Sites Of Synaptic Input Research Articles

Batteries and the brain

As a result of the work of Galvani, Volta, Purkyne, Hitzig and others over 150 years ago, we know that weak electrical currents can exert potent effects on neural tissue in animals and humans. They can polarise neuronal membranes and modulate the firing of neurones as in Galvanic stimulation of the vestibular system (Goldberg et al. 1984). Development of transcranial direct current stimulation (tDCS) as a research tool and as a potential therapy in neurology that is both simple and safe has focused attention on evidence of how it modulates brain activity. Delivery of direct current (DC) between two large electrodes placed on the head produces long-lasting changes in the CNS that greatly outlast the stimulus. These have variably been shown to alter motor functions (e.g. skill learning, recovery of function following stroke) and also sensory functions. Changes in cognitive function and mood have also attracted attention. Critical issues are what mechanisms underlie these changes and how might they be harnessed reliably. It is usually assumed that tDCS acts on the cerebral cortex, and plastic changes in cortical networks are frequently invoked to explain its long-lasting after-effects. In this issue, Bolzoni et al. (2013) tackle this assumption for motor function and, by extrapolation, for many other functions. In studies in anaesthetised cats, they found that tDCS also produces long-lasting changes in the ‘excitability’ of descending motor pathways that originate in the brainstem brought about by tDCS. This was established using well-understood methods to evoke responses in descending motor pathways which were recorded as compound action potentials from the spinal cord en route to spinal motor nuclei. An important feature is that electrical pulses delivered via in-dwelling extracellular electrodes can be adjusted to activate descending motor pathways trans-synaptically (via activation of terminals presynaptic to the cells with descending axons) and/or directly at the soma/initial segment or descending axon (which usually requires higher stimulus intensities). In each case, the trans-synaptic and directly activated responses can be unambiguously differentiated. All descending paths which were tested (including rubrospinal, reticulospinal and vestibulospinal paths) showed increased trans-synaptic activation following tDCS. The increased responsiveness developed gradually during application of the DC stimulus and greatly outlasted it, a time course observed in human studies. Thus, it is possible that some of the motor effects described in humans include effects mediated ‘remotely’ by brainstem descending motor pathways and that the effects are not exclusively mediated via corticospinal or corticofugal pathways. Changes ascribed to tDCS in other systems may therefore also be mediated via non-cortical paths. Regarding the mechanisms, these robust increases in indirect activation of rubrospinal and reticulospinal neurons following tDCS (rubrospinal probably via cerebellar efferents and reticulospinal from intrinsic reticular or corticoreticular terminals) were also from pathways that are not commonly associated with plasticity, and occurred during anaesthesia. In addition, tDCS sometimes increased output from directly activated axons, including reticulospinal and corticospinal tracts, axons of which were activated in the medulla some distance from sites of synaptic input. Thus, some effects of tDCS may be exerted on axons and therefore be unrelated to synaptic mechanisms, and therefore unrelated to long-term plastic changes. There is ample evidence from animal studies that the precise orientation of the dendritic fields to the current path will influence the mode and magnitude of neuronal excitation (e.g. Hern et al. 1962; Kabakov et al. 2012) and that tDCS can also alter presynaptic function (e.g. Marquez-Ruiz et al. 2012). An issue that needs to be resolved is to what extent the changes described in this study might also operate in humans. Observations in such different species must involve some differences: for example the smaller skull of the cat precludes the use of similarly sized electrodes to those used in the human studies and the different proportional brain anatomy must influence the current paths. The current densities in the study in the cat were probably at least twice those used in human studies. Not surprisingly there were also differences in the time course of effects. A remarkable feature of the work is that depressions in excitability were not reported, whereas the polarity and cranial localization of electrodes used to deliver the tDCS appear critical to its effectiveness in humans. To what extent there are common cellular features underlying the effects of DC stimulation at cortical, subcortical and spinal sites remains to be resolved.

