TY - JOUR
T1 - Hyperpolarization-activated cyclic nucleotide-gated channels in pancreatic β-cell
AU - El-Kholy, Wasim
AU - MacDonald, Patrick E.
AU - Fox, Jocelyn Manning
AU - Bhattacharjee, Alpana
AU - Xue, Tian
AU - Gao, Xiaodong
AU - Zhang, Yi
AU - Stieber, Juliane
AU - Li, Ronald A.
AU - Tsushima, Robert G.
AU - Wheeler, Michael B.
PY - 2007/3
Y1 - 2007/3
N2 - Hyperpolarization-activated cyclic nucleotide-modulated (HCN) channels mediate the pacemaker current (Ih or If) observed in electrically rhythmic cardiac and neuronal cells. Here we describe a hyperpolarization-activated time-dependent cationic current, β-I h, in pancreatic β-cells. Transcripts for HCN1-4 were detected by RT-PCR and quantitative PCR in rat islets and MIN6 mouse insulinoma cells. β-Ih in rat β-cells and MIN6 cells displayed biophysical and pharmacological properties similar to those of HCN currents in cardiac and neuronal cells. Stimulation of cAMP production with forskolin/3-isobutyl-1- methylxanthine (50 μM) or dibutyryl-cAMP (1 mM) caused a significant rightward shift in the midpoint activation potential of β-Ih, whereas expression of either specific small interfering (si)RNA against HCN2 (siHCN2b) or a dominant-negative HCN channel (HCN1-AAA) caused a near-complete inhibition of time-dependent β-Ih. However, expression of siHCN2b in MIN6 cells had no affect on glucose-stimulated insulin secretion under normal or cAMP-stimulated conditions. Blocking β-Ih in intact rat islets also did not affect membrane potential behavior at basal glucose concentrations. Taken together, our experiments provide the first evidence for functional expression of HCN channels in the pancreatic β-cell.
AB - Hyperpolarization-activated cyclic nucleotide-modulated (HCN) channels mediate the pacemaker current (Ih or If) observed in electrically rhythmic cardiac and neuronal cells. Here we describe a hyperpolarization-activated time-dependent cationic current, β-I h, in pancreatic β-cells. Transcripts for HCN1-4 were detected by RT-PCR and quantitative PCR in rat islets and MIN6 mouse insulinoma cells. β-Ih in rat β-cells and MIN6 cells displayed biophysical and pharmacological properties similar to those of HCN currents in cardiac and neuronal cells. Stimulation of cAMP production with forskolin/3-isobutyl-1- methylxanthine (50 μM) or dibutyryl-cAMP (1 mM) caused a significant rightward shift in the midpoint activation potential of β-Ih, whereas expression of either specific small interfering (si)RNA against HCN2 (siHCN2b) or a dominant-negative HCN channel (HCN1-AAA) caused a near-complete inhibition of time-dependent β-Ih. However, expression of siHCN2b in MIN6 cells had no affect on glucose-stimulated insulin secretion under normal or cAMP-stimulated conditions. Blocking β-Ih in intact rat islets also did not affect membrane potential behavior at basal glucose concentrations. Taken together, our experiments provide the first evidence for functional expression of HCN channels in the pancreatic β-cell.
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U2 - 10.1210/me.2006-0258
DO - 10.1210/me.2006-0258
M3 - Article
C2 - 17158221
AN - SCOPUS:33847227276
SN - 0888-8809
VL - 21
SP - 753
EP - 764
JO - Molecular Endocrinology
JF - Molecular Endocrinology
IS - 3
ER -