Paired pulse facilitation of summated intracellular synaptic potentials in single retinotectal fibers in the frog

Facilitation of the second of two consecutive test EEG quanta (the summated monosynaptic potentials of the synapses of one axon arborizing in layer F of the frog tectum) was investigated in the normal and under conditions of raised extracellular Ca²⁺ and Mg²⁺ concentration. Intensification of paired-pulse facilitation (×1.4–2.4) was observed at the shortest interstimulus intervals (of 2.5–5 msec). The distribution of maximum levels for facilitation of EEG quanta was bimodal at levels 1.95 and 1.65, on the basis of which two groups were identified, one potentiating EEG quanta more than the other. The time course of paired-pulse facilitation of both groups of EEG quanta can be broken down into two exponential components with time constants of 5–6, 140–150 and 6–8, 60–70 msec respectively. Bimodal distribution of maximum paired-pulse levels in the normal, together with findings from experiments involving raised Ca²⁺ and Mg²⁺ concentrations would indicate that facilitation of frog retinotectal synapses is dependent on the quantal release of transmitter; it may thus be postulated that this release reaches near-saturation point in the normal. It is suggested that two types of axonal terminal arborizations whose synapses differ in the quantal content of transmitter release are present in layer F of the frog tectum. These axonal arbors could well originate from different class 3 and 5 retinal detectors.Z. Yanushkevichyus Institute of Physiology and Pathology of the Cardiovascular System of the Kaunas Medical Institute. Translated from Neirofiziologiya, Vol. 18, No. 1, pp. 45–55, January–February, 1986.     

Action of thiamine on auditory evoked potentials in the guinea pig

A technique is proposed for quantifying the effects of physiologically active substances at the periphery of the auditory analyzer. It was found that applying 1×10^–11 to 1×10^–3 M thiamine to the membrane of guinea pig cochlear round window (fenestra rotunda) produces a rise in the amplitude and a reduction in the latency of the N₁ and N₂ components of auditory nerve action potentials, waves I and II of brainstem auditory evoked potentials occurring in response to an acoustic stimulus. It is suggested that this effect is produced by facilitated synaptic transmission at synapses between hair cells and spiral ganglia neurons under the action of thiamine penetrating into the cochlea.A. V. Palladin Institute of Biochemistry, Academy of Sciences of the Ukrainian SSR, Kiev. A. I. Kolomiichenko Research Institute of Otolaryngology, Ministry of Public Health of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 18, No. 5, pp. 654–660, September–October, 1986.     

Effects of ethimizol on frog neuromuscular junction

The effects were studied of ethimizol, a substance activating memory processes, on features of synaptic transmission during experiments on frog cutaneous pectoris muscle. It was found that the presynaptic action of ethimizol consists of raising the frequency of miniature potentials, when used at a concentration of 0.5–10 mM, and modulating quantal content of synaptic transmission due to changes in binomial quantal release parameters p and n when 0.5–2 mM ethimizol was used. This substance facilitated transmission at synapses with a low initial level of transmitter release. This substance facilitated transmission at synapses with a low initial level of transmitter release. Ethimizol was also found to have a postsynaptic action, consisting of reducing amplitude at a concentration of 5–10 mM and prolonging synaptic currents and potentials when concentrations of 0.5–10 mM were used. The latter effect produced a considerable increase in the time integral of endplate potentials. The postsynaptic action of ethimizol is perhaps seen in its effects on features of postsynaptic ionic channels. The effects of ethimizol are discussed with a view to how it may act within the central nervous system as a nonspecific modulator.A. A. Zhdanov Leningrad State University. Translated from Neirofiziologiya, Vol. 17, No. 6, pp. 757–763, November–December, 1985.     

Effect of dalargin,a synthetic leu-enkephalin analogue on synaptic transmission in the Lorenzini ampullae of rays

Effects of dalargin, a synthetic leu-enkephalin analogue and its antagonist naloxone on synaptic transmission in afferent synapses of ray electroreceptors were investigated using an isolated preparation of Lorenzini ampullae from Black sea rays. It was shown that dalargin (10^–6–10^–10 mole liter) both decreased background activity and evoked activity of an afferent fiber in a dose-dependent manner. Naloxone (10^–5 mole/liter) also inhibited afferent impulsation and completely blocked responses of the Lorenzini ampullae to dalargin application. L-glutamate-induced excitatory responses were reduced in the presence of dalargin. It is suggested that the modulatory action of dalargin on glutamatergic synaptic transmission in the Lorenzini ampullae is exerted via specific opiate receptors.Translated from Neirofiziologiya, Vol. 25, No. 1, pp. 18–21, January–February, 1993.     

Comparison of synaptosomal and glial uptake of pipecolic acid and GABA in rat brain

The active uptake of [³H]pipecolic acid increased with incubation time and its uptake at 3 min was half of that at 20 min. [¹⁴C]GABA uptake rose earlier, and its uptake at 3 min was almost 80% of that at 20 min. On the other hand, a ratio (pellet/medium) of [³H]pipecolic acid uptake into glial cell-enriched fractions, was much less (0.4–0.6) than that of [¹⁴C]GABA (25.8–74.1). GABA, 10^–4 M, and pipecolic acid, 10^–4 M, produced a significant inhibition of [³H]pipecolic acid uptake into P₂ fractions. Pipecolic acid, 10^–4 M, significantly reduced the synaptosomal and glial uptake of [¹⁴C]GABA. GABA, 10^–4 M, affected neither spontaneous nor high K⁺-induced release of [³H]pipecolic acid from brain slices. It is suggested that pipecolic acid is involved in either synaptic transmission or in its modulation at GABA synapses in the central nervous system.     

Modulation of synaptic events by heavy metals in the central nervous system of mollusks

Summary 1. The effects of heavy metals (Pb²⁺, Hg²⁺, and Zn²⁺) on synaptic transmission in the identified neural network ofHelix pomatia L. andLymnaea stagnalis L. (Gastropoda, Mollusca) were studied, with investigation of effects on inputs and outputs as wells as on interneuronal connections.2. The sensory input running from the cardiorenal system to the central nervous system and the synaptic connections between central neurons were affected by heavy metals.3. Lead and mercury (10^–5–10^–3 M) eliminated first the inhibitory, then the excitatory inputs running from the heart to central neurons. At the onset of action lead increased the amplitude of the excitatory postsynaptic potentials, but blockade of sensory information transfer occurred after 10–20 min of treatment.4. The monosynaptic connections between identified interneurons were inhibited by lead and mercury but not by zinc. Motoneurons were found to be less sensitive to heavy metal treatment than interneurons or sensory pathways.5. The treatment with Pb²⁺ and Hg²⁺ often elicited pacemaker and bursting-type firing in central neurons, accompanied by disconnection of synaptic pathways, manifested by insensitivity to sensory synaptic influences.6. Zn²⁺ treatment also sometimes induced pacemaker activity and burst firing but did not cause disconnection of the synaptic transmission between interneurons.7. A network analysis of heavy metal effects can be a useful tool in understanding the connection between their cellular and their behavioral modulatory influences.     

Wavelet Transform-Based De-Noising for Two-Photon Imaging of Synaptic Ca2+ Transients

Postsynaptic Ca²⁺ transients triggered by neurotransmission at excitatory synapses are a key signaling step for the induction of synaptic plasticity and are typically recorded in tissue slices using two-photon fluorescence imaging with Ca²⁺-sensitive dyes. The signals generated are small with very low peak signal/noise ratios (pSNRs) that make detailed analysis problematic. Here, we implement a wavelet-based de-noising algorithm (PURE-LET) to enhance signal/noise ratio for Ca²⁺ fluorescence transients evoked by single synaptic events under physiological conditions. Using simulated Ca²⁺ transients with defined noise levels, we analyzed the ability of the PURE-LET algorithm to retrieve the underlying signal. Fitting single Ca²⁺ transients with an exponential rise and decay model revealed a distortion of τ_rise but improved accuracy and reliability of τ_decay and peak amplitude after PURE-LET de-noising compared to raw signals. The PURE-LET de-noising algorithm also provided a ∼30-dB gain in pSNR compared to ∼16-dB pSNR gain after an optimized binomial filter. The higher pSNR provided by PURE-LET de-noising increased discrimination accuracy between successes and failures of synaptic transmission as measured by the occurrence of synaptic Ca²⁺ transients by ∼20% relative to an optimized binomial filter. Furthermore, in comparison to binomial filter, no optimization of PURE-LET de-noising was required for reducing arbitrary bias. In conclusion, the de-noising of fluorescent Ca²⁺ transients using PURE-LET enhances detection and characterization of Ca²⁺ responses at central excitatory synapses.     