Electrical Advantages of Dendritic Spines

Many neurons receive excitatory glutamatergic input almost exclusively onto dendritic spines. In the absence of spines, the amplitudes and kinetics of excitatory postsynaptic potentials (EPSPs) at the site of synaptic input are highly variable and depend on dendritic location. We hypothesized that dendritic spines standardize the local geometry at the site of synaptic input, thereby reducing location-dependent variability of local EPSP properties. We tested this hypothesis using computational models of simplified and morphologically realistic spiny neurons that allow direct comparison of EPSPs generated on spine heads with EPSPs generated on dendritic shafts at the same dendritic locations. In all morphologies tested, spines greatly reduced location-dependent variability of local EPSP amplitude and kinetics, while having minimal impact on EPSPs measured at the soma. Spine-dependent standardization of local EPSP properties persisted across a range of physiologically relevant spine neck resistances, and in models with variable neck resistances. By reducing the variability of local EPSPs, spines standardized synaptic activation of NMDA receptors and voltage-gated calcium channels. Furthermore, spines enhanced activation of NMDA receptors and facilitated the generation of NMDA spikes and axonal action potentials in response to synaptic input. Finally, we show that dynamic regulation of spine neck geometry can preserve local EPSP properties following plasticity-driven changes in synaptic strength, but is inefficient in modifying the amplitude of EPSPs in other cellular compartments. These observations suggest that one function of dendritic spines is to standardize local EPSP properties throughout the dendritic tree, thereby allowing neurons to use similar voltage-sensitive postsynaptic mechanisms at all dendritic locations.

B56β, a regulatory subunit of protein phosphatase 2A, interacts with CALEB/NGC and inhibits CALEB/NGC‐mediated dendritic branching

The development of dendritic arbors is critical in neuronal circuit formation, as dendrites are the primary sites of synaptic input. Morphologically specialized dendritic protrusions called spines represent the main postsynaptic compartment for excitatory neurotransmission. Recently, we demonstrated that chicken acidic leucine-rich epidermal growth factor (EGF) -like domain-containing brain protein/neuroglycan C (CALEB/NGC), a neural member of the EGF family, mediates dendritic tree and spine complexity but that the signaling pathways in the respective processes differ. For a more detailed characterization of these signal transduction pathways, we performed a yeast two-hybrid screen to identify proteins that interact with CALEB/NGC. Our results show that B56beta, a regulatory subunit of protein phosphatase 2A, interacts with CALEB/NGC and inhibits CALEB/NGC-mediated dendritic branching but not spine formation. Binding of B56beta to CALEB/NGC was confirmed by several biochemical and immunocytochemical assays. Using affinity chromatography and mass spectrometry, we demonstrate that the whole protein phosphatase 2A trimer, including structural and catalytic subunits, binds to CALEB/NGC via B56beta. We show that CALEB/NGC induces the phosphorylation of Akt in dendrites. Previously described to interfere with Akt signaling, B56beta inhibits Akt phosphorylation and Akt-dependent dendritic branching but not Akt-independent spine formation induced by CALEB/NGC. Our results contribute to a better understanding of signaling specificity leading to neuronal process differentiation in sequential developmental events.

The neural EGF family member CALEB/NGC mediates dendritic tree and spine complexity

The development of dendritic arborizations and spines is essential for neuronal information processing, and abnormal dendritic structures and/or alterations in spine morphology are consistent features of neurons in patients with mental retardation. We identify the neural EGF family member CALEB/NGC as a critical mediator of dendritic tree complexity and spine formation. Overexpression of CALEB/NGC enhances dendritic branching and increases the complexity of dendritic spines and filopodia. Genetic and functional inactivation of CALEB/NGC impairs dendritic arborization and spine formation. Genetic manipulations of individual neurons in an otherwise unaffected microenvironment in the intact mouse cortex by in utero electroporation confirm these results. The EGF-like domain of CALEB/NGC drives both dendritic branching and spine morphogenesis. The phosphatidylinositide 3-kinase (PI3K)-Akt-mammalian target of rapamycin (mTOR) signaling pathway and protein kinase C (PKC) are important for CALEB/NGC-induced stimulation of dendritic branching. In contrast, CALEB/NGC-induced spine morphogenesis is independent of PI3K but depends on PKC. Thus, our findings reveal a novel switch of specificity in signaling leading to neuronal process differentiation in consecutive developmental events.

