. The apparent Ca2+ affinity of the double C2 domain fragment is not altered, but tightness of the Ca2+ phospholipid/double C2 domain complex affinity is decreased. Unlike the D232N mutation (Stock No. 006386), increased synaptic depression is not found during repetitive stimulation. The mutation does not induce any major changes in synaptic transmission. This mutant mouse strain represents a model that may be useful in studies of synaptic neurotransmitter release.
A targeting vector was used to create a D238N substitution in the exon encoding residues 214-269 and insert a floxed neomycin resistance gene into the flanking intron. The construct was electroporated into 129P2/OlaHsd-derived E14.1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts. Chimeric animals were crossed with C57BL/6. The strain has been minimally backcrossed to C57BL/6 by the donating laboratory
|Allele Name||targeted mutation 6, Thomas C Sudhof|
|Allele Type||Targeted (Inserted expressed sequence)|
|Gene Symbol and Name||Syt1, synaptotagmin I|
|Promoter||Syt1, synaptotagmin I, mouse, laboratory|
|Strain of Origin||Not Specified|
|Molecular Note||An aspartate to asparagine missense mutation (D238N) was introduced into the C2A domain via homologous recombination. The mutation was made in the C2A domain. Western blot of brain homogenates from homozygous mutant animals verified the presence of protein expression.|
|Mutations Made By|| |
Dr. Thomas Sudhof, Stanford University School of Medicine
Homozygotes are viable and fertile. When maintaining a live colony, homozygous mice may be bred together.
When using the D238N mouse strain in a publication, please cite the originating article(s) and include JAX stock #006387 in your Materials and Methods section.