Excitatory synapses in the glomerular triad of frog olfactory bulb in vitro

Whole-cell patch clamp recording techniques were applied to periglomerular (PG) cells in slices of the frog olfactory bulb (OB) to study the properties of the excitatory synapses in the triad formed by the olfactory nerve (ON) and the dendrites of mitral/tufted (MT) cells and PG cells. The postsynaptic response evoked by ON stimulation was glutamatergic and could be dissected into NMDA and non-NMDA components of equivalent amplitudes. The dendro-dendritic synapse between MT and PG cells could be activated following antidromic stimulation of the lateral and medial olfactory tract (LOT and MOT). In this case the postsynaptic potentials had amplitudes and durations comparable to those obtained by ON stimulation, the neurotransmitter was glutamate, but the synapse was largely dominated by the slow NMDA component.     

Three types of reticular neurons involved in the spino-bulbo-spinal reflex of cats

Reticular neuron activity was recorded in 28 chloralosed cats in order to analyze the reflex arc of the spino-bulbo-spinal (SBS) reflex. Three types of reticular neurons, types I (input), II(output) and III (relay), were identified by unit discharges in response to stimulation of the sural nerve. (1) Type I (input) neurons received spinal ascending volleys monosynaptically and responded to stimulation of the sural nerve with spikes of low amplitude and short latency. Unit spikes, however, were not produced by stimulation of the superficial radial nerve and the sensorimotor cortex. These input neurons were located in the dorsocaudal part of the medial bulbar reticular formation. (2) Type II (output) neurons were part of the reticulospinal tract, which sends axons to the spinal cord, since these neurons exhibited antidromic spikes following stimulation of the ventrolateral funiculus of the spinal cord. Unit spikes were evoked by stimulation either to the sural or superficial radial nerves. These neurons were located in the ventrocaudal part of the medial bulbar reticular formation. (3) Type III neurons included relay neurons. Unit spikes were evoked by stimulation of the sural nerve, superficial radial nerve and sensorimotor cortex. However, unit discharges were not obtained by antidromic stimulation to the reticulospinal tract. These neurons were distributed widely in the brain stem, both in the bulb and pons. (4) Latency difference of unit discharges between input and output neurons was 3.5--5 msec, indicating the presence of interneurons (relays) between input and output neurons. Spikes of output neurons with 3.8--4.2 msec latency were observed following stimulation of the region where input neuron activity was found. We may conclude that three kinds of reticular neurons, input, relay and output, were involved in pathways of the SBS reflex.     

Synaptic organization and neurotransmitters in the rat accessory olfactory bulb

The accessory olfactory bulb (AOB) is the first relay station in the vomeronasal system and may play a critical role in processing pheromone signals. The AOB shows similar but less distinct lamination compared with the main olfactory bulb (MOB). In this study, synaptic organization of the AOB was analyzed in slice preparations from adult rats by using both field potential and patch-clamp recordings. Stimulation of the vomeronasal nerve (VN) evoked field potentials that showed characteristic patterns in different layers of the AOB. Current source density (CSD) analysis of the field potentials revealed spatiotemporally separated loci of inward current (sinks) that represented sequential activation of different neuronal components: VN activity (period I), synaptic excitation of mitral cell apical dendrites (period II), and activation of granule cells by mitral cell basal dendrites (period III). Stimulation of the lateral olfactory tract also evoked field potentials in the AOB, which indicated antidromic activation of the mitral cells (period I and II) followed by activation of granule cells (period III). Whole cell patch recordings from mitral and granule cells of the AOB supported that mitral cells are excited by VN terminals and subsequently activate granule cells through dendrodendritic synapses. Both CSD analysis and patch recordings provided evidence that glutamate is the neurotransmitter at the vomeronasal receptor neuron; mitral cell synapses and both NMDA and non-NMDA receptors are involved. We also demonstrated electrophysiologically that reciprocal interaction between mitral and granule cells in the AOB is through the dendrodendritic reciprocal synapses. The neurotransmitter at the mitral-to-granule synapses is glutamate and at the granule-to-mitral synapse is gamma-aminobutyric acid. The synaptic interactions among receptor cell terminals, mitral cells, and granule cells in the AOB are therefore similar to those in the MOB, suggesting that processing of chemosensory information in the AOB shares similarities with that in the MOB.     