Learning Rules for Spike Timing-Dependent Plasticity Depend on Dendritic Synapse Location

Previous studies focusing on the temporal rules governing changes in synaptic strength during spike timing-dependent synaptic plasticity (STDP) have paid little attention to the fact that synaptic inputs are distributed across complex dendritic trees. During STDP, propagation of action potentials (APs) back to the site of synaptic input is thought to trigger plasticity. However, in pyramidal neurons, backpropagation of single APs is decremental, whereas high-frequency bursts lead to generation of distal dendritic calcium spikes. This raises the question whether STDP learning rules depend on synapse location and firing mode. Here, we investigate this issue at synapses between layer 2/3 and layer 5 pyramidal neurons in somatosensory cortex. We find that low-frequency pairing of single APs at positive times leads to a distance-dependent shift to long-term depression (LTD) at distal inputs. At proximal sites, this LTD could be converted to long-term potentiation (LTP) by dendritic depolarizations suprathreshold for BAC-firing or by high-frequency AP bursts. During AP bursts, we observed a progressive, distance-dependent shift in the timing requirements for induction of LTP and LTD, such that distal synapses display novel timing rules: they potentiate when inputs are activated after burst onset (negative timing) but depress when activated before burst onset (positive timing). These findings could be explained by distance-dependent differences in the underlying dendritic voltage waveforms driving NMDA receptor activation during STDP induction. Our results suggest that synapse location within the dendritic tree is a crucial determinant of STDP, and that synapses undergo plasticity according to local rather than global learning rules.

Boosting of action potential backpropagation by neocortical network activity in vivo.

Action potentials backpropagate into the dendritic trees of pyramidal neurons, reporting output activity to the sites of synaptic input and provoking long-lasting changes in synaptic strength. It is unclear how this retrograde signal is modified by neural network activity. Using whole-cell recordings from somata, apical trunks, and dendritic tuft branches of layer 2/3 pyramidal neurons in vivo, we show that network-driven subthreshold membrane depolarizations ("up states") occur simultaneously throughout the apical dendritic tree. This spontaneous synaptic activity enhances action potential-evoked calcium influx into the distal apical dendrite by promoting action potential backpropagation. Hence, somatic feedback to the dendrites becomes stronger with increasing network activity.

NMDA Receptor-Mediated Na+Signals in Spines and Dendrites

Spines and dendrites of central neurons represent an important site of synaptic signaling and integration. Here we identify a new, synaptically mediated spine signal with unique properties. Using two-photon Na(+) imaging, we show that suprathreshold synaptic stimulation leads to transient increases in Na(+) concentration in postsynaptic spines and their adjacent dendrites. This local signal is restricted to a dendritic domain near the site of synaptic input. In presumed active spines within this domain, the Na(+) level increases by 30-40 mm even during short bursts of synaptic stimulation. During a long-term potentiation induction protocol (100 Hz, 1 sec), the Na(+) level in the active spines reaches peak amplitudes of approximately 100 mm. We find that the Na(+) transients are mainly mediated by Na(+) entry through NMDA receptor channels and are detected during the coincident occurrence of synaptic potentials and backpropagating action potentials. The large amplitudes of the Na(+) transients and their location on dendritic spines suggest that this signal is an important determinant of electrical and biochemical spine characteristics.

Passive electrotonic properties of rat hippocampal CA3 interneurones.

1. The linear membrane responses of CA3 interneurones were determined with the use of whole-cell patch recording methods. The mean input resistance (RN) for all cells in this study was 526 +/- 16 MOmega and the slowest membrane time constant (tau0) was 73 +/- 3 ms. 2. The three-dimensional morphology of 63 biocytin-labelled neurones was used to construct compartmental models. Specific membrane resistivity (Rm) and specific membrane capacitance (Cm) were estimated by fitting the linear membrane response. Acceptable fits were obtained for 24 CA3 interneurones. The mean Rm was 61.9 +/- 34.2 Omega cm2 and the mean Cm was 0.9 +/- 0.3 microF cm-2. Intracellular resistance (Ri) could not be resolved in this study. 3. Examination of voltage attenuation revealed a significantly low synaptic efficiency from most dendritic synaptic input locations to the soma. 4. Simulations of excitatory postsynaptic potentials (EPSPs) were analysed at both the site of synaptic input and at the soma. There was little variability in the depolarization at the soma from synaptic inputs placed at different locations along the dendritic tree. The EPSP amplitude at the site of synaptic input was progressively larger with distance from the soma, consistent with a progressive increase in input impedance. 5. The 'iso-efficiency' of spatially different synaptic inputs arose from two opposing factors: an increase in EPSP amplitude at the synapse with distance from the soma was opposed by a nearly equivalent increase in voltage attenuation. These simulations suggest that, in these particular neurones, the amplitude of EPSPs measured at the soma will not be significantly affected by the location of synaptic inputs.

