The oscillatory responses of skate electroreceptors to small voltage stimuli

Tonic nerve activity in skate electroreceptors is thought to result from spontaneous activity of the lumenal membranes of the receptor cells which is modulated by applied stimuli. When physiological conditions are simulated in vitro, the receptor epithelium produces a current which flows inward across the lumenal surface. This epithelial current exhibits small spontaneous sinusoidal fluctuations about the mean that are associated with corresponding but delayed fluctuations in postsynaptic response. Small voltage stimuli produce damped oscillations in the epithelial current similar in time-course to the spontaneous fluctuations. For lumen-negative, excitatory stimuli, these responses are predominantly an increase over the mean inward current. For inhibitory stimuli they are predominantly a decrease. Increased inward current across the lumenal membranes of the receptor cells increases depolarization of the presynaptic membranes in the basal faces leading to increased release of transmitter and an excitatory postsynaptic response. Decreased inward current decreases depolarization of the presynaptic membranes leading to a reduction in transmitter release and an inhibitory postsynaptic response. Clear changes in postsynaptic response are detectable during stimuli as small as 5 microV with saturation occurring at +/- 400 microV. The evoked oscillations in epithelial current are damped and the postsynaptic responses decline during maintained stimuli with large off-responses occurring at stimulus termination. The initial peak of the off-response is similar to the response produced by onset of an oppositely directed stimulus. These observations substantiate the role of receptor cell excitability in the detection of small voltage changes.     

Processing of mechanosensory information from gustatory receptors on a hind leg of the locust

Gustatory receptors (basiconic sensilla) on the legs of the desert locust, Schistocerca gregaria, are innervated by chemosensory afferents and by a mechanosensory afferent. We show, for the first time, that these mechanosensory afferents form an elaborate detector system with the following properties: 1) they have low threshold displacement angles that decrease with increasing stimulus frequency in the range 0.05–1 Hz, 2) they respond phasically to deflections of the receptor shaft and adapt rapidly to repetitive stimulation, 3) they encode the velocity of the stimulus in their spike frequency and have velocity thresholds lower than 1°/s, and 4) they are directionally sensitive, so that stimuli moving proximally towards the coxa elicit the greatest response.The mechanosensory afferents, but not the chemosensory afferents, make apparently monosynaptic connections with spiking local interneurones in a population with somata at the ventral midline of the metathoracic ganglion. They evoke excitatory synaptic potentials that can sum to produce spikes in the spiking local interneurones. Stimulation of the single mechanosensory afferent of a gustatory receptor can also give rise to long lasting depolarizations, or to bursts of excitatory postsynaptic potentials in the interneurones that can persist for several seconds after the afferent spikes. These interneurones are part of the local circuitry involved in the production of local movements of a leg. The mechanosensory afferents from gustatory receptors must, therefore, be considered as part of the complex array of exteroceptors that provide mechanosensory information to these local circuits for use in adjusting, or controlling locomotion.     

Glutamate excitatory effects on ampullar receptors of the frog

Summary The action of glutamate on frog ampullar receptors was investigated to assess the potential role of this excitatory amino acid as an afferent transmitter in the hair cell system. Intracellular recordings from single afferent units in the isolated labyrinth revealed that glutamate and the glutamate receptor agonists, N-methyl-D-aspartic acid, quisqualic acid and kainic acid increase dose-dependently the frequency of the resting afferent discharge of EPSPs and spikes and produce long lasting depolarizations. After blocking synaptic transmission by using 5 mM Co²⁺, the same compounds elicited only depolarizations of amplitude comparable to those observed in normal saline. Quisqualic acid and kainic acid were much more potent than N-methyl-D-aspartic acid in increasing the frequency of afferent discharge and in causing axonal depolarizations. The depolarization caused by glutamate was reduced dose-dependently by the competitive non-NMDA receptor antagonist 6-cyano-7-nitroquinaxoline-2,3 dione and disappeared almost completely in Na⁺-free Ringer solution. These results are consistent with the hypothesis that glutamate is the afferent transmitter in vestibular organs and indicate that receptors mainly of the non-NMDA type are present not only at postsynaptic level but also in hair cells. Presynaptic glutamate receptors may function as autoreceptors controlling by a positive feed-back mechanism the release of the afferent transmitter.     

