Common components of patch-clamp internal recording solutions can significantly affect protein kinase A activity

• Published in: Brain research Vol. 828; no. 1; pp. 169 - 173
• Main Authors: Vargas, Gricelly, Yeh, Tsung-Yin J, Blumenthal, Donald K, Lucero, Mary T
• Format: Journal Article
• Language: English
• Published: London Elsevier B.V 15-05-1999; Amsterdam Elsevier; New York, NY
• Subjects: Animals; Aspartic Acid - pharmacology; Biological and medical sciences; Calcium - pharmacology; Cells, Cultured; Cesium - pharmacology; Chelating Agents - pharmacology; Chlorides - pharmacology; Cyclic AMP-Dependent Protein Kinases - antagonists & inhibitors; Cyclic AMP-Dependent Protein Kinases - metabolism; Egtazic Acid - pharmacology; Enzyme Activation - drug effects; Enzyme Activation - physiology; Enzyme Inhibitors - pharmacology; Fluorides - pharmacology; Fundamental and applied biological sciences. Psychology; General aspects. Models. Methods; Gluconates - pharmacology; Ion channel; Ion Channels - physiology; Isoquinolines - pharmacology; Methylamines - pharmacology; Olfactory Receptor Neurons - chemistry; Olfactory Receptor Neurons - cytology; Olfactory Receptor Neurons - enzymology; Patch-Clamp Techniques; Phosphates - pharmacology; Phosphorylation; Potassium Compounds - pharmacology; Rats; Second messenger; Second Messenger Systems - drug effects; Second Messenger Systems - physiology; Serum Albumin, Bovine - pharmacology; Sulfonamides; Sulfuric Acid Esters - pharmacology; Tetraethylammonium - pharmacology; Vertebrates: nervous system and sense organs; Whole-cell voltage-clamp; Phosphorylation; Ion channel; Second messenger; Whole-cell voltage-clamp; Enzymatic activity; Enzyme; Protein kinase; Transferases; Electrophysiology; Patch clamp method; Ionic channel
• ISSN: 0006-8993; 1872-6240
• DOI: 10.1016/S0006-8993(99)01306-2

Common components of whole-cell internal recording solutions were tested both in vitro and in patch-clamp experiments for their effects on the activity of cAMP-dependent protein kinase. Potassium fluoride (KF), 440 mM trimethylamine chloride and exclusion of bovine serum albumin (BSA) decreased the activity of the enzyme, while ethylene glycol-bis (β-aminoethyl ether)
N,
N,
N′,
N′-tetraacetic acid (EGTA) and the potassium salts of aspartate, gluconate, methylsulfate and monobasic phosphate increased its activity. Addition of KF to the internal solution produced a hyperpolarizing shift in the
V
1/2 of
I
h channel activation, consistent with the KF-induced reduction of protein kinase A activity. Therefore, consideration of the composition of internal solutions is warranted when studying channel physiology by patch-clamp techniques.