A synaptically controlled, associative signal for Hebbian plasticity in hippocampal neurons.

The role of back-propagating dendritic action potentials in the induction of long-term potentiation (LTP) was investigated in CA1 neurons by means of dendritic patch recordings and simultaneous calcium imaging. Pairing of subthreshold excitatory postsynaptic potentials (EPSPs) with back-propagating action potentials resulted in an amplification of dendritic action potentials and evoked calcium influx near the site of synaptic input. This pairing also induced a robust LTP, which was reduced when EPSPs were paired with non-back-propagating action potentials or when stimuli were unpaired. Action potentials thus provide a synaptically controlled, associative signal to the dendrites for Hebbian modifications of synaptic strength.

Ultrastructural Immunocytochemical Localization of μ-Opioid Receptors in Rat Nucleus Accumbens: Extrasynaptic Plasmalemmal Distribution and Association with Leu5-Enkephalin

mu-Opioid receptors and their endogenous ligands, including Leu5-enkephalin (LE), are distributed abundantly in the nucleus accumbens (NAC), a region implicated in mechanisms of opiate reinforcement. We used immunoperoxidase and/or immunogold-silver methods to define ultrastructural sites for functions ascribed to mu-opioid receptors and potential sites for activation by LE in the NAC. An antipeptide antibody raised against an 18 amino acid sequence of the cloned mu-opioid receptor (MOR) C terminus showed that MOR-like immunoreactivity (MOR-LI) was localized predominantly to extrasynaptic sites along neuronal plasma membranes. The majority of neuronal profiles containing MOR-LI were dendrites and dendritic spines. The dendritic plasma membranes immunolabeled for MOR were near sites of synaptic input from LE-labeled terminals and other unlabeled terminals forming either inhibitory or excitatory type synapses. Unmyelinated axons and axon terminals were also intensely but less frequently immunoreactive for MOR. Observed sites for potential axonal associations with LE included coexistence of MOR and LE within the same terminal, as well as close appositions between differentially labeled axons. Astrocytic processes rarely contained detectable MOR-LI, but also were sometimes observed in apposition to LE-labeled terminals. We conclude that in the rat NAC, MOR is localized prominently to extrasynaptic neuronal and more rarely to glial plasma membranes that are readily accessible to released LE and possibly other opioid peptides and opiate drugs. The close affiliation of MOR with spines receiving excitatory synapses and dendrites receiving inhibitory synapses provides the first direct morphological evidence that MOR selectively modulates postsynaptic responses to cortical and other afferents.

Action in the Dendrites: A Revisitation of Dendritic Action Potentials

Dendntes are the primary site of synaptic input to neurons in the CNS. They are imbued with specialized structures, such as dendritic spines, which receive much of the synaptic inputs from other neurons. They have long been considered electrically passive in that they receive nonregenerative synaptic inputs, both excitatory and inhibitory, and, through summation of passive electrotonic spread of current, they convey information in the form of membrane potential changes to the cell body and the initial axon segment, which are specialized to initiate action potentials. More than 40 years ago, this view was challenged by electrophysiological studies suggesting that dendrites can also generate action po tentials. It was proposed that these regenerative dendritic action potentials can serve as "booster" sites to assure the fidelity of synaptic signaling to the distant initial axon seg ment where nerve impulses are initiated for propagation to their terminals for signal trans mission to other neurons. Two recent articles in Science reporting the use of simultaneous patch clamp recordings from a dendrite and the cell body of rat hippocampal neurons have rekindled interest in dendrites as active generators of action potentials. In this Up date, these articles are reviewed and discussed in the context of the functional implica tions of active dendritic electrogenesis. The Neuroscientist 1:312-316, 1995

Subthreshold synaptic activation of voltage-gated Ca2+ channels mediates a localized Ca2+ influx into the dendrites of hippocampal pyramidal neurons.