Synaptology of the olfactory bulb of an elasmobranch fish, Sphyrna tiburo

The ultrastructure of the elasmobranch olfactory bulb was examined in order to determine the synaptology of the olfactory circuitry in the bonnethead shark, Sphyrna tiburo. The compartmentalization of the bulb, together with the lack of mitral cell basal dendrites, suggests a different way of performing lateral communication between mitral cells of the olfactory bulb. The results show that granule cells assume an important role by directly interlinking mitral cells. A corollary of this is the segregation of the input onto the mitral cell dendritic arborization: afferent fibers synapse onto the intraglomerular mitral terminals, whereas most local circuit interactions utilize extraglomerular synapses located on the shafts and the somas of the mitral dendrites. Therefore, the elasmobranch synaptic pattern is different from that of higher vertebrates; This might represent the use of a different neural route to achieve the same processing task.     

Metabolic activation of furosemide to a chemically reactive, hepatotoxic metabolite

(1) Intracellular potentials were recorded from pyramidal tract (PT) and non-pyramidal tract (non-PT) cells of the frontal cortex in urethane-anesthetized rabbits and the effects of electrical stimulation of the lateral hypothalamus (LH) were examined on the ipsilateral side. (2) Latencies of antidromic spikes of PT cells evoked by stimulation of the medullary pyramidal tract (PYR) had a unimodal distribution with the mean at 4.0 msec. The mean conduction velocity of the pyramidal tract fibers was 10.5 m/sec. (3) Prolonged IPSPs were produced in PT and non-PT cells by single shock stimulation of LH (LH-IPSP). They were significantly longer lasting than those produced by PYR stimulation (PYR-IPSP). (4) The latencies of LH-IPSPs ranged from 1.7 to 25.0 msec and were divided into two groups. The latencies of PYR-IPSPs had a unimodal distribution ranging from 2.5 to 30 msec. (5) In a few non-PT cells, a sequence of brief depolarization and prolonged hyperpolarization occurred in response to LH stimulation. (6) In some non-PT cells, EPSPs which occasionally resulted in spike discharges were observed after LH stimulation. However, no excitation of PT cells was observed by LH stimulation.     

The organization of piriform cortex and the lateral olfactory tract following the loss of mitral cells in PCD mice

Homozygous Purkinje Cell Degeneration (PCD) mice exhibit a selective loss of olfactory bulb mitral cells (MCs) after 4 months of age. This selective degeneration leaves a subpopulation of denervated granule cells which establish new reciprocal dendro-dendritic synapses with unaffected tufted cells (TCs) (14). This suggests a capacity for plasticity in TCs and raises the question of whether a comparable degree of reorganization occurs in their axonal terminals in piriform cortex (PC) following the loss of MCs. Homozygous (experimental) and heterozygous (control) PCD mice were routinely perfused and processed for electron microscopy. A quantitative electron microscopic analysis was performed on radially oriented micrograph montages spanning from the pia into layer II of PC. After MC loss in the experimental animals there was a decrease in density of larger myelinated axons in the lateral olfactory tract (LOT). Myelinated axons in the LOT had a mean cross-sectional diameter of 1.26 +/- 0.04, and 0.81 +/- 0.025 microm in the control and experimental mice, respectively. In superficial layer I of PC, control mice had presynaptic axonal terminals from mitral and tufted cells with characteristic electron lucent (light) profiles establishing asymmetric synapses with pyramidal cell dendrites. In contrast, the experimental mice showed a decrease in electron lucent terminals and a robust increase in electron dense (dark) presynaptic associational terminals. Although the overall synaptic density did not differ between the control and experimental mice (16.40 +/- 0.94 and 18.10 +/- 0.96 synapses/100 microm2, respectively), an overall decrease in the thickness of Layer 1 suggests that the total number of synapses decreases following MC loss. In addition to the apparent increase of associational terminals, the diameter of terminal enlargements increased as well as the number of multiple synaptic contact per terminals in the experimental animal, suggesting further compensatory mechanisms for the loss of MC presynaptic terminals.     

