MilliporeSigma
  • Home
  • Search Results
  • Knockout of Slo2.2 enhances itch, abolishes KNa current, and increases action potential firing frequency in DRG neurons.

Knockout of Slo2.2 enhances itch, abolishes KNa current, and increases action potential firing frequency in DRG neurons.

eLife (2015-11-13)
Pedro L Martinez-Espinosa, Jianping Wu, Chengtao Yang, Vivian Gonzalez-Perez, Huifang Zhou, Hongwu Liang, Xiao-Ming Xia, Christopher J Lingle
ABSTRACT

Two mammalian genes, Kcnt1 and Kcnt2, encode pore-forming subunits of Na(+)-dependent K(+) (KNa) channels. Progress in understanding KNa channels has been hampered by the absence of specific tools and methods for rigorous KNa identification in native cells. Here, we report the genetic disruption of both Kcnt1 and Kcnt2, confirm the loss of Slo2.2 and Slo2.1 protein, respectively, in KO animals, and define tissues enriched in Slo2 expression. Noting the prevalence of Slo2.2 in dorsal root ganglion, we find that KO of Slo2.2, but not Slo2.1, results in enhanced itch and pain responses. In dissociated small diameter DRG neurons, KO of Slo2.2, but not Slo2.1, abolishes KNa current. Utilizing isolectin B4+ neurons, the absence of KNa current results in an increase in action potential (AP) firing and a decrease in AP threshold. Activation of KNa acts as a brake to initiation of the first depolarization-elicited AP with no discernible effect on afterhyperpolarizations.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Ala-Gln, ≥98% (HPLC)
Sigma-Aldrich
Ala-Gln, BioReagent, suitable for cell culture, suitable for insect cell culture
Sigma-Aldrich
L-Cysteine, from non-animal source, BioReagent, suitable for cell culture, ≥98%
Sigma-Aldrich
L-Cysteine, ≥97%, FG
Sigma-Aldrich
L-Cysteine, 97%
Sigma-Aldrich
Ala-Gln, 200 mM, solution, sterile-filtered, Biotechnology Performance Certified
Sigma-Aldrich
Sodium dodecyl sulfate solution, BioUltra, for molecular biology, 20% in H2O
SAFC
L-Cysteine
Sigma-Aldrich
L-Cysteine, BioUltra, ≥98.5% (RT)
Sigma-Aldrich
Sodium dodecyl sulfate solution, BioUltra, for molecular biology, 10% in H2O
Sigma-Aldrich
Potassium hydroxide, anhydrous, ≥99.95% trace metals basis
Sigma-Aldrich
Potassium hydroxide, BioXtra, ≥85% KOH basis
Sigma-Aldrich
Potassium hydroxide, pellets, reag. Ph. Eur., ≥85%
Sigma-Aldrich
Potassium hydroxide, ACS reagent, ≥85%, pellets
Sigma-Aldrich
Potassium hydroxide solution, 45 wt. % in H2O
Sigma-Aldrich
Potassium hydroxide, reagent grade, 90%, flakes
Sigma-Aldrich
Potassium hydroxide, semiconductor grade, pellets, 99.99% trace metals basis (Purity excludes sodium content.)
Sigma-Aldrich
Potassium hydroxide, puriss. p.a., ≥86% (T), pellets
Sigma-Aldrich
Potassium hydroxide, free-flowing, Redi-Dri, ACS reagent, ≥85%, pellets
Sigma-Aldrich
Potassium hydroxide, technical, ≥85%, powder
Sigma-Aldrich
Potassium hydroxide, puriss., meets analytical specification of Ph. Eur., BP, 85-100.5%, pellets
Supelco
Potassium hydroxide solution, volumetric, 8.0 M KOH (8.0N)
Sigma-Aldrich
Potassium hydroxide, ≥85% KOH basis, pellets, white
Sigma-Aldrich
Potassium hydroxide on alumina
Supelco
Acetone solution, certified reference material, 2000 μg/mL in methanol: water (9:1)
Sigma-Aldrich
L-Cysteine, produced by Wacker Chemie AG, Burghausen, Germany, ≥98.0%
Sigma-Aldrich
HEPES, anhydrous, free-flowing, Redi-Dri, ≥99.5%
SAFC
HEPES
Sigma-Aldrich
Acetone, ACS reagent, ≥99.5%
Sigma-Aldrich
Acetone, histological grade, ≥99.5%