Synaptic apparatus of the feline primary auditory cortex (area Al)

Electron microscope research was conducted on the synaptic apparatus of the feline primary auditory cortex (Al). A total of 2096 profiles of axonal terminals (AT) were found over a total area of 8230 µm² of ultrathin slices at different layers of this cortical layer — an average of 255 profiles per 1000 µm² of the surface area on these slices. The AT profiles occupied about 8.9% of the surface of these cross-sections. It was found that 52% of the AT containing synaptic vesicles formed asymmetrical or symmetrical synaptic contacts (83.9% and 16.1% respectively) and that AT had no contacts which could be considered synaptic junctions on 48% of slices. It was also observed that 45.3% of the AT forming contacts synapsed on spines, 48.5% on dendrites, and 6.2% on neuronal somata. Finally, 95.4% and 4.6% of axo-spinal synapses contained rounded and flattened vesicles respectively; equivalent figures for axodendritic synapses were 79.4% and 20.6% respectively and 19.8 and 80.2% for axosomatic synapses. Calculations revealed an average of 322.8 × 10⁶ AT over 1 mm³ of cat auditory cortex. Organizational aspects of synaptic apparatus at different layers of area A1 were ascertained.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 22, No. 4, pp. 533–543, July–August, 1990.     

Investigation of heterosynaptic interactions in hippocampal areas CA1, and CA3 in vitro

Heterosynaptic interactions between synapses located at a considerable distance from the cell body (perforant path) and lying close to the body of the neuron (synapses of Schaffer's collaterals and axons of the dentate fascia) on guinea pig hippocampal neurons were investigatedin vitro. It was shown by the paired stimulus method that, using stimulation of subthreshold intensity for action potential generation, spatiotemporal summation takes place in both pairs of synaptic systems. If above-threshold stimulation was used, afferents lying close to the cell body suppressed responses evoked by stimulation of distant afferents for a longer time (up to 20 msec in area CA₁ and up to 300 msec in area CA₃) than during the opposite combination of stimuli (up to 3–8 msec). After tetanization of the dentate fascia depression of responses of area CA₃ neurons to stimulation of the perforant path was observed for 2–30 min. In the remaining cases, no significant prolonged heterosynaptic posttetanic changes were observed. The possible mechanisms of these interactions are discussed.Institute of Biophysics, Academy of Sciences of the USSR, Pushchino-on-Oka. Translated from Neirofiziologiya, Vol. 11, No. 6, pp. 524–532, November–December, 1979.     

Layer by layer analysis of synaptic organization in the optic tectum of the prog

The distribution of axo-axonal and axo-dendritic synapses, nerve endings, and bodies of neurons by depth in the optic tectum ofRana temporaria L. was investigated under normal conditions and 6–9, 60, and 134 days after removal of the contralateral eye. Counting was carried out on long oriented sections examined in the electron microscope. In outer plexiform layer 9 the density of synapses was greatest near the surface of the tectum and decreased in the direction away from it; no inner sublayers with differing density of synapses could be distinguished. In the outer zone of layer 9 (to a depth of about 30 ) many axo-axonal synpases were discovered. Endings of myelinated optic fibers of large diameter ("dark" terminal degeneration) were widely distributed in the same layer. The density of axo-dendritic synapses in deep plexiform layer 5 was similar to that in layer 9. Many nerve endings containing granular vesicles as well as pale synaptic vesicles were found in layer 5 and neighboring zones.A. N. Severtsov Institute of Evolutionary Morphology and Ecology of Animals, Academy of Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 11, No. 2, pp. 130–136, March–April, 1979.     