Beta-galactosidase-labelled relay neurons of homotopic olfactory bulb transplants establish proper afferent and efferent synaptic connections with host neurons

The vertebrate olfactory system has long been an attractive model for studying neuronal regeneration and adaptive plasticity due to the continuous neurogenesis and synaptic remodelling throughout adult life in primary and secondary olfactory centres, its precisely ordered synaptic network and accessibility for manipulation. After homotopic transplantation of fetal olfactory bulbs in bulbectomized neonatal rodents, newly regenerated olfactory neurons form glomeruli within the graft, and the efferent mitral/tufted cells of the transplant innervate the host brain, terminating in higher olfactory centres. However, the synaptic connections of the transplanted relay neurons within the graft and/or host's olfactory centres could not be characterized mainly because of lack of suitable cell-specific markers for these neurons. In this study, we have used olfactory bulbs from transgenic fetuses, in which the majority of the mitral/tufted cells express the bacterial enzyme beta-galactosidase, for homotopic olfactory bulb transplantation following complete unilateral bulbectomy. In the transplants, the cell bodies and terminals of the donor mitral/tufted cells were identified by beta-galactosidase histochemistry and immunocytochemistry at both light and electron microscope levels. We demonstrate that transplanted relay neurons re-establish specific synaptic connections with host neurons of the periphery, source of the primary signal and central nervous system, thereby providing the basis for a functional recovery in the lesioned olfactory system.     

DNA adducts in human placenta as related to air pollution and to GSTM1 genotype

The dendrites of neocortical neurons have been shown to support active action potentials which back-propagate from the soma after an output spike has been initiated. This observation has led to speculation that dendritic action potentials may participate in various forms of synaptic plasticity. The contribution of dendritic spikes to paired-pulse facilitation (PPF), a form of short-term plasticity, was investigated in the dentate gyrus of hippocampal slices. Paired orthodromic stimulation of the perforant path produced an average facilitation of the test population spike (PS) amplitude of 167% (n = 16, conditioning response = 100%). There was also a small but significant increase in slope of the field EPSP (fEPSP) of 108%. To determine whether increased presynaptic drive could account for this facilitation, the relationship between fEPSP slope and spike amplitude (I-O) was determined for a range of stimulus intensities. An increase in fEPSP slope of 171% was associated with an increase in PS amplitude equal to the facilitation produced by paired-pulse stimulation (167%), suggesting a postsynaptic component in PPF. Electric field effects were then used as a tool to alter the excitability of granule cells during the conditioning response without changing synaptic drive. Any change in the test response associated with manipulation of the conditioning population spike amplitude would suggest that dendritic spikes may contribute to the postsynaptic component of PPF. Surprisingly, altering the number of neurons responding to the conditioning stimulus with an action potential had no effect on the test response, suggesting that dendritic action potentials do not participate in this form of short-term synaptic plasticity.     

delta9-tetrahydrocannabinol and the hippocampus: effects on CA1 field potentials in rats

The effects of delta9-tetrahydrocannabinol (THC) on ortho- and antidromically elicited CA1 field potentials were observed in locally anesthetized rats and in anesthetized with urethane. THC augmented amplitudes of population EPSP's as well as orthodromic and antidromic population spikes from pyramidal cells in locally anesthetized animals. Latencies to peak amplitude of these response were increased. Conditioning-test shock experiments revealed taht THC also depressed recurrent inhibition probably mediated by basket cells. In animals under urethane anesthesia THC enhanced test responses, but failed to augment population response to the conditioning stimulus. It was concluded that THC enhanced postsynaptic excitatory processes but attenuated recurrent inhibition. Urethane anesthesia completely blocked the postsynaptic excitatory effect of THC but had little apparent influence on THC's disinhibitory action.     

Conditioning of cortical neurons in cats with antidromic activation as the unconditioned stimulus

Single-cell activity was recorded from the postcruciate cortex of acutely prepared cats during a differential classical conditioning procedure. The conditioned stimuli (CS) were hind paw stimuli, and the unconditioned stimulus (US) was pyramidal tract stimulation that produced an antidromic response in the recorded cortical neuron. A control group was also examined in which the pyramidal stimulus was set below the threshold to produce an antidromic response. Clear differential conditioning was found for the experimental group, with antidromic activation of the neuron as the US. There was no evidence of differential conditioning in the control group without antidromic activation. Any activation of orthodromic pathways should have been the same in the control and experimental groups. The absence of conditioning in the control group demonstrated that orthodromic pathways were not contributing to the differential conditioning observed in the experimental group. This indicates that it was activation of the neuron produced by antidromic firing which was important for conditioning. All the evidence suggests that the site of learning was in the cortex. It is concluded that the the role of the US in conditioning may be simply to activate the neuron at an appropriate interval following the CS.     

