title: Impaired cardiac contractility in mice lacking both the AE3 Cl-/HCO3- exchanger and the NKCC1 Na+-K+-2Cl- cotransporter: effects on Ca2+ handling and protein phosphatases.
abstract: To analyze the cardiac functions of AE3, we disrupted its gene (Slc4a3) in mice. Cl(-)/HCO3(-) exchange coupled with Na+-dependent acid extrusion can mediate pH-neutral Na+ uptake, potentially affecting Ca2+ handling via effects on Na+/Ca2+ exchange. AE3 null mice appeared normal, however, and AE3 ablation had no effect on ischemia-reperfusion injury in isolated hearts or cardiac performance in vivo. The NKCC1 Na+-K+-2Cl(-) cotransporter also mediates Na+ uptake, and loss of NKCC1 alone does not impair contractility. To further stress the AE3-deficient myocardium, we combined the AE3 and NKCC1 knock-outs. Double knock-outs had impaired contraction and relaxation both in vivo and in isolated ventricular myocytes. Ca2+ transients revealed an apparent increase in Ca2+ clearance in double null cells. This was unlikely to result from increased Ca2+ sequestration, since the ratio of phosphorylated phospholamban to total phospholamban was sharply reduced in all three mutant hearts. Instead, Na+/Ca2+ exchanger activity was found to be enhanced in double null cells. Systolic Ca2+ was unaltered, however, suggesting more direct effects on the contractile apparatus of double null myocytes. Expression of the catalytic subunit of protein phosphatase 1 was increased in all mutant hearts. There was also a dramatic reversal, between single null and double null hearts, in the carboxymethylation and localization to the myofibrillar fraction, of the catalytic subunit of protein phosphatase 2A, which corresponded to the loss of normal contractility in double null hearts. These data show that AE3 and NKCC1 affect Ca2+ handling, PLN regulation, and expression and localization of major cardiac phosphatases and that their combined loss impairs cardiac function.
authors.affiliation: Department of Molecular Genetics, Biochemistry, and Microbiology, University of Cincinnati College of Medicine, Cincinnati, Ohio 45267-0524, USA.
authors.forename: Vikram
authors.initials: V
authors.lastname: Prasad
authors.forename: Ilona
authors.initials: I
authors.lastname: Bodi
authors.forename: Jamie W
authors.initials: JW
authors.lastname: Meyer
authors.forename: Yigang
authors.initials: Y
authors.lastname: Wang
authors.forename: Muhammad
authors.initials: M
authors.lastname: Ashraf
authors.forename: Sandra J
authors.initials: SJ
authors.lastname: Engle
authors.forename: Thomas
authors.initials: T
authors.lastname: Doetschman
authors.forename: Karena
authors.initials: K
authors.lastname: Sisco
authors.forename: Michelle L
authors.initials: ML
authors.lastname: Nieman
authors.forename: Marian L
authors.initials: ML
authors.lastname: Miller
authors.forename: John N
authors.initials: JN
authors.lastname: Lorenz
authors.forename: Gary E
authors.initials: GE
authors.lastname: Shull
mesh_terms: D000818:Animals; D017920:Antiporters / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism*; D002319:Cardiovascular System / Q000378:metabolism; D005298:Fertility; D051379:Mice; D018345:Mice, Knockout; D009154:Mutation / Q000235:genetics; D009200:Myocardial Contraction; D010749:Phosphoprotein Phosphatases / Q000378:metabolism*; D010766:Phosphorylation; D015427:Reperfusion Injury / Q000235:genetics / Q000378:metabolism; D028021:Sodium-Potassium-Chloride Symporters / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D064506:Solute Carrier Family 12, Member 2; D015854:Up-Regulation
chemical_list: D017920:Antiporters; C578878:Slc12a2 protein, mouse; C062002:Slc4a3 protein, mouse; D028021:Sodium-Potassium-Chloride Symporters; D064506:Solute Carrier Family 12, Member 2; D010749:Phosphoprotein Phosphatases; D002118:Calcium
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