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 behavior (circadian rhythm, locomotor function, baseline and paroxysmal dyskinesia).

Transgenic Mouse Lines

Global BK channel knockout (Slo–/–, Kcnma1–/–)
Available from Jax Labs (Stock #035902 B6.129(FVB)-Kcnma1tm1Rwa/J)
Citation: JBC 279:36746-36752 (2004)
PCR genotyping protocol & strain info
Transnetyx genotyping (Probes: Kcnma1 WT and Neomycin)
MTA

KCNMA1 channelopathy mouse models (CRISPR gene-edited)

  • Available from our lab
  • Citation:  eLife (2022)
  • Transnetyx genotyping info
  • MTA
  • GOF:  Kcnma1N999S and Kcnma1D434G 
  • LOF:  Kcnma1H444Q

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

Conditional BK channel knockout, expressing tdTomato (Kcnma1fl)
Available from Jax Labs (Stock #035901  B6.Cg-Kcnma1tm1.1Alme/J)
Citation: Physiological Reports 3(11): e12612 (2015).  Genbank Accession # 
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 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)

HUMAN BK CHANNEL cDNA sequences

BKVYR (MG279689) was used in the following publications:

Characterization of new human KCNMA1 Loss-of-Function Mutations. Biophysical Journal 118(3)114A (2020).

Comparative Gain-of-Function Effects of KCNMA1-N999S Mutation on Human BK Channel Properties. J Neurophysiology 123:560 (2020).

Effects of single nucleotide polymorphisms in human KCNMA1 on BK current properties.  Frontiers in Molecular Neuroscience. 12:285(2019).

BKQEERL (MG279688) used in: Effects of single nucleotide polymorphisms in human KCNMA1 on BK current properties.  Frontiers in Molecular Neuroscience. 12:285(2019).

MOUSE BK CHANNEL cDNA 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’.  J 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)

Others 

MOUSE

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