[125I]calmodulin binding to synaptic plasma membrane from rat brain: Kinetic and arrhenius analysis

Binding of [¹²⁵I]calmodulin was characterized in highly purified synaptic plasma membrane (SPM) prepared from rat brain. By Scatchard analysis, the Ca²⁺-dependent membrane binding of [¹²⁵I]calmodulin was found to have a B_max of 284 pmol/mg protein and an apparent affinity with a K_d of 131 nM. Kinetic analysis indicates that at 37°C, the dissociation of [¹²⁵I]calmodulinmembrane complexes follows first-order reaction and consists of two components: a dissociation constant (k) of 3.7×10^–1 min^–1 and a half-time (t_1/2) of 1.8 min for the fast component, and a k of 4.8×10^–2 min^–1 and a t_1/2 of 14.5 min for the slow component. At 0°C, substantial dissociation still occurred, with a k of 4.5×10^–2 min^–1 and a t_1/2 of 15.3 min for the fast component, and a k of 5.5×10^–3 min^–1 and a t_1/2 of 125.5 min for the slow component. These data on binding affinity and dissociation kinetics are consistent with the notion that SPM can readily and rapidly associated and dissociate calmodulin. In Arrhenius analysis of temperature effects, [¹²⁵I]calmodulin binding to SPM exhibits a biphasic function, with the transition temperature (T_d) estimated to be 23.8°C, suggesting that binding is influenced by lipid phase transition of the membrane. The binding of [¹²⁵I]calmodulin to the synaptic membrane was found to be increased by corticosterone (10^–7–10^–6 M), a steroid hormone, and decreased by ethanol (50–200 mM), a centrally acting drug. Our data on the characteristics of calmodulin binding to the SPM provide groundwork for future studies on physiological and pharmacological regulation of calmodulin translocation to and from the plasma membrane in synaptic terminals.Abbreviations used CaM calmodulin - SPM synaptic plasma membrane - ATPase adenosine triphosphatase - Tris tris(hydroxymethyl)aminomethane - EGTA ethylene-bis(oxyethylenenitrilo)tetraacetic acid - SDS sodium dodecyl sulfate - TFP trifluoperazine - K_d dissociation constant - B_max maximum binding - k first-order rate constant - t_1/2 half-time - T_d transition temperature     

Axo-axonal synapses in the frog tectum opticum

A quantitative electron-microscopic investigation of synaptic endings in large sections showed that about 50% of all axo-axonal synapses are located in the outer zone of the neuropil (layer 9) of the tectum opticum ofRana temporaria L. These synapses are more numerous in the rostral part of the tectum than the caudal. Hardly any axo-axonal synapses lie deeper than 50–60 µ Most axo-axonal synapses are located on axon endings of retinal ganglionic cells, for after degeneration of the optic nerve the number of these synapses is reduced by two-thirds. During ontogenetic differentiation and regeneration of the optic nerve axo-axonal synapses develop before axo-dendritic and their presynaptic processes have the normal structure and differ sharply from the bulbs of growth of the optic fibers. On this basis the central origin of most presynaptic processes forming these synapses is postulated. The results point to the possibility of presynaptic control over the effectiveness of action of the efferent axons, primarily optic, terminating in the outer zone of the frog tectum opticum.     

Structure of Excitatory Synapses and GABAA Receptor Localization at Inhibitory Synapses Are Regulated by Neuroplastin-65

Formation, maintenance, and activity of excitatory and inhibitory synapses are essential for neuronal network function. Cell adhesion molecules (CAMs) are crucially involved in these processes. The CAM neuroplastin-65 (Np65) highly expressed during periods of synapse formation and stabilization is present at the pre- and postsynaptic membranes. Np65 can translocate into synapses in response to electrical stimulation and it interacts with subtypes of GABA_A receptors in inhibitory synapses. Here, we report that in the murine hippocampus and in hippocampal primary culture, neurons of the CA1 region and the dentate gyrus (DG) express high Np65 levels, whereas expression in CA3 neurons is lower. In neuroplastin-deficient (Np^−/−) mice the number of excitatory synapses in CA1 and DG, but not CA3 regions is reduced. Notably this picture is mirrored in mature Np^−/− hippocampal cultures or in mature CA1 and DG wild-type (Np^+/+) neurons treated with a function-blocking recombinant Np65-Fc extracellular fragment. Although the number of GABAergic synapses was unchanged in Np^−/− neurons or in mature Np65-Fc-treated Np^+/+ neurons, the ratio of excitatory to inhibitory synapses was significantly lower in Np^−/− cultures. Furthermore, GABA_A receptor composition was altered at inhibitory synapses in Np^−/− neurons as the α1 to α2 GABA_A receptor subunit ratio was increased. Changes of excitatory and inhibitory synaptic function in Np^−/− neurons were confirmed evaluating the presynaptic release function and using patch clamp recording. These data demonstrate that Np65 is an important regulator of the number and function of synapses in the hippocampus.     

