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|  Oregon Health and Science University |
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| Oregon Health and Science University |
Small-conductance calcium-activated potassium channels (SK channels) are gated solely by intracellular Ca2+ ions and are fundamental regulators of neuronal excitability. Our laboratory cloned the SK channel family and currently focuses on two main areas.
First, we are investigating the physiological roles of SK channels in hippocampus.
Second, the laboratory is testing the hypothesis that a given subtype of SK channel can serve multiple roles in the same neuron by differential subcellular localization and interactions with distinct sets of microdomain partner proteins, forming an array of Ca2+ signaling complexes.
Member:
Bond, Chris
"The LightCycler 480 Real-Time PCR System is a fully integrated multiwell-plate based real-time PCR platform for highly accurate qualitative and quantitative detection of nucleic acids. Building on the benefits of Roche's capillary-based LightCycler® Systems, it goes one step further in offering enhanced throughput, compatibility with automation equipment and maximum flexibility regarding hard- and software.
Providing novel ways to combine speed and accuracy without compromises, the LightCycler® 480 Real-Time PCR System meets the needs of a broad range of applications in research fields such as gene expression studies, discovery and analysis of genetic variation or array data validation."
"SK2-delta mice harbor a null allele of the Kcnn2 (potassium intermediate/small conductance calcium-activated channel, subfamily N, member 2; also called SK2) gene, and may be useful in studying the role of small-conductance calcium-activated potassium (SK) channels in after-hyperpolarization and action potentials of neuronal, inner ear (cochlea), and urinary bladder tissues."
"A loxP site was inserted 40 nucleotides 5' of the initiation methionine codon in exon 3 and a floxed neomycin resistance gene as well as an EGFP gene were inserted in the intron between exons 5 and 6. The EGFP insert was non functional. Cre mediated recombination removed the neo cassette leaving the loxP sites in exon 3 and intron 5 intact."
"The SK2T mutant allele results in approximately ten-fold overexpression of the Kcnn2 (potassium intermediate/small conductance calcium-activated channel, subfamily N, member 2; also called SK2) gene product prior to administration of tetracycline (or its analog doxycycline (dox))."
"The SK3T mutant allele has a tetracycline-based genetic switch inserted into the 5' UTR of the Kcnn3 (potassium intermediate/small conductance calcium-activated channel, subfamily N, member 3; also called SK3) locus, just upstream of the translation initiation site. This genetic switch harbors both the tetracycline-controlled transactivator protein (tTA) as well as the tetracycline operator (tetO; also called tetracycline-responsive element (TRE) or tet-operator); allowing transcription of the downstream Kcnn3 locus to be blocked by administration of tetracycline (or its analog doxycycline (dox)). Homozygotes exhibit three-fold overexpression of SK3 before, and no SK3 expression during, doxycycline administration."
"These floxed-SK3 mutant mice possess loxP sites flanking the translation initiation codon, coding sequences of exon 1, and a portion of intron1 of the potassium intermediate/small conductance calcium-activated channel, subfamily N, member 3 (Kcnn3) gene. SK3 is expressed in the soma and dendrites of dopaminergic neurons in the substantia nigra and influences their action potential frequency. Defects in dopamine (DA) releasing neurons have been sighted in pathologies such as schizophrenia and Parkinson's disease. SK3 is also expressed in smooth muscle of the bladder and uterus, and endothelia of the vasculature where the channels participate in blood pressure regulation. Mice that are homozygous for this allele are viable and fertile. When bred to mice that express tissue-specific Cre recombinase, resulting offspring will have exon 1 deleted in cre-expressing tissues."
Protocol for genotyping B6.129S4(Cg)-Kcnn2tm2Jpad/J mice.
This assay will NOT distinguish hemizygous from homozygous transgenic animals.
PCR protocol for genotyping Kcnn1tm1.2Jpad transgenic mice.
PCR protocol for genotyping Kcnn2tm1.1Jpad transgenic mice.
Protocol for genotyping Kcnn3tm1Jpad mice.
Genotyping protocol for Kcnn3tm2.1Jpad mouse strains.