1. Whole cell recordings and high-speed fluorescence imaging were used to investigate the spatial and temporal characteristics of Ca2+ influx during synaptic activity in hippocampal CA1 pyramidal neurons. Brief, subthreshold trains of synaptic potentials elicited by Schaffer collateral stimulation produced transient increases in [Ca2+]i in the apical dendrites near the site of synaptic input. The rises in [Ca2+]i were not due to Ca2+ entry through N-methyl-D-aspartate (NMDA)-activated or non-NMDA-activated glutamate channels, but were reduced by low concentrations of Ni2+. Hyperpolarizing prepulses caused an increase in the synaptically evoked Ca2+ transients, whereas strong hyperpolarization during the train prevented the rise in [Ca2+]i. The data suggest that subthreshold synaptic activity can open low-voltage-activated (T-type) Ca2+ channels and produce a local increase in intradendritic [Ca2+]. Such local increases in [Ca2+]i may be important for modulating the strength of synaptic connections.

A model for dendritic Ca2+ accumulation in hippocampal pyramidal neurons based on fluorescence imaging measurements.

1. High-speed fluorescence imaging was used to measure intracellular Ca2+ concentration ([Ca2+]i) changes in hippocampal neurons injected with the Ca(2+)-sensitive indicator fura-2 during intrasomatic and synaptic stimulation. The results of these experiments were used to construct a biophysical model of [Ca2+]i dynamics in hippocampal neurons. 2. A compartmental model of a pyramidal neuron was constructed incorporating published passive membrane properties of these cells, three types of voltage-gated Ca2+ channels characterized from adult hippocampal neurons, voltage-gated Na+ and K+ currents, and mechanisms for Ca2+ buffering and extrusion. 3. In hippocampal pyramidal neurons imaging of Na+ entry during electrical activity suggests that Na+ channels, at least in sufficient density to sustain action potentials, are localized in the soma and the proximal part of the apical dendritic tree. The model, which incorporates this distribution, demonstrates that action potentials attenuate steeply in passive distal dendritic compartments or distal dendritic compartments containing Ca2+ and K+ channels. This attenuation was affected by intracellular resistivity but not membrane resistivity. 4. Consistent with fluorescence imaging experiments, a non-uniform distribution of Ca2+ accumulation was generated by Ca2+ entry through voltage-gated Ca2+ channels opened by decrementally propagating Na+ action potentials. Consequently, the largest increases in [C2+]i were produced in the proximal dendrites. Distal voltage-gated Ca2+ currents were activated by broad, almost isopotential action potentials produced by reducing the overall density of K+ channels. 5. Simulations of subthreshold synaptic stimulation produced dendritic Ca2+ entry by the activation of voltage-gated Ca2+ channels. In the model these Ca2+ signals were localized near the site of synaptic input because of the attenuation of synaptic potentials with distance from the site of origin and the steep voltage-dependence of Ca2+ channel activation. 6. These simulations support the hypotheses generated from experimental evidence regarding the differential distribution of voltage-gated Ca2+ and Na+ channels in hippocampal neurons and the resulting voltage-gated Ca2+ accumulation from action and synaptic potentials.

Characterization of Ca current underlying burst formation in lobster cardiac ganglion motorneurons

1. The anterior motorneurons of the cardiac ganglion of Homarus americanus were ligated less than 300 microns from the soma. This removes impulse-generating membrane and sites of synaptic input while preserving the ability of the soma to generate the burst-forming potentials termed "driver potentials" regenerative, slow (250-ms duration) depolarizations (to -20 mV) in response to brief, depolarizing stimuli. At stimulus intervals corresponding to rates of bursting observed in spontaneously active, intact ganglia (0.3-1.2/s), driver potential amplitude increases with increasing stimulus interval. 2. A two-electrode voltage clamp was used to characterize inward current observable from the ligated neurons in tetrodotoxin (TTX)-tetraethylammonium (TEA)-containing salines. The amplitude of inward current shows a hyperbolic relation to [Ca]o that is well fitted by a form of the Michaelis-Menten equation. Inward current is maintained but not augmented when Ca2+ is replaced by Ba2+ or Sr2+. It is concluded that the inward current, to be referred to as ICa, is mediated by voltage-dependent Ca channels. 3. Contamination of ICa by early outward current (IA) was evaluated by addition of 4-aminopyridine (4-AP, 4 mM). In the presence of 4-AP, the net inward current is increased and the potential at which maximum ICa occurs is shifted 10 mV more positive. 4. Subtraction of outward currents recorded in Mn2(+)-containing saline from overall currents in the absence of Mn2+ provided another means to separate inward from outward current. I-V curves from such "Mn-subtracted" records show ICa approaches a saturating value for steps to -5 mV and more depolarized. The time to peak ICa is voltage dependent. The largest inward currents (up to 240 nA) and minimal time to peak (4 ms) are observed for steps from holding potentials of -50 to -60 mV. 5. Decline of ICa during depolarized steps observed in Mn-subtracted records represents inactivation rather than development of competing outward current. Inactivation is slow and incomplete; the rate and fractional amount of inactivation are not directly voltage dependent. Nonsubtracted responses to 500-ms depolarizations to potentials evoking little outward current show that an initial rapid decline of ICa (tau approximately 40 ms) is followed at approximately 80 ms by a slower phase of decline (tau approximately 180 ms). With repetitive clamps, the early phase proved labile.(ABSTRACT TRUNCATED AT 400 WORDS)