Compartmentalization of information processing in an aplysia feeding circuit interneuron through membrane properties and synaptic interactions

We describe a pair of cerebral-to-buccal interneurons, CBI-5/6, which have outputs and inputs in two ganglia. The soma in the cerebral ganglion received synaptic inputs during buccal motor programs (BMPs) and after mechanical stimulation of the lips. During BMPs the soma received antidromic spikes generated in processes in the buccal ganglion. The soma was driven into a plateau potential by each of these inputs, during which it fired orthodromically at 0-5 Hz. The soma had outputs in the cerebral ganglion consisting of electrical coupling to the adjacent CBI-5/6 and to a cerebral-to-pedal neuron (CPN1). The buccal terminals of CBI-5/6 received inputs that generated rhythmic barrages (up to 25 Hz) of antidromic spikes during BMPs. The buccal terminals had chemical and electrical outputs to motor and premotor elements of feeding circuitry. This combination of synaptic interactions and endogenous properties mean that CBI-5/6 can process information in a number of ways. During the barrage of antidromic spikes, high-frequency firing will produce strong inputs to buccal followers and on their arrival at the soma will transfer excitation electrotonically to CPN1. Subthreshold input to the soma will be transferred electrotonically to cerebral followers but will not be relayed to postsynaptic buccal neurons. Plateau potentials after the antidromic spikes or local cerebral inputs will locally excite CPN1 via electrical coupling but will have little influence on buccal events because of the low orthodromic firing rate. Thus, CBI-5/6 may transmit information locally within the cerebral ganglion or more extensively in both buccal and cerebral ganglia simultaneously.     

Mechanisms of excitation and inhibition in the nigrostriatal system

The extracellular responses of neurones in the neostriatum following single pulse stimulation of the substantia nigra were investigated in urethane anaesthetized rats. Low intensity stimulation (less than 10 V) evoked single large amplitude spikes while higher intensities (10-20 V) elicit a high frequency burst of small amplitude spikes or waves. When spontaneous or glutamate-induced large spikes are recorded, nigral stimulation causes their inhibition coincidentally with the development of a burst. If the burst is prevented, the inhibitory response disappears. Both the nigral evoked inhibition and burst response are unaffected by iontophoretically or systemically administered antonists of dopamine or by chemical lesions of the dopamine-containing nigral neurones. The monosynaptic activation of large amplitude striatal neurones, which could also be identified antidromically by stimulation of the globus pallidus, was reversibly blocked by dopamine antagonists. It is concluded (a) that the burst responses are induced through the antidromic excitation of striatonigral axons within the striatum; (b) that the striatal neurones thus activated are inhibitory interneurones and (c) that the dopamine-containing neurones of the nigra make excitatory synaptic contact with a population of striatal output cells, some of which at least project to the globus pallidus.     

The effect of calcium removal on the suppression by adenosine of epileptiform activity in the hippocampus: demonstration of desensitization

