L-type Ca2+ current downregulation in chronic human atrial fibrillation is associated with increased activity of protein phosphatases.
BACKGROUND
Although downregulation of L-type Ca2+ current (I(Ca,L)) in chronic atrial fibrillation (AF) is an important determinant of electrical remodeling, the molecular mechanisms are not fully understood. Here, we tested whether reduced I(Ca,L) in AF is associated with alterations in phosphorylation-dependent channel regulation.


RESULTS
We used whole-cell voltage-clamp technique and biochemical assays to study regulation and expression of I(Ca,L) in myocytes and atrial tissue from 148 patients with sinus rhythm (SR) and chronic AF. Basal I(Ca,L) at +10 mV was smaller in AF than in SR (-3.8+/-0.3 pA/pF, n=138/37 [myocytes/patients] and -7.6+/-0.4 pA/pF, n=276/86, respectively; P<0.001), though protein levels of the pore-forming alpha1c and regulatory beta2a channel subunits were not different. In both groups, norepinephrine (0.01 to 10 micromol/L) increased I(Ca,L) with a similar maximum effect and comparable potency. Selective blockers of kinases revealed that basal I(Ca,L) was enhanced by Ca2+/calmodulin-dependent protein kinase II in SR but not in AF. Norepinephrine-activated I(Ca,L) was larger with protein kinase C block in SR only, suggesting decreased channel phosphorylation in AF. The type 1 and type 2A phosphatase inhibitor okadaic acid increased basal I(Ca,L) more effectively in AF than in SR, which was compatible with increased type 2A phosphatase but not type 1 phosphatase protein expression and higher phosphatase activity in AF.


CONCLUSIONS
In AF, increased protein phosphatase activity contributes to impaired basal I(Ca,L). We propose that protein phosphatases may be potential therapeutic targets for AF treatment.
Christ
T
T
Department of Pharmacology,Dresden University of Technology, Dresden, Germany.
Boknik
P
P
Wöhrl
S
S
Wettwer
E
E
Graf
E M
EM
Bosch
R F
RF
Knaut
M
M
Schmitz
W
W
Ravens
U
U
Dobrev
D
D
D000368:Aged; D001281:Atrial Fibrillation / Q000201:enzymology* / Q000503:physiopathology*; D020746:Calcium Channels, L-Type / Q000378:metabolism*; D002908:Chronic Disease; D015536:Down-Regulation; D004553:Electric Conductivity; D004789:Enzyme Activation; D005260:Female; D006801:Humans; D007545:Isoproterenol / Q000494:pharmacology; D008297:Male; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000201:enzymology / Q000502:physiology; D009638:Norepinephrine / Q000494:pharmacology; D018408:Patch-Clamp Techniques; D010749:Phosphoprotein Phosphatases / Q000378:metabolism*; D011494:Protein Kinases / Q000378:metabolism
D020746:Calcium Channels, L-Type; D011494:Protein Kinases; D010749:Phosphoprotein Phosphatases; D007545:Isoproterenol; D009638:Norepinephrine