The organization of exteroceptive information from the uropod to ascending interneurones of the crayfish

The organization of exteroceptive inputs to identified ascending interneurones of the crayfish, Procambarus clarkii (Girard), has been analyzed by stimulation of hairs on the uropod and simultaneous intracellular recordings from ascending interneurones. The spikes of single afferent neurones which innervated hairs on the distal ventral surface of the exopodite were consistently followed by a depolarizing synaptic potential in many identified ascending interneurones with a constant and short central delay of 0.7–1.5 ms. The amplitude of the potentials depended on the membrane potential of the ascending interneurones. Each afferent neurone made divergent outputs onto several ascending interneurones and each ascending interneurone received convergent inputs from several afferent neurones. Certain ascending interneurones made inhibitory or excitatory connections with other ascending interneurones. These central interactions were always one-way, and the spikes from one ascending interneurone consistently evoked excitatory or inhibitory post-synaptic potentials in other interneurones which followed with a constant and short latency of 0.7–1.0 ms. The inhibitory postsynaptic potential was reversed by injection of steady hyperpolarizing current.Abbreviations EPSP excitatory post-synaptic potential - IPSP inhibitory post-synaptic potential     

Action of botulinum A toxin and tetanus toxin on synaptic transmission

Intracellular recordings of the spontaneous activity from mammalian spinal cord neurons in culture demonstrated different sensitivities of excitatory and inhibitory synaptic transmission for the action of tetanus toxin (Tetx) and botulinum toxin type A (Botx). The effects of Tetx and Botx on spontaneous and nerve-evoked transmitter release were compared under identical experimental conditions in experiments on in vitro poisoned mouse diaphragms. At 37 degrees C completely paralyzed endplates are characterized by a very low frequency of spontaneous miniature endplate potentials (m.e.p.p.s) and by a 100% failure to evoke endplate potentials (e.p.p.s) in response to single nerve stimuli. Striking differences in the action of both toxins have been observed when the very low transmitter release probabilities of paralyzed nerve-muscle preparations were increased by tetanic nerve stimulation and/or application of potent K+-channel blockers and/or by reduction of temperature to 25 degrees C. While Botx did not change the short latency between nerve impulse and postsynaptic response, Tetx produced a temporal dispersion of the quantal release suggesting that the toxins act at different sites in the chain of events that result in transmitter release. To find further evidence to support the different actions of the toxins the spontaneous transmitter release was studied in more detail. Tetx blocked preferentially the release of so-called large mode m.e.p.p.s without affecting the frequency of the small mode ones. In contrast, Botx strongly inhibited both the small and large mode m.e.p.p.s.(ABSTRACT TRUNCATED AT 250 WORDS)     

Maintained depolarization of synaptic terminals facilitates nerve-evoked transmitter release at a crayfish neuromuscular junction

Presynaptic and postsynaptic potentials were examined by intracellular recording at a crayfish neuromuscular junction. During normal synaptic transmission, the action potentials were recorded in the terminal region of the excitatory axon and postsynaptic responses were obtained in the muscle fibers. We found that it was possible to modify the synaptic transmission by applying depolarizing or hyperpolarizing currents through the presynaptic intracellular electrode. Typically, a 7-15 mV depolarization lasting longer than 50 msec leads to a large (500%) enhancement of transmitter release, even though the preterminal action potential is reduced in amplitude. Hyperpolarization increases the amplitude of the action potential, but slightly reduces the transmitter release. These results are different from those reported for other neuromuscular synapses and the squid giant synapse, but are similar in many respects to the results reported for several invertebrate central synapses. We conclude, first, that different synapses may have markedly different responses to conditioning by membrane polarization and, secondly, that maintained low-level depolarization may induce a potentiated state in the nerve terminal, perhaps brought about by slow entry of calcium.     

Can presynaptic depolarization release transmitter without calcium influx?