1. Previous work has suggested that presynaptic effects of adenosine may be dependent on divalent cations. The present study was undertaken to determine whether a similar requirement existed at postsynaptic sites. 2. Extracellular recordings were made in the CA1 pyramidal cell layer of rat hippocampal slices following orthodromic stimulation of Schaffer collateral fibres in stratum radiatum or antidromic stimulation of the alveus. In antidromic stimulation experiments, CaCl2 was omitted (calcium-free medium) or reduced to 0.24 mM (low calcium medium) and in some experiments MgSO4 was increased to 2 mM. Kynurenic acid at concentrations of 1 and 5 mM in calcium-free medium and 1 mM in low calcium medium had no effect on secondary spike size. 3. Adenosine and baclofen induced a concentration-dependent reduction in the amplitude of orthodromic potentials with maximum effects at 20 and 5 microM respectively. 4. In nominally calcium-free medium, bursts of multiple population spikes were obtained in response to antidromic stimulation. Adenosine had little effect in reducing the secondary spike amplitude. At high concentration (2 mM) an initial depression was seen which declined within 3-5 min. 5. Sensitivity to adenosine was restored in low calcium medium or by raising magnesium. Although raising the divalent cation concentration increased the inhibitory effect of adenosine, desensitization was still seen. 6. 2-Chloroadenosine (100-500 microM) and R-PIA (50 microM), which are not substrates for either the nucleoside transporters or adenosine deaminase, were inactive in the absence of calcium. S-(2-hydroxy-5 nitrobenzyl)-6-thioinosine, an adenosine uptake blocker, at a concentration 100 MicroM had no effect on secondary potential size and did not restore adenosine sensitivity in calcium-free medium.7. Thapsigargin, which discharges intracellular calcium stores, had no significant effect at 1 MicroM on the bursts of action potentials and did not change the effect of 0.5 mM adenosine in calcium-free medium.8. Unlike adenosine, baclofen concentration-dependently reduced the secondary spike size in calcium free medium and no sign of recovery was observed during maintained superfusion for up to 45 min. No cross-desensitization was seen between baclofen and adenosine.9. Applications of adenosine locally by pressure to neuronal somata or dendrites still resulted in desensitized responses in calcium-free medium.10. It is concluded that the postsynaptic sensitivity to adenosine is dependent on the concentration of divalent cations in the extracellular space implying an effect of cations on adenosine receptor activation or transduction processes.     

Estrogen-sensitive neurons with preoptic projection in the lower brain stem of the female rat

Fifty-one neurons in the ventrolateral part of the medulla oblongata were antidromically activated by electrical stimulation of the suprachiasmatic part of the preoptic area in urethane-anestetized, ovariectomized and estrogen-primed female rats. Two types of antidromic responses were distinguished on the basis of their spike configurations and antidromic spike latencies. One type ("fast spikes") was characterized by a fast and smooth rising phase and a shorter duration of the initial positive deflection. The other type ("slow spikes") had a notch in the rising phase and took a longer time to complete the initial deflection. Mean antidromic spike latency for the fast spikes was 9.8 msec while the value for the slow spikes was 30.2 msec. Ionophoretic injection of estradiol was accomplished on 37 of the 51 antidromically identified cells, of which 21 showed slow responses and 16 responded with fast spikes. In cells with slow spikes, estradiol facilitated (n = 9) or suppressed (n = 3) their generation of action potentials. None of cells with fast responses changed their activity in response to estradiol. It is evident from the present experiment that neurons in the ventrolateral part of the medulla oblongata send their axons directly to the suprachiasmatic part of the preoptic area which plays an important role in the control of the ovulatory surge of LH and that some of these neurons themselves are the sensitive sites of estradiol.     

Response plasticity in hamster olfactory bulb: peripheral and central processes

It is a well-established fact that prolonged odor stimulation leads to marked sensory adaptation. This study demonstrates comparable electrophysiological phenomena occurring at the level of the olfactory receptor and at more central olfactory structures. Recordings of overall receptor response and of olfactory bulb unit responses were made during repeated odor stimulation. During the course of a single, continuous odor presentation response decrements were seen in the EOG (at the olfactory receptors) and were mirrored at the mitral cell layer of the bulb. When brief periods without stimulation were introduced between such odor presentations, receptor responsiveness rebounded to its original level, but mitral cell responses did not. On the basis of this dissociation it is suggested that the pattern of response decrement within the bulb represents a case of stimulus-specific habituation in a simple cortical subsystem and is well worth future investigation as a model of neural plasticity. Surgical disconnection of the olfactory bulb from one or more of its centrifugal inputs results in hyperactive, hyperresponsive mitral cells, which habituate more rapidly and show longer recovery times than do those in the intact bulb. In addition, the synchronization of such units to the inhalation cycle is markedly reduced as compared with normal preparations. These facts together suggest that the habituation of mitral cell activity does not depend on centrifugal inputs, although one or more of such inputs act indirectly in an inhibitory fashion to modulate and tune mitral cell response characteristics.     