Sodium channels in dendrites of rat cortical pyramidal neurons.

The voltage-dependent properties that have been directly demonstrated in Purkinje cell and hippocampal pyramidal cell dendrites play an important role in the integrative capacities of these neurons. By contrast, the properties of neocortical pyramidal cell dendritic membranes have been more difficult to assess. Active dendritic conductances near sites of synaptic input would have an important effect on the input-output characteristics of these neurons. In the experiments reported here, we obtained direct evidence for the existence of voltage-dependent Na+ channels on the dendrites of neocortical neurons by using cell-attached patch and whole cell recordings from acutely isolated rat neocortical pyramidal cells. The qualitative and quantitative properties of dendritic and somatic currents were indistinguishable. Insofar as Na+ currents are concerned, the soma and primary apical dendrite can be considered as one relatively uniform compartment. Similar dendritic Na+ currents on dendrites in mature neurons would play an important role in determining the integrative properties of these cortical units.

Real-time imaging of evoked activity in local circuits of the salamander olfactory bulb.

The encoding of olfactory information in the central nervous system (CNS) depends on spatially distributed patterns of activity generated simultaneously in many neuronal circuits. Optical neurophysiological recording permits analysis of neural activity non-invasively and with high spatial and temporal resolution. Here, a video method for imaging voltage-sensitive dye fluorescence in vivo is used to map neuronal activity in local circuits of the salamander olfactory bulb. The method permits the imaging of simultaneous ensemble transmembrane activity in real time. After electrical stimulation of the olfactory nerve, activity spreads centripetally from the sites of synaptic input to generate nonhomogeneous response patterns that are presumably mediated by local circuits within the bulbar layers. The results also show the overlapping temporal sequences of activation of cell groups in each layer. The method thus provides high resolution, sequential video images of the spatial and temporal progression of transmembrane events in neuronal circuits after afferent stimulation and offers the opportunity for studying ensemble events in other brain regions.

Morphology, ultrastructure and synapse distribution of putative serotonergic salivary neurons in the locust

Neurons innervating the locust salivary glands have been investigated using the electron microscope. We have examined the projections in the suboesophageal ganglion of two identified salivary neurons by differential double labelling with cobalt ions, followed by silver intensification. Numerous synaptic inputs occur on the arborizations of the salivary neurons, particularly on fine branches and on small spines arising from larger branches. Although a few instances occur of common input to both salivary neurons from a single presynaptic element, many of the appositions between branches seen in the light microscope do not represent functional connections. A few structures resembling presynaptic dense bodies have been observed in salivary neuron profiles, but with few synaptic vesicles. Large dense granules are present in some labelled profiles, but not in the vicinity of synapse-like membrane specializations. We have also examined unidentified neuron profiles within the acini of the salivary glands, which contain large dense granules and small vesicles. There is good evidence that these unidentified terminals correspond to the suboesophageal salivary neurons. The central arborizations of the salivary neurons appear to serve largely as sites of synaptic input, whereas the peripheral terminals are likely sites of transmitter release. The results are considered in the context of the known immunoreactivity of the salivary neurons with antibodies to 5-hydroxytryptamine, and compared with analagous systems in other insect groups.

Synaptic integration in excitatory and inhibitory crayfish motoneurons.