Recent experimental evidence suggesting that presynaptic depolarization can evoke transmitter release without calcium influx has been re-examined. The presynaptic terminal of the squid giant synapse can be depolarized by variable amounts while recording presynaptic calcium current under voltage clamp and postsynaptic responses. Small depolarizations open few calcium channels with large single channel currents. Large depolarizations approaching the calcium equilibrium potential open many channels with small single channel currents. When responses to small and large depolarizations eliciting similar total macroscopic calcium currents are compared, the large pulses evoke more transmitter release. This apparent voltage-dependence of transmitter release may be explained by the greater overlap of calcium concentration domains surrounding single open calcium channels when many closely apposed channels open at large depolarizations. This channel domain overlap leads to higher calcium concentrations at transmitter release sites and more release for large depolarizations than for small depolarizations which open few widely dispersed channels. At neuromuscular junctions, a subthreshold depolarizing pulse to motor nerve terminals may release over a thousand times as much transmitter if it follows a brief train of presynaptic action potentials than if it occurs in isolation. This huge synaptic facilitation has been taken as indicative of a direct effect of voltage which is manifest only when prior activity raises presynaptic resting calcium levels. This large facilitation is actually due to a post-tetanic supernormal excitability in motor nerve terminals, causing the previously subthreshold test pulse to become suprathreshold and elicit a presynaptic action potential. When motor nerve terminals are depolarized by two pulses, as the first pulse increases above a certain level it evokes more transmitter release but less facilitation of the response to the second pulse.(ABSTRACT TRUNCATED AT 250 WORDS)     

Chemically Mediated Transmission at a Giant Fiber Synapse in the Central Nervous System of a Vertebrate

The hatchetfish, Gasteropelecus, possesses large pectoral fin adductor muscles whose simultaneous contraction enables the fish to dart upwards at the approach of a predator. These muscles can be excited by either Mauthner fiber. In the medulla, each Mauthner fiber forms axo-axonic synapses on four "giant fibers," two on each side of the midline. Each pair of giant fibers innervates ipsilateral motoneurons controlling the pectoral fin adductor muscles. Mauthner fibers and giant fibers can be penetrated simultaneously by microelectrodes close to the synapses between them. Electrophysiological evidence indicates that transmission from Mauthner to giant fiber is chemically mediated. Under some conditions miniature postsynaptic potentials (PSP's) are observed, suggesting quantal release of transmitter. However, relatively high frequency stimulation reduces PSP amplitude below that of the miniature potentials, but causes no complete failures of PSP's. Thus quantum size is reduced or postsynaptic membrane is desensitized. Ramp currents in Mauthner fibers that rise too slowly to initiate spikes can evoke responses in giant fibers that appear to be asynchronous PSP's. Probably both spikes and ramp currents act on the same secretory mechanism. A single Mauthner fiber spike is followed by prolonged depression of transmission; also PSP amplitude is little affected by current pulses that markedly alter presynaptic spike height. These findings suggest that even a small spike releases most of an immediately available store of transmitter. If so, the probability of release by a single spike is high for any quantum of transmitter within this store.     

Inhibition and facilitation in parasympathetic ganglia of the urinary bladder

Neurons in vesical parasympathetic ganglia receive excitatory and inhibitory inputs from both divisions of the autonomic nervous system. Sacral parasympathetic pathways (cholinergic) provide the major excitatory input to these ganglia via activation of nicotinic receptors. Parasympathetic pathways also activate muscarinic inhibitory and excitatory receptors, which may exert a modulatory influence on transmission. Cholinergic transmission is relatively inefficient when preganglionic nerves are stimulated at low frequencies (< 1 Hz). However, excitatory postsynaptic potentials (EPSPs) and postganglionic firing markedly increase during repetitive stimulation at frequencies of 1-10 Hz. It is concluded that enhanced transmitter release accounts for the temporal facilitation and that vesical ganglia function as "high pass filters" that amplify the parasympathetic excitatory input to the detrusor muscle during micturition. Transmission in vesical ganglia is also sensitive to adrenergic inhibitory and facilitatory synaptic mechanisms elicited by efferent pathways in the hypogastric nerves. The effects of exogenous norepinephrine indicate that adrenergic inhibition is mediated by alpha receptors and reflects primarily a presynaptic depression of transmitter release although postsynaptic adrenergic hyperpolarizing and depolarizing effects have also been noted. Adrenergic facilitation is mediated by beta receptors as well as unidentified receptors. Norepinephrine also can inhibit or excite spontaneously active neurons in vesical ganglia. The existence of inhibitory and facilitatory synaptic mechanisms in vesical ganglia provides the basis for a complex ganglionic modulation of the central autonomic outflow to the bladder.     