NMDA receptors in afferent synapses of frog semicircular canals

The effects of competitive (2-amino-phosphonovaleric acid) and noncompetitive (Mg²⁺, ketamine, kynurenic acid) antagonists of N-methyl-D-aspartate (NMDA) receptors on synaptic transmission were studied in afferent synapses of the frog semicircular canals. All of these antagonists reduced the rate of background activity in the nerve of posterior semicircular canal by 30–50%, which confirms the presence of glutamate NMDA receptors in the hair cell synapses in the frog semicircular canals.Neurofiziologiya/Neurophysiology, Vol. 25, No. 3, pp. 168–169, May–June, 1993.     

Conducting pathways of the rabbit lumbar sympathetic ganglia

Conducting pathways of ganglia from the lumbar portion (L₃–L₅) of the sympathetic trunk in rabbits were studied by recording action potentials from nerves of the ganglia evoked by stimulation of other nerves of these ganglia, and by intracellular recording from single neurons. Besides the well-known system of descending preganglionic fibers, which enter the trunk through white rami communicantes and, as they pass through the ganglia, form synapses on ganglionic neurons, some preganglionic fibers were shown to enter the sympathetic chain through gray rami communicantes and to run in both ascending and descending directions, forming synaptic connections with neurons of the lumbar ganglia.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 16, No. 2, pp. 247–254, March–April, 1984.     

Neurotransmitter receptor and time dependence of the synaptic plasticity disrupting actions of Alzheimer's disease Aβ in vivo

Many endogenous factors influence the time course and extent of the detrimental effects of amyloid β-protein (Aβ) on synaptic function. Here, we assessed the impact of varying endogenous glutamatergic and cholinergic transmission by pharmacological means on the disruption of plasticity at hippocampal CA3-to-CA1 synapses in the anaesthetized rat. NMDA receptors (NMDARs) are considered critical in mediating Aβ-induced inhibition of long-term potentiation (LTP). However, intracerebroventricular injection of Aβ_1–42 inhibited not only NMDAR-dependent LTP but also voltage-activated Ca²⁺-dependent LTP induced by strong conditioning stimulation during NMDAR blockade. On the other hand, another form of NMDAR-independent synaptic plasticity, endogenous acetylcholine-induced muscarinic receptor-dependent long-term enhancement, was not hindered by Aβ_1–42. Interestingly, augmenting endogenous acetylcholine activation of nicotinic receptors prior to the injection of Aβ_1–42 prevented the inhibition of NMDAR-dependent LTP, whereas the same intervention when introduced after the infusion of Aβ was ineffective. We also examined the duration of action of Aβ, including water soluble Aβ from Alzheimer''s disease (AD) brain. Remarkably, the inhibition of LTP induction caused by a single injection of sodium dodecyl sulfate-stable Aβ dimer-containing AD brain extract persisted for at least a week. These findings highlight the need to increase our understanding of non-NMDAR mechanisms and of developing novel means of overcoming, rather than just preventing, the deleterious synaptic actions of Aβ.     

Transmission Efficacy and Plasticity in Glutamatergic Synapses Formed by Excitatory Interneurons of the Substantia Gelatinosa in the Rat Spinal Cord

Background

Substantia gelatinosa (SG, lamina II) is a spinal cord region where most unmyelinated primary afferents terminate and the central nociceptive processing begins. The glutamatergic excitatory interneurons (EINs) form the majority of the SG neuron population, but little is known about the mechanisms of signal processing in their synapses.

Methodology

To describe the functional organization and properties of excitatory synapses formed by SG EINs, we did non-invasive recordings from 183 pairs of monosynaptically connected neurons. An intact presynaptic SG EIN was specifically stimulated through the cell-attached pipette while the evoked EPSCs/EPSPs were recorded through perforated-patch from a postsynaptic neuron (laminae I-III).

