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ISSN 2522-9028 (Print)
ISSN 2522-9036 (Online)
DOI: https://doi.org/10.15407/fz

Fiziologichnyi Zhurnal

is a scientific journal issued by the

Bogomoletz Institute of Physiology
National Academy of Sciences of Ukraine

Editor-in-chief: V.F. Sagach

The journal was founded in 1955 as
1955 – 1977 "Fiziolohichnyi zhurnal" (ISSN 0015 – 3311)
1978 – 1993 "Fiziologicheskii zhurnal" (ISSN 0201 – 8489)
1994 – 2016 "Fiziolohichnyi zhurnal" (ISSN 0201 – 8489)
2017 – "Fiziolohichnyi zhurnal" (ISSN 2522-9028)

Fiziol. Zh. 2018; 64(2): 26-33


OPTIMIZATION OF THE METHOD OF NUCLEI ISOLATION FOR ELECTROPHYSIOLOGICAL STUDIES OF ION CHANNELS IN THE NUCLEAR MEMBRANE OF THE RAT CARDIOMYOCYTES

O.A. Kotyk, A.B. Kotliarova, S.M. Marchenko

    O.O. Bogomoletz Institute of Physiology National Academy of Sciences of Ukraine, Kyiv, Ukraine
DOI: https://doi.org/10.15407/fz64.02.026

Abstract

The conditions for the nuclei isolation from cardiomyocytes for electrophysiological studies of ion channels in the native nuclear membrane were optimized. The structural and functional intactness of isolated nuclei is crucial for investigation of the biophysical properties of the transport systems in the native nuclear membrane. The quality of the obtained sample was evaluated by ion currents recording through the nuclear membrane in nucleus-attached or excised patch configuration at voltage-clamp mode of patch-clamp technique. In our study the solutions with different compositions were tested and the quality of the samples obtained using different methods of myocardium homogenization were compared. We found that the activity of ion channels in the nuclear membrane were most often registered when myocardium was homogenized in solution with the next composition (mmol/l): sucrose – 300, KCl – 60, HEPES – 10 (pH 7,2) with protease inhibitors cocktail, DL-dithiothreitol (1 mmol/l) and glycerol, using glass homogenizer (Dounce).

Keywords: nuclear envelope; cardiomyocytes; nuclear fraction isolation; native membrane; patch-clamp; ion channels

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