Effects of aspartic acid on skate electroreceptors

The effects of L-aspartic acid (L-ASP) on spontaneous and evoked activity in afferent nerve fibers were investigated by perfusing the basal membrane of sea skate electroreceptors (the ampullae of Lorenzini) with this substance. It was found that perfusion with physiological saline containing L-ASP exerted a primarily excitatory effect on afferent activity (threshold concentration: 10^–7 M). When synaptic transmission was blocked by magnesium ions, activity was restored in the afferent fibers if L-ASP was added to the solution and spike activity persisted for longer; this would imply the presence of desensitizing processes in the postsynaptic receptors of the ampullae. Finding would lead to the conclusion that L-ASP and L-glutamate fulfill a set of criteria for likely neurotransmitters in the ampullae of Lorenzini.I. P. Pavlov Institute of Physiology, Academy of Sciences of the USSR, Leningrad. Translated from Neirofiziologiya, Vol. 19, No. 1, pp. 61–67, January–February, 1987.     

Structural and functional studies of the king salmon, Oncorhynchus tshawytscha, hemoglobins

Vertical starch-gel electrophoresis at pH 8.6 revealed extensive hemoglobin multiplicity with several distinct cathodal and anodal hemoglobin components. Anodal hemoglobin components are present throughout the life cycle of the king salmon. Additional cathodal components are found in the adult fish. Cathodal hemoglobin components exhibited a higher oxygen affinity (P50 = 10.2 mm at 13 degrees C, pH 7.3) than the anodal hemoglobin components (P50 = 21.8 mmHg at 13 degrees C). Oxygen binding of the anodal hemoglobins are sensitive to pH, temperature, organic phosphates (ATP and GTP), as well as, ionic strength; binding of oxygen to the cathodal hemoglobins is independent of pH and not affected by organic phosphates. Anodal hemoglobin components are less resistant to thermal denaturation over the pH 6.0 to 8.0 range. Isothermal urea denaturation of separated anodal and cathodal hemoglobin fractions of the king salmon indicate inherent differences in the stabilization energies of these hemoglobins. Autoxidation of these hemoglobins occurs around pH 7.0 and below, as well as, in the presence of increasing Cl- concentrations.     

Intracellular recordings from spinal neurons during 'swimming' in paralysed amphibian embryos

Intracellular microelectrode recordings have been made from probable motoneurons in the spinal cord of Xenopus laevis embryos during fictive 'swimming' in preparations paralysed with the neuromuscular blocking agent tubocurarine. These cells had resting potentials of -50 mV or more. During spontaneous or stimulus-evoked 'swimming' episodes: (a) the cells were tonically excited; the level of tonic synaptic excitation and the conductance increase underlying it were both inversely related to the 'swimming' cycle period; (b) the cells usually fired one spike per cycle in phase with the motor root burst on the same side; spikes did not overshoot zero and were evoked by phasic excitatory synaptic input on each cycle, superimposed on the tonic excitation; (c) in phase with motor root discharge on the opposite side of the body, the cells were hyperpolarized by a chloride-dependent inhibitory postsynaptic potential. The nature of synaptic potentials during 'swimming' was evaluated by means of intracellular current injections. The 'swimming' activity could be controlled by natural stimuli. The results provide clear evidence on the relation of tonic excitation to rhythmic locomotory pattern generation, and indirect evidence for reciprocal inhibitory coupling between antagonistic motor systems.     

Neural mechanism generating firing patterns in jaw motoneurons during the food-induced response in Aplysia kurodai

1. In each right and left buccal ganglia of Aplysia kurodai, we identified 4 premotor neurons impinging on the ipsilateral jaw-closing and -opening motoneurons. Three of them (MA1 neurons) had features of multifunctional neurons. Current-induced spikes in the MA1 neurons produced excitatory junction potentials (EJPs) in the buccal muscle fibers. In addition, tactile stimulation of the buccal muscle surface produced a train of spikes in the MA1 neurons without synaptic input. The other neuron (MA2) had only a premotor function. 2. The MA1 and MA2 neurons had similar synaptic effects on the jaw-closing and -opening motoneurons. Current-induced spikes in the premotor neurons gave rise to monosynaptic inhibitory postsynaptic potentials (IPSPs) in the ipsilateral jaw-closing motoneurons. Simultaneously, spikes in one of the MA1 neurons and the MA2 also gave rise to monosynaptic excitatory postsynaptic potentials (EPSPs) in the ipsilateral jaw-opening motoneuron. 3. The IPSPs and the EPSPs induced by spikes in the premotor neurons were reversibly blocked by d-tubocurarine and hexamethonium, respectively, suggesting that the MA1 and MA2 neurons are cholinergic. 4. When depolarizing and hyperpolarizing current pulses were passed into one premotor neuron, attenuated but similar potential changes were produced in another randomly selected premotor neuron in the same ganglion, suggesting that they are electronically coupled.     

Elementary polysynaptic potentials of hippocampal neurons

Synaptic connections of 26 pairs of hippocampal neurons were studied in nonanesthetized rabbits by spike-triggered averaging of intracellularly recorded activity. Synchronized activity was detected in 5 pairs and considered to represent common inputs to the neurons recorded. Hyperpolarizing or depolarizing potentials with 3--4 ms latency were revealed in 3 additional pairs. These potentials are considered to be individual postsynaptic potentials (PSPs) evoked in the target neuron by spikes of the adjacent (0.5--2 mm apart) neuron. A quantum analysis of the individual PSPs was performed. The mean quantum content (0.27--0.65) and quantum size (35--200 mkV) were found to be of the same order as those of the excitatory PSPs previously recorded after intracerebral stimulation. It is concluded that most hippocampal synapses are of low efficacy.     

Synaptic diversity and differentiation: Crustacean neuromuscular junctions

Crustacean motor neurons exhibit a wide range of synaptic responses. Tonically active neurons generally produce small excitatory postsynaptic potentials (EPSPs) at low impulse frequencies, and are able to release much more transmitter as the impulse frequency increases. Phasic neurons typically generate large EPSPs in their target cells, but have less capability for frequency facilitation, and undergo synaptic depression during maintained activity. These differences depend in part upon the neuron's ongoing levels of activity; phasic neurons acquire physiological and morphological features of tonic neurons when their activity level is altered. Molecules responsible for adaptation to activity can be sought in single identified phasic neurons with current techniques. The fact that both phasic and tonic neurons innervate the same target muscle fibers is evidence for presynaptic determination of synaptic properties, but there is also evidence for postsynaptic determination of specific properties of different endings of a single neuron. The occurrence of high- and low-output endings of the same tonic motor neurons on different muscle fibers suggests a target-specific influence on synaptic properties. Structural variation of synapses on individual terminal varicosities leads to the hypothesis that individual synapses have different probabilities for release of transmitter. We hypothesize that structurally complex synapses have a higher probability for release than the less complex synapses. This provides an explanation for the larger quantal contents of high-output terminals (where the proportion of complex synapses is higher), and also a mechanism for progressive recruitment of synapses during frequency facilitation.     

Electrophysiological investigation of the properties of the electroreceptors (ampullae of Lorenzini) in Black Sea rays

Responses of electroreceptors (ampullae of Lorenzini) in Black Sea rays to electrical stimuli were recorded in vivo as spike activity of single nerve fibers. Depending on their functional properties the fibers could be divided into silent, those with regular activity (10–15 spikes/sec) and those with grouped activity. Electrical stimuli evoked a tonic response with a varied degree of adaptation in the nerve fibers. The threshold currents were between 10^?10 and 10^?11 A/mm². The minimal latent period of the on-responses to pulses of current of maximal intensity was 15–40 msec, whereas that of the off-responses was 15–200 msec. The effect of intensity, duration, and polarity of the stimuli on the responses of the receptors and the adaptation of the electroreceptors during application of a steady current were investigated. The properties of the ampullae of Lorenzini were compared with those of other types of electroreceptors.     

Multivesicular release at climbing fiber-Purkinje cell synapses.

Synapses driven by action potentials are thought to release transmitter in an all-or-none fashion; either one synaptic vesicle undergoes exocytosis, or there is no release. We have estimated the glutamate concentration transient at climbing fiber synapses on Purkinje cells by measuring the inhibition of excitatory postsynaptic currents (EPSCs) produced by a low-affinity competitive antagonist of AMPA receptors, gamma-DGG. The results, together with simulations using a kinetic model of the AMPA receptor, suggest that the peak glutamate concentration at this synapse is dependent on release probability but is not affected by pooling of transmitter released from neighboring synapses. We propose that the mechanism responsible for the elevated glutamate concentration at this synapse is the simultaneous release of multiple vesicles per site.     

Nonclassical synaptic functions of transmitters

Nonclassical synaptic functions are considered in two groups, mainly by reference to the models provided by sympathetic ganglia. Slow postsynaptic potentials (PSPs) are compared with classical fast PSPs. Features include loose delivery of transmitter to receptor, very long synaptic delays and durations of PSPs, slow removal of transmitter or of its effects, integration of repetitive inputs, electrogenesis without large increases in ionic conductances. Neuromodulatory actions affect synaptic efficacy without direct excitatory or inhibitory responses to the transmitter. These include a) control of presynaptic release, and b) contingent postsynaptic actions. In b, a modulatory transmitter alters the efficacy of action by another transmitter. The alteration may persist long after exposure to the modulatory transmitter; in mammalian sympathetic ganglia, exposure to dopamine or to a conditioning train of preganglionic volleys induces a long-term enhancement of the muscarinic slow excitatory PSP. Or the alteration may be restricted mostly to the presence of a modulatory transmitter, with examples cited. Nonclassical synaptic functions may be providing revolutionary possibilities for dealing with slow and broadly distributed cerebral functions, manifested electrophysiologically and behaviorally, that have been difficult to analyze successfully in terms restricted to the fast and discretely localized classical synaptic functions.     

Calcium-activated conductance in skate electroreceptors: current clamp experiments

When current clamped, skate electroreceptor epithelium produces large action potentials in response to stimuli that depolarize the lumenal faces of the receptor cells. With increasing stimulus strength these action potentials become prolonged. When the peak voltage exceeds about 140 mV the repolarizing phase is blocked until the end of the stimulus. Perfusion experiments show that the rising phase of the action potential results from an increase in calcium permeability in the lumenal membranes. Perfusion of the lumen with cobalt or with a zero calcium solution containing EGTA blocks the action potential. Perfusion of the lumen with a solution containing 10 mM Ca and 20 mM EGTA initially slows the repolarizing process at all voltages and lowers the potential at which it is blocked. With prolonged perfusion, repolarization is blocked at all voltages. When excitability is abolished by perfusion with cobalt, or with a zero calcium solution containing EGTA, no delayed rectification occurs. We suggest that repolarization during the action potential depends on an influx of calcium into the cytoplasm, and that the rate of repolarization depends on the magnitude of the inward calcium current. Increasingly large stimuli reduce the rate of repolarization by reducing the driving force for calcium, and then block repolarization by causing the lumenal membrane potential to exceed ECa. Changes in extracellular calcium affect repolarization in a manner consistent with the resulting change in ECa.     

Central neuronal pathways of the cardioinhibitory reflex in the isopod crustacean Bathynomus doederleini

We have already identified central neurons for cardioinhibition and cardioacceleration in Bathynomus, an isopod crustacean. The 1st thoracic ganglion (TG1) has cardioinhibitory neurons, which we call CIs, while the 2nd and 3rd thoracic ganglia (TG2 and TG3) have cardioacceleratory neurons, which we call CA1s and CA2s. We examined neuronal pathways for cardioinhibitory reflexes in whole animal preparations, using intracellular and extracellular recording methods. Cardiac inhibition in response to a variety of external stimuli was mediated by activation of CIs and inhibition of both CAs. When preparations had the ventral nerve cord intact, CIs were activated by excitatory postsynaptic potentials and CAs were inhibited by inhibitory postsynaptic potentials in response to tactile stimuli applied to sensilla setae on appendages and afferent stimuli applied to ganglionic roots of the thoracic ganglia. However, stimulation of ganglionic nerve roots of TG2 and TG3, or tactile stimulation of the body surface, failed to evoke inhibition of CAs in preparations in which both the cerebral ganglion and TG1 had been excised. These results suggest that TG1 is an indispensable central region for the excitation of CI and for inhibition of CA neurons, induced by tactile stimuli and by stimuli applied to nerve roots of TG2 and TG3.