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ELECTROPHYSIOLOGICAL STUDIES OF NEPHILA CLAVATA VENOM AND TOXIN INFLUENCE ON HIPPOCAMPAL NEURONAL...
ISSN 1996-4536 (print) • ISSN 2311-0783 (on-line) • Біологічні Студії / Studia Biologica • 2020 • Том 14/№3 • С. 29–48
Biol. Stud. 2020: 14(3); 29–48 • DOI: https://doi.org/10.30970/sbi.1403.626
www.http://publications.lnu.edu.ua/journals/index.php/biology
© 2020 O. M. Klyuchko; Published by the Ivan Franko National University of Lviv on behalf of Біологічні Cтудії / Studia Biologica. This is an Open
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UDC: 547.993.02:595.443.8-114.5.008
ELECTROPHYSIOLOGICAL STUDIES OF NEPHILA CLAVATA
VENOM AND TOXIN INFLUENCE ON HIPPOCAMPAL NEURONAL MEMBRANES
AFTER PRETREATMENT WITH ASPERGILLUS ORYZAE PROTEASES
O. M. Klyuchko
National Aviation University, 1, Lubomyr Huzar Ave., Kyiv 03058, Ukraine
e-mail: kelenaXX@nau.edu.ua
Klyuchko O.M. Electrophysiological studies of Nephila clavata venom and toxin infuence on
hippocampal neuronal membranes after pretreatment with Aspergillus oryzae proteases. Studia
Biologica, 2020: 14(3); 29–48 • DOI: https://doi.org/10.30970/sbi.1403.626
Background. The use of Arthropodae toxins for electrophysiological experiments is
very important, but experimental data in this regard are scarce. The aim of this study was
to obtain experimental data about the infuence of Nephila clavata spider venom and its
main active component – toxin JSTX-3 on the glutamate channel-receptor complex.
Methods. Kainate was used as agonist of glutamate channel-receptor complex
because it initiated non-inactivated transmembrane electric currents in rat hippocampal
membranes in electrophysiological experiments were used for the study. All chemicals
were applied to perfused hippocampal pyramidal neuronal membranes using ’concen-
tration-clamp’ technique and voltage-clamp recording.
Results and discussion. The studied substances – integral venom and toxin
JSTX-3 – demonstrated the properties of glutamate channel-receptor complex antago-
nists. The amplitudes of electric transmembrane currents activated by glutamate, kaina-
te, and quisqualate decreased (sometimes to zero) after the application of glutamate
channel-receptor complex antagonists to the rat hippocampal membrane under the
voltage-clamp conditions. The kinetics of activation and desensitization (in case of glu-
tamate and quisqualate) of transmembrane electric currents were not affected by these
antagonists.
The effects of Nephila clavata integral venom were studied the in concentrations of
10
-8
–10
-4
units/µL, the effects of JSTX-3 – in the concentrations of 10
-6
–10
-5
mol/L. Integral
venom did not block the studied currents completely, but it reduced their amplitudes to
a certain level. Integral venom blocked glutamate-activated currents up to 36±15 % of the
initial values, kainate-activated – up to 34±16 %. In contrast, JSTX-3 almost completely
blocked ion currents activated by these agonists at the holding potential of -100 mV: