Biomedical subjects

E M Volkov

Publications and source records attributed to E M Volkov.

At least 19 recordsLinked to original sources

PubMed2006

Effect of GABAergic and adrenergic agents on activity of Na+/K+ pump and Cl(-)-cotransport in somatic muscle cells of earthworm Lumbricus Terrestris.

GABA, baclofen, epinephrine, and norepinephrine hyperpolarized the membrane of earthworm somatic cells. This effect was prevented by furosemide, removal of Cl- from the medium, or activation of Na+/K+ pump by 3-fold increase external potassium concentration. It was hypothesized that GABA, baclofen, epinephrine, and norepinephrine stimulate Na+/K+ transport via specific receptor inputs, but their effect on resting potential can be realized only under conditions of working Cl- symport.

Adrenergic Agents↗

PubMed2005

Mechanisms of hyperpolarizing effect of GABA on resting potential of the Lumbricus terrestris muscular wall somatic cells.

GABA, baclofen, isoguvacine increase, and cis-4-aminocrotonic acid does not modify resting membrane potential of muscle cells. Bicuculline, phaclofen, N-ethylmaleimide, chlorpromazine, verapamil, and removal of Ca2+ from bathing solution abolished the effect of baclofen, while U73122 and D609 were ineffective in this respect. The authors conclude that the Lumbricus terrestris muscle cells contain GABAergic structures similar to a- and b-receptors. Activation of GABA receptors induced Cl- inward current and Ca2+ entry with subsequent activation of calmodulin-like proteins, which causes membrane hyperpolarization by increasing the effect of "pumping potential" on resting membrane potential.

Animals↗

PubMed2005

Effects of cholinergic receptor agonists and antagonists on miniature stimulatory postsynaptic ionic currents in somatic muscle cells of lumbricus terrestris.

Miniature stimulating postsynaptic currents of Lumbricus terrestris somatic muscle cells were recorded. Atropine, d-tubocurarin, alpha-bungarotoxin, carbacholine, and proserin did not modify the amplitude and temporal parameters of miniature stimulatory postsynaptic currents, while carbacholine and nicotine depolarized the muscle membrane. Presumably, Lumbricus terrestris muscle cells contain acetylcholine-sensitive channel-receptor complexes not belonging to classical nicotinic or muscarinic acetylcholine receptors.

Animals↗

PubMed2003

Effects of some transmitters on resting membrane potential of somatic cell in Lumbricus terrestris muscle wall.

Serotonin, glutamate, glycine, ATP, and muscarine had no effect on resting membrane potential of muscle cell in earthworm Lumbricus terrestris. Nicotine depolarizes and GABA hyperpolarizes the muscle membrane. Removal of K(+), Cl(-) and addition of ouabaine and strychnine to the solution abolished the effect of GABA. The authors conclude that the Lumbricus terrestris myocyte membrane contains nicotine receptors and GABAergic receptors sensitive to strychnine. Stimulation of these receptors activates sarcolemmal ionic pumps and causes membrane hyperpolarization.

Animals↗

PubMed2003

Two populations of miniature excitatory synaptic ionic currents in somatic muscle cells of Lumbricus terrestris earthworm body wall.

Three types of miniature excitatory synaptic currents were recorded in the same synaptic region of earthworm muscle cells: monoexponential (tau=1.2 msec) and biexponential (tau(1)=1.2 and tau(2)=8.0 msec). It was hypothesized that earthworm muscle cells contain at least two populations of acetylcholine-sensitive ionic channels, which do not belong to classical nicotinic and muscarinic acetylcholine receptors.

Animals↗

PubMed2003

Chloride cotransport in the membrane of earthworm body wall muscles.

The resting membrane potential (V(m)) of isolated somatic longitudinal muscles of the earthworm Lumbricus terrestris was studied by glass microelectrodes. The inhibition of chloride permeability by low pH did not affect V(m) of the muscle fibers in isolated somatic longitudinal muscles of the earthworm Lumbricus terrestris which was -48.7 mV (inside negative) at pH 7.3 and -49.1 at pH 5.6. On the other hand, bathing the muscles in Cl(-) and Na(+)-free solutions, or application of the chloride transporter inhibitor furosemide and Na(+)-K(+)-ATPase inhibitor ouabain depolarized the V(m) by 3-5 mV. The effects of a Cl(-) -free solution and ouabain were not additive. This demonstrates relatively small contribution of equilibrium potential for Cl(-) to the resting membrane potential and electrogenic effect of Na(+)K(+)-ATPase which is dependent on the supply of Na(+)(i) ions by furosemide-sensitive and Cl(-)(e)- and Na(+)(e)-dependent electroneutral transport (most probably Na(+)K(+)Cl(-) cotransport).

Animals↗

PubMed2002

Calcium mechanism of norepinephrine activation of ionic pump in somatic cells of lumbricus terrestris earthworm muscle wall.

Blockade of norepinephrine-induced hyperpolarization in earthworm muscle cells was observed in a calcium-free medium, after substitution of Ca(2+)with Mn(2+), and in the presence of verapamil or chlorpromazine to the incubation saline. Changes in Ca(2+)concentration in the saline and addition of caffeine had no effect on the resting potential of muscle cells. It was hypothesized that signal transduction from norepinephrine-activated membrane adrenoceptors to the ionic pump in earthworm muscle cells depends on influx of extracellular Ca(2+)and subsequent involvement of Ca(2+)-accepting proteins similar to calmodulin in vertebrates.

Adrenergic alpha-Agonists↗

PubMed2002

Effect of ionic medium on carbacholine-induced membrane depolarization in lumbricus terrestris somatic muscle cells.

Carbacholine depolarizes Lumbricus terrestris myocyte membrane. Addition of verapamil, tetrodotoxin, removal of K(+), Cl(-), and Ca(2+), and replacement of Ca(2+)with Mn(2+)in the bathing solution did not prevent the carbacholine-induced decrease in resting potential, but in a sodium-free medium carbacholine was ineffective. It was hypothesized that depolarization of the sarcolemma resulting from activation of cholinomimetic-sensitive channel-receptor complex of the Lumbricus terrestris somatic muscle cells is primarily determined by sodium permeability of the membrane.

Animals↗

PubMed2001

New chemically reactive dsDNAs containing single internucleotide monophosphoryldithio links: reactivity of 5'-mercapto-oligodeoxyribonucleotides.

Novel modified DNA duplexes with single bridging 5'-SS-monophosphoryldithio links [-OP(=O)-O(-)-SS-CH(2)-] were synthesized by autoligation of an oligonucleotide 3'-phosphorothioate and a 5'-mercapto-oligonucleotide previously converted to a 2-pyridyldisulfide adduct. Monophosphoryldisulfide link formation is not a stringent template-dependent process under the conditions used and does not require strong binding of the reactive oligomers to the complementary strand. The modified internucleotide linkage, resembling the natural phosphodiester bond in size and charge density, is stable in water, easily undergoes thiol-disulfide exchange and can be specifically cleaved by the action of reducing reagents. DNA molecules containing an internal -OP(=O)-O(-)-SS-CH(2)- bridge are stable to spontaneous exchange of disulfide-linked fragments (recombination) even in the single-stranded state and are promising reagents for autocrosslinking with cysteine-containing proteins. The chemical and supramolecular properties of oligonucleotides with 5'-sulfhydryl groups were further characterized. We have shown that under the conditions of chemical ligation the 5'-SH group of the oligonucleotide has a higher reactivity towards N-hydroxybenzotriazole-activated phosphate in an adjacent oligonucleotide than does the OH group. This autoligation, unlike disulfide bond formation, proceeds only in the presence of template oligonucleotide, necessary to provide the activated phosphate in close proximity to the SH-, OH- or phosphate function.

Chromatography, High Pressure Liquid↗

PubMed2001

Effects of norepinephrine and epinephrine on resting membrane potential in body wall muscle cells of Lumbricus terrestris earthworm.