The passive integrative properties of two crayfish abdominal motoneurons, the fast flexor inhibitor (FI) and a posterior, ipsilateral fast flexor excitor (FE), were studied electrophysiologically and through simulations with multicompartment models of their electrotonic structures. Responses of the models to simulated giant neuron input were quite similar to the motoneurons' responses to giant neuron stimulation, which suggests that differences in the electrotonic structures and the sites of synaptic input to the two cells can account in large part for differences in their responses to a common input. A full action potential created in the initial axon compartment of the FI model produced attenuated potentials in the adjacent integrating segment compartment and contralateral soma compartment. These potentials are similar in amplitude and time course to attenuated antidromic action potentials recorded in the corresponding regions of the FI neuron. A location of the spike initiation zone of the FI at the initial axon segment is consistent with this result. The responses of FI to ipsi- and contralateral inputs are different. Shock of a single abdominal second root produced a larger, faster rising excitatory postsynaptic potential in the ipsilateral FI soma than in the contralateral soma. Second root shock also caused the contralateral FI to produce an action potential either alone or before the ipsilateral FI neuron. Responses of the FI model to ipsilateral and contralateral inputs differ in the same way as the cell's responses. Inputs to the FI model that are ipsilateral to the soma compartment produce larger responses there than do contralateral inputs. Conversely, those contralateral inputs produce larger responses in the initial axon compartment than do ipsilateral inputs. This difference results from the long integrating segment that connects the soma compartment to the initial axon compartment. These results can account for the FI responses to lateralized inputs. Unlike the responses of FIs, the soma responses of contralaterally homologous FEs to ipsilateral and contralateral second root shocks were similar in waveform and amplitude, with the ipsilateral root producing the larger response. This result is consistent with theoretical results from the FE model simulations. We conclude that a smaller size, larger input resistance and shorter membrane time constant allow the FE to respond to giant neuron input before the FI, and so help to achieve the proper timing of flexor contraction and relaxation during a tailflip.(ABSTRACT TRUNCATED AT 400 WORDS)

Calcium and protein kinase C inhibit biosynthesis of Aplysia egg-laying hormone

Previous studies on the biosynthesis of the peptide egg-laying hormone (ELH) of Aplysia have suggested that the increase in cAMP levels associated with the initiation of a bag cell discharge stimulates ELH synthesis, whereas the calcium influx associated with the discharge inhibits it. This report provides additional documentation of the inhibitory role of calcium. Inhibition by the calcium ionophore A23187 was shown to be dependent on the presence of extracellular calcium. A23187 inhibited ELH synthesis and exposure to 0 Ca 2+/2 mM EGTA medium stimulated it in bag cell somata surgically deprived of their sites of synaptic input. The tumor-promoting phorbol ester, TPA, inhibited ELH biosynthesis in a calcium-dependent fashion, whereas the non-tumor-promoting 4α-phorbol did not. These results are consistent with the hypothesis that calcium entry during bag cell discharge may inhibit ELH synthesis via activation of protein kinase C, thus counteracting the stimulation by cAMP early in the discharge. Such a mechanism could precisely regulate the production of ELH molecules to replace those lost by secretion.

Morphology and physiology of auditory interneurons in the metathoracic ganglion of the locust

1. Auditory interneurons in the metathoracic ganglion of the locust were characterized by their intracellularly recorded responses to sound stimuli, and by their morphology as revealed in sectioned preparations of single neurons injected with Lucifer Yellow. Accordingly, nine interneurons were identified as either local, bisegmental, ascending or T-neurons (Figs. 2–6). 2. All cells possess arborizations within one or both sides of a prominent area of fine neuropil (frontal auditory neuropil), but the degree of overlap with the endings of tympanic afferents is different. For 2 ascending units (AN2, AN3) there is no overlap at all, indicating that they are interneurons of higher order. 3. Two neurons, SN1 and TN1, were found with only excitatory input to low frequency sound, and their tuning reflected that one of low frequency receptor fibres. In contrast, the response of all other neurons to low frequency stimuli was more complex, consisting of excitatory and inhibitory synaptic potentials. The neuronal circuitry which might underlie such physiological behaviour is discussed. 4. Recordings made from different parts of individual neurons enabled the sites of synaptic input and output to be localized. The postsynaptic dendrites of the interneurons appear to correspond to smooth endings in contrast to the beaded appearance of the presumptive presynaptic terminals (Fig. 8). 5. The results include descriptions of auditory ‘sister-neurons’ (Fig. 6) which have a similar morphology and of ‘twin-neurons’ (Fig. 7), which, using our present physiological and morphological criteria, cannot reliably be distinguished one from another. The problem that arises from such doubling of cells for the concept of identified units is discussed.