Principal Findings

We found that the axon of an SG EIN forms multiple functional synapses on the dendrites of a postsynaptic neuron. In many cases, EPSPs evoked by stimulating an SG EIN were sufficient to elicit spikes in a postsynaptic neuron. EPSCs were carried through both Ca²⁺-permeable (CP) and Ca²⁺-impermeable (CI) AMPA receptors (AMPARs) and showed diverse forms of functional plasticity. The synaptic efficacy could be enhanced through both activation of silent synapses and strengthening of already active synapses. We have also found that a high input resistance (R_IN, >0.5 GΩ) of the postsynaptic neuron is necessary for resolving distal dendritic EPSCs/EPSPs and correct estimation of their efficacy.

Conclusions/Significance

We conclude that the multiple synapses formed by an SG EIN on a postsynaptic neuron increase synaptic excitation and provide basis for diverse forms of plasticity. This functional organization can be important for sensory, i.e. nociceptive, processing in the spinal cord.     

Nerve cells and synaptic connections in the intestinal nerve of the snail,Helix pomatia L.

Summary The ultrastructure of nerve cells and the finestructural organization of synaptic contacts have been investigated in the intestinal nerve in the snail Helix pomatia. Three types of nerve cells, occurring singly or in groups, can be distinguished on the basis of the ultrastructure of their perikaryon and content of granules. The peripheral output of these nerve cells has been verified by retrograde CoCl₂ and NiCl₂ staining. Both axosomatic and axo-axonic specialized synaptic contacts occur in the intestinal nerve. Presynaptic elements of these synaptic contacts contain 100–120 nm granular vesicles or 120–200 nm neurosecretory-like granules. Following intracellular horseradish peroxidase (HRP) labelling of identified central neurons responsible for peripheral regulatory processes, several labelled axons running toward the periphery can be followed throughout the branches of the intestinal nerve. These labelled axon processes (either primary axon or small collaterals) form specialized synaptic contacts, inside the intestinal nerve, and are always in a postsynaptic position. The occurrence of peripheral axo-somatic and axo-axonic synapses provides a morphological basis for integrative processes taking place in the intestinal nerve (peripheral nervous system) of Helix pomatia.     

Kinetics of Na+, K+-ATPase Inhibition by a Rat Brain Endogenous Factor (II-E)

Previous work from this laboratory led to the isolation by gel filtration and anionic exchange HPLC of a rat brain fraction named II-E, which highly inhibits synaptosomal membrane Na⁺, K⁺-ATPase activity. In this study we evaluated the kinetics of such inhibition and found that inhibitory potency was independent of Na⁺(1.56–200 mM), K⁺(1.25–40 mM), or ATP (1–8 mM) concentration. Hanes-Woolf plots indicated that II-E decreases Vmax but does not alter K_Mvalue, and suggested uncompetitive inhibition for Na⁺, K⁺or ATP. However, II-E became a stimulator at 0.5 mM ATP concentration. It is postulated that this brain factor may modulate ionic transport at synapses, thus participating in central neurotransmission.     

Polymodal distribution of synaptic delays in the frog neuromuscular junction

Synaptic delay of single-quantum response with low mean quantal size (0.05–1) was measured during experiments on preparations of frog neuromuscular junctions using extracellular focal recording of presynaptic action potentials and endplate currents. It was found that distribution of these synaptic delays is of a polymodal nature and mean intermodal interval equaled 0.22±0.01 msec over 13 experiments. An increase in quantal size produced only a redistribution of mode weighting, while mean modal interval remained unchanged. A reduction in temperature induced an increase in the modal interval with the temperature coefficient Q₁₀=2.42±0.14 (n=15). The explanation is suggested that the process of quantal transmitter release is determined by interaction between the calcium-dependent mechanism for raising the likelihood of release on the one hand and the rhythmic operation of the system producing transmitter release on the other. The latter stage in the process depends on temperature, not intracellular Ca²⁺ concentration. The polymodal distribution of synaptic delay reflects the rhythmic operation of the transmitter release zone.I. M. Sechenov Institute of Evolutionary Physiology and Biochemistry, Academy of Sciences of the USSR, Leningrad. Translated from Neirofiziologiya, Vol. 18, No. 6, pp. 748–756, November–December, 1986.