Norepinephrine and to a lesser extent epinephrine increased the resting membrane potential of earthworm body wall muscle cells. Ouabain, phentolamine, propranolol, and replacement of Ca(2+) with Mg(2+) in the incubation medium abolished this effect. External 3'5'-cAMP in high concentration, dibutyryl cAMP, and dibutyryl cGMP did not induced hyperpolarization of muscle cell membranes. It was concluded that norepinephrine and epinephrine increased the resting membrane potential in earthworm body wall muscle cells via activation of Na(+), K(+) pump. This process involves Ca(2+) and sarcolemmal adrenergic structures, similar to alpha-adrenoceptors in vertebrate, but not the cyclic nucleotide systems.

Adrenergic alpha-Agonists↗

PubMed2001

Effect of cholinergic agonists on resting membrane potential of earthworm body wall muscle cells.

Carbachol in a concentration of 5x10(-8) mol/liter does not hyperpolarize, and in a concentration of 5x10(-6) mol/liter depolarizes the membrane of somatic muscle cell in earthworm. d-tubocurarine, alpha-bungarotoxin, atropine, and hexamethonium added to the incubation medium did not abolish the carbachol-induced decrease in resting membrane potential. Each of these drugs alone had no effect on resting membrane potential in muscle cells. Presumably, the acetylcholine-sensitive receptor-channel membrane complex in earthworm muscle cell differs from acetylcholine receptor in skeletal muscle fibers and peripheral neurons of vertebrates.

Acetylcholine↗

PubMed2001

[Influence of sodium pump and Na(+), K(+), CL(-)-cotransport on the resting membrane potential of somatic muscle cells of the earthworm Lumbricus terrestris].

Tetrodotoxin and acidic pH do not change the resting membrane potential (RMP), whereas Na+ or Cl- free solutions or ouabain and furosemide equally depolarize the membrane of the earthworm somatic muscle cells. The findings of the RMP depending on extracellular K+ concentration corroborate theoretical model by Goldman-Hodgkin-Katz only in Na(+)-free medium or in presence of ouabain. The data suggest that the RMP is the sum of potassium and chlorine diffusion potentials as well as of the potential produced by electrogenic component of Na+ pump and, probably, by furosemide-sensitive Na+,K+Cl- co-transport.

Animals↗

PubMed2000

Targeting of single-stranded DNA and RNA containing adjacent pyrimidine and purine tracts by triple helix formation with circular and clamp oligonucleotides.

The aim of this work was to construct an anti-messenger targeted to the pim-1 oncogene transcript, based on circular or clamp oligodeoxyribonucleotides. The formation of bimolecular triplexes by clamp or circular oligonucleotides was investigated using single-stranded targets of both DNA (5'-CCCTCCTTTGAAGAA-3') and RNA type (5'-CCCUCCUUUGAAGAA-3'). The third, 'Hoogsteen' strand of the triplex was represented by G,T-rich sequences. The secondary structures of the complexes were determined by thermal denaturation, circular dichroism and gel mobility shift experiments and shown to depend on the nature of the target strand. With DNA as target, the sequence of a clamp (or circular) oligonucleotide that formed the triple helix was 3'-GGGAGGAAACTTCTTTT-TTGTTGTTT-TT-GGTGGG-5', where the first TT dinucleotide (in italics) is a linker and the second TT (bold) represents the bridge through which the 'Hoogsteen' strand switches from one strand of the Watson-Crick duplex to the other, once the duplex is formed by the corresponding portion of the anti-messenger (underlined). The portion of the 'Hoogsteen' sequence of the triplex between the two TT dinucleotides binds to the 3' extremity of the target strand and runs parallel to it. The portion situated at the 5' end of the oligonucleotide switches to the purine tract of the complementary strand of the duplex and is antiparallel to it. In contrast, with RNA as target, for a branched clamp oligonucleotide that formed a triple helix over its entire length (5'-TTCTTCAAAGGAGGG-3' 3'-GGGTGGTTT-T-GTTGTT-5') the portion of the 'Hoogsteen' sequence that bound to the 3' extremity of the target strand had to be antiparallel to it.