Inhibitory control of somatodendritic interactions underlying action potentials in neocortical pyramidal neurons in vivo: an intracellular and computational study

The effect of synaptic inputs on somatodendritic interactions during action potentials was investigated, in the cat, using in vivo intracellular recording and computational models of neocortical pyramidal cells. An array of 10 microelectrodes, each ending at a different cortical depth, was used to preferentially evoke synaptic inputs to different somatodendritic regions. Relative to action potentials evoked by current injection, spikes elicited by cortical microstimuli were reduced in amplitude and duration, with stimuli delivered at proximal (somatic) and distal (dendritic) levels evoking the largest and smallest decrements, respectively. When the inhibitory postsynaptic potential reversal was shifted to around -50 mV by recording with KCl pipettes, synaptically-evoked spikes were significantly less reduced than with potassium acetate or cesium acetate pipettes, suggesting that spike decrements are not only due to a shunt, but also to voltage-dependent effects. Computational models of neocortical pyramidal cells were built based on available data on the distribution of active currents and synaptic inputs in the soma and dendrites. The distribution of synapses activated by extracellular stimulation was estimated by matching the model to experimental recordings of postsynaptic potentials evoked at different depths. The model successfully reproduced the progressive spike amplitude reduction as a function of stimulation depth, as well as the effects of chloride and cesium. The model revealed that somatic spikes contain an important contribution from proximal dendritic sodium currents up to approximately 100 microm and approximately 300 microm from the soma under control and cesium conditions, respectively. Proximal inhibitory postsynaptic potentials can present this dendritic participation thus reducing the spike amplitude at the soma. The model suggests that the somatic spike amplitude and shape can be used as a "window" to infer the electrical participation of proximal dendrites. Thus, our results suggest that inhibitory postsynaptic potentials can control the participation of proximal dendrites in somatic sodium spikes.     

Production of interferon-gamma in families with multiple occurrence of schizophrenia

The effects of the two structurally related Aconitum alkaloids, 1-benzoylnapelline and napelline, were investigated by extracellular recording of the stimulus-evoked population spike in the CA1 region of rat hippocampal slices in vitro. 1-Benzoylnapelline (1-100 microM) exerted a depressant action on the orthodromic as well as on the antidromic population spike. Napelline failed to evoke a significant effect at concentrations up to 100 microM. The inhibitory action induced by 1-benzoylnapelline was enhanced when the frequency of electrical stimulation was increased. In contrast, reversal of the inhibitory effect was accelerated when stimulation frequency was decreased. The activity-dependent mode of action of 1-benzoylnapelline raised the question of whether the drug is effective to suppress epileptiform activity. The results obtained from experiments on epileptiform hippocampal slices revealed a reduction of the burst duration and of the number of spikes in the burst as well as attenuation of the amplitude of the population spikes. These data support the conclusion that 1-benzoylnapelline, in contrast to the structurally related compound, napelline, has an activity-dependent inhibitory action on central neurons.     

Responses of cells of posterior lateral line lobe to activation of electroreceptors in a mormyrid fish

Activity of neurons in the lateral line lobe was studied by intracellular recording of responses to stimulation of the lateral line nerves and of electroreceptors on the skin surface. Two modes of activation occur for cells responding to inputs from medium receptors. There is a direct monosynaptic input mediated by a single fiber. Short latency of response and antidromic spread from cell to afferent fiber indicate that the mediating synapse is electrotonic. The second input is from a number of additional fibers and is relayed, presumably by the granule cells. At shortest latency this input is disynaptic, probably involving at least one electrotonic synapse. A relay is indicated by heterosynaptic facilitation of the PSP and by pronounced depression with repetitive stimulation. The monosynaptic input may be on the axon. Disynaptic inputs are distributed over the dendrites, and impulses can arise in the dendrites. What appear to be spikes restricted to dendritic regions are often recorded as small brief potentials in the cell body. There is a somatotopic projection of the electroreceptors to the lateral line lobe. The monosynaptic input comes from a specific receptor in the periphery. Strong disynaptic inputs come from a group of receptors generally found anterior, but less commonly posterior or lateral, to the receptor giving rise to the monosynaptic input. Additional inputs that are inhibitory come from surrounding receptors. The inhibition only affects responses to the disynaptic input. The different inputs and multiple sites of impulse initiation must modify the cell's response as compared with the input-output relations that would be obtained with inputs acting on a single summation point. Cells responding to activation of large receptors are infrequent. They are characterized by low threshold, little latency change near threshold, and ability to follow high frequencies of stimulation.