Differential expression of two-pore domain potassium channels in rat cerebellar granule neurons

• Published in: Biochemical and biophysical research communications Vol. 453; no. 4; pp. 754 - 760
• Main Authors: Burgos, Paulina, Zúñiga, Rafael, Domínguez, Pedro, Delgado-López, Fernando, Plant, Leigh D., Zúñiga, Leandro
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 31-10-2014
• Subjects: Animals; Cells, Cultured; Cerebellar granule neurons (CGNs); Cerebellum - cytology; Cerebellum - metabolism; Gene Expression Regulation - physiology; Ion Channel Gating - physiology; K2P channels; Neurons - metabolism; Porosity; Potassium Channels - chemistry; Potassium Channels - classification; Potassium Channels - metabolism; Potassium Channels, Tandem Pore Domain - chemistry; Potassium Channels, Tandem Pore Domain - metabolism; Protein Structure, Tertiary; Rats; Rats, Sprague-Dawley; Two-pore domain potassium channels; K2P channels; Cerebellar granule neurons (CGNs); Two-pore domain potassium channels
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2014.10.012

•We examine the accumulation pattern of K2P channels transcripts in CGN.•K2P channels expressed in CGN were confirmed by Western blot and immunofluorescence.•K2P1, K2P3, K2P9 and K2P18 represent the majority component of IKSO current in CGN.
Two pore domain potassium (K2P) channels are mostly present in the central nervous system (CNS) where they play important roles in modulating neuronal excitability. K2P channels give rise to background K+ currents (IKSO) a key component in setting and maintaining the resting membrane potential in excitable cells. Here, we studied the expression and relative abundances of K2P channels in cerebellar granule neurons (CGNs), combining molecular biology, electrophysiology and immunologic techniques. The CGN IKSO was very sensitive to external pH, as previously reported. Quantitative determination of mRNA expression level demonstrated the existence of an accumulation pattern of transcripts in CGN that encode K2P9>K2P1>K2P3>K2P18>K2P2=K2P10>K2P4>K2P5 subunits. The presence of the major K2P subunits expressed was then confirmed by Western blot and immunofluorescence analysis, demonstrating robust expression of K2P1 (TWIK-1), K2P3 (TASK-1), K2P9 (TASK-3) and K2P18 (TRESK) channel protein. Based, on these results, it is concluded that K2P1, -3, -9 and -18 subunits represent the majority component of IKSO current in CGN.