Circular Dichroism↗

PubMed2000

[Effect of the medium composition on the resting membrane potential in the earthworm longitudinal somatic muscle fibers].

Norepinephrine or increased extracellular K+ hyperpolarize the membrane of the earthworm somatic muscle fibre, whereas removal of Cl- from external solution or a hypotonic solution depolarize the membrane. The dependence of the membrane resting potential on the extracellular K+ is quite characteristic against the background of ouabain action. A preliminary membrane depolarisation by ouabain eliminates the above effects on the membrane resting potential. The data obtained suggest that the ouabain-sensitive active ion pump directly contributes to the membrane resting potential value. This hypothesis is discussed with respect to existence of active Cl- transport combined with Na+, K(+)-pump which presumably takes part in the intracellular osmotic pressure regulation in the earthworm somatic muscle.

Animals↗

PubMed2000

[A rapid method for testing the activity of the repair enzyme uracil-DNA-glycosylase].

A rapid and effective method of testing of a repair enzyme, uracil-DNA-glycosylase, was proposed. As a substrate, a deoxyuridine-containing 5'-32P-labeled deoxyoligonucleotide covalently attached to a polystyrene support (Tenta Gel S-NH2) was used. The ammonia cleavage of the apyrimidine site formed in the enzymic reaction followed by the transition of the labeled oligonucleotide fragment from the solid phase into solution allowed the detection of the enzymic activity.

Base Sequence↗

PubMed2000

Participation of electrogenic Na+-K+-ATPase in the membrane potential of earthworm body wall muscles.

The effect of Na+-K+-ATPase inhibitor ouabain on the resting membrane potential (Vm) was studied by glass microelectrodes in isolated somatic longitudinal muscles of the earthworm Lumbricus terrestris and compared with frog sartorius muscle. In earthworm muscle, Vm was -49 mV (inside negative) in a reference external solution with 4 mmol/l K+. The electrogenic participation of Na+-K+-ATPase was absent in solutions with very low concentrations of 0.01 mmol/l K+, higher in 4 and 8 mmol/l K+ (4-5 mV) and maximal (13 mV) in solutions containing 12 mmol/l K+ where Vm was -46 mV in the absence and -33 mV in the presence of 1 x 10(4) M ouabain. The electrogenic participation of Na+-K+-ATPase was much smaller in m. sartorius of the frog Rana temporaria bathed in 8 and 12 mmol/l K+. The results indicate that the Na+-K+-ATPase is an important electrogenic factor in earthworm longitudinal muscle fibres and that its contribution to Vm depends directly on the concentration of K+ in the bathing solution.

Animals↗

PubMed1999

Oligonucleotide-peptide conjugates as potential antisense agents.

Oligonucleotide-peptide conjugates have several applications, including their potential use as improved antisense agents for interfering with the RNA function within cells. In order to provide robust and generally applicable conjugation chemistry, we developed a novel approach of fragment coupling of pre-synthesized peptides to the 2'-position of a selected nucleotide within an otherwise protected oligonucleotide chain attached to a solid support.

Adenosine↗

PubMed1998

Role of DNA definite structural elements in interaction with repair enzyme uracil-DNA glycosylase.

Interaction of different oligodeoxyribonucleotides (oligos), their analogs and oligonucleopeptide with uracil-DNA glycosylase (UDG) from human placenta was investigated. It is shown that there is no considerable contribution of heterocyclic bases of DNA to UDG-substrate binding but the UDG interaction with some DNA phosphate groups is necessary for enzyme-substrate recognition. However the phosphate group adjacent to single dU from the 3'-end in oligo is not involved into the electrostatic contact with UDG. It is found that UDG has the high affinity to its reaction product. An oligonucleotide containing a single 2'-deoxy-2'-aminouridine is a non-hydrolyzable substrate analog for UDG.

DNA↗