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Fiziol Zh (1994) ; 60(3): 75-9, 2014.
Article in Ukrainian | MEDLINE | ID: mdl-25097934

ABSTRACT

We investigated the effect of proteases, widely used for neuron isolation in electrophysiological studies, on the amplitude and kinetic characteristics of persistent sodium current (I(NaP)) in hippocampal CA1 pyramidal neurons. Properties of I(NaP) were studied on neurons isolated by mechanical treatment (control group) and by mechanical and enzymatic treatment using pronase E (from Streptomyces griseus) or protease type XXIII (from Aspergillus oryzae). We show that in neurons isolated with pronase E kinetic of activation and density of I(NaP) was unaltered. Enzymatic treatment with protease type XXIII did not alter I(NaP) activation but result in significant decrease in I(NaP) density. Our data indicates that enzymatic treatment using pronase E for neuron isolation is preferable for investigation of I(NaP).


Subject(s)
Bacterial Proteins/pharmacology , Fungal Proteins/pharmacology , Neurons/drug effects , Peptide Hydrolases/pharmacology , Pronase/pharmacology , Sodium Channels/metabolism , Animals , CA1 Region, Hippocampal/cytology , CA1 Region, Hippocampal/drug effects , CA1 Region, Hippocampal/metabolism , Cell Separation/methods , Ion Transport , Membrane Potentials/drug effects , Membrane Potentials/physiology , Neurons/cytology , Neurons/metabolism , Patch-Clamp Techniques , Primary Cell Culture , Pronase/metabolism , Proteolysis , Rats , Sodium/metabolism
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