Meredith Lab

Techniques

Molecular biology: generation of transgenic mice, RT-PCR, cloning, site-directed mutagenesis, transcript and protein expression, immunohistochemistry and in situ hybridization, microarray analysis of gene expression.

Electrophysiology: macroscopic currents, action potentials, single and multi-unit extracellular recordings, planar multi-electrode arrays, whole-cell recording, patch-clamp, acute brain slices, dissociated primary neuronal cultures, and organotypic cultures.

Systems physiology: telemetry (ECG, EEG, pressure, temperature), bladder cystometry, isometric tension recordings from smooth muscle, cardiovascular function, and circadian behavioral rhythms.

Transgenic Mouse Lines

BK channel knockout, global (Slo–/–, Kcnma1–/–)
Citation: JBC 279:36746-36752 (2004)
PCR genotyping protocol & strain info
Transnetyx genotyping (Probes: Kcnma1 WT and Neomycin)
MTA

BK channel gain-of-function, Per1-driven expression (Tg-BKR207Q)
Citation: PNAS 109(46):18997-9002 (2012).   Genbank Accession #JX429072.
Transnetyx genotyping (Probes: Kcnma1-2 Tg)
MTA

BK channel knockout, conditional, expressing tdTomato (Kcnma1fl)
Citation: Physiological Reports 3(11): e12612 (2015).  Genbank Accession # (coming
soon).
Transnetyx genotyping (Probes: Kcnma1-3 WT and tdRFP)
MTA

Beta2 subunit knockout, conditional, (Kcnmb2–/–), made by Chris Lingle (Wash U)
Citation: Nature Communications Mar 4;7:10837 (2016).
Original Citation for generation of mouse line: J Gen Physiol 144(4):275-95 (2014).
Available from us, Chris Lingle, or Jax Labs (Stock #028416).
Transnetyx genotyping (Probes: Kcnmb2-1 WT and Kcnmb2-1 EX)

Genbank Links for BK Variants (Organized by Publication)

MOUSE SEQUENCES

BK channels regulate sinoatrial node firing rate and cardiac pacing in vivo.  AJP- Heart 307(9):H1327-38 (2014).

– BKQEERL (KF530043), BKVYR (KF530042

Phosphorylation of a constitutive serine inhibits BK channel variants containing the alternate exon ‘SRKR’.  Journal of General Physiology 142 (6):585-598 (2013).

– BKSRKR (JX462785); Mutations: BKSRKR-S642A, , BKSRKR-R640Q/R645Q
– BKSRKR with YFP tag (JX462786)
– mbr5:YFP (KF530038)
– C2:YFP (KF530044)
– C3:YFP (KF530036)
– C4:YFP, also called BKVYR (KF530042)
– BK0:YFP (KF530040); Mutations: BK0-S642D

Mis-expression of the BK K+ channel disrupts suprachiasmatic nucleus circuit rhythmicity and alters clock-controlled behavior. AJP- Cell Physiology 304(4):C299-C311 (2013).

Genetic activation of BK currents in vivo generates bi-directional effects on neuronal excitability.  PNAS 109(46):18997-9002 (2012).
Tg-BKR207Q (JX429072)

Orphans (To be Published Soon!)

MOUSE

– C5c:YFP (KF530037)
– Aorta1 (Ao1:YFP) (KF530039)

HUMAN

– hBKQEERL
– hBKSRKR and BK0