Synaptic activity modulates presynaptic excitability

Synaptic activity modulates synaptic efficacy and is important in learning and development. Here we show that development of excitability in presynaptic motor neurons required synaptic activation of postsynaptic muscle cells. Synaptic blockade broadened action potentials and decreased repetitive firing of presynaptic neurons. Consistent with these findings, synaptic blockade also decreased potassium-current density in the presynaptic cell. Application of neurotrophin-3, but not related neurotrophins, prevented these changes. Recordings from patches of somatic membrane indicated that modifications of presynaptic potassium and sodium currents occurred in a remote, nonsynaptic compartment. Thus, activity-dependent postsynaptic signals modulated presynaptic excitability, potentially regulating transmission at all synapses of the presynaptic cell.     

Synaptic enhancement induced by NMDA and Qp receptors and presynaptic activity.

Activity-dependent changes in synaptic efficacy may underlie the acquisition of memory. Much interest has centred on the involvement of NMDA (N-methyl-D-aspartate) receptors in the induction of long-term potentiation (LTP), but since activation of this receptor alone results only in short-term enhancement, other receptors are clearly involved. This paper describes a novel form of lasting synaptic enhancement requiring concomitant activation of both NMDA and metabotropic quisqualate, or Qp receptors for induction. Neither high frequency presynaptic activity, co-activation of many presynaptic axons, postsynaptic depolarization, nor blockade of inhibition were required. However, repetitive, though relatively low frequency activity of the presynaptic axon appeared to be necessary. We suggest, tentatively, that presynaptic activity may be directly involved in priming the recently active terminal to respond to postsynaptic changes.     

Development of presynaptic inhibition onto retinal bipolar cell axon terminals is subclass-specific.

Synaptic integration is modulated by inhibition onto the dendrites of postsynaptic cells. However, presynaptic inhibition at axonal terminals also plays a critical role in the regulation of neurotransmission. In contrast to the development of inhibitory synapses onto dendrites, GABAergic/glycinergic synaptogenesis onto axon terminals has not been widely studied. Because retinal bipolar cells receive subclass-specific patterns of GABAergic and glycinergic presynaptic inhibition, they are a good model for studying the development of inhibition at axon terminals. Here, using whole cell recording methods and transgenic mice in which subclasses of retinal bipolar cells are labeled, we determined the temporal sequence and patterning of functional GABAergic and glycinergic input onto the major subclasses of bipolar cells. We found that the maturation of GABAergic and glycinergic synapses onto the axons of rod bipolar cells (RBCs), on-cone bipolar cells (ON-CBCs) and off-cone bipolar cells (OFF-CBCs) were temporally distinct: spontaneous chloride-mediated currents are present in RBCs earlier in development compared with ON- and OFF-CBC, and RBCs receive GABAergic and glycinergic input simultaneously, whereas in OFF-CBCs, glycinergic transmission emerges before GABAergic transmission. Because on-CBCs show little inhibitory activity, GABAergic and glycinergic events could not be pharmacologically distinguished for these bipolar cells. The balance of GABAergic and glycinergic input that is unique to RBCs and OFF-CBCs is established shortly after the onset of synapse formation and precedes visual experience. Our data suggest that presynaptic modulation of glutamate transmission from bipolar cells matures rapidly and is differentially coordinated for GABAergic and glycinergic synapses onto distinct bipolar cell subclasses.     

Temperature-sensitive conduction failure at axon branch points

1. The propagation of action potentials through the branching regions of squid axons was examined experimentally and with computer simulations over a temperature range of 5-25 degrees C. 2. Above a critical ratio of postbranch to prebranch diameters, propagation of an action potential failed. The value of this critical ratio is very sensitive to temperature and is smaller at high temperatures. The experimentally measured Q10 of the critical ratio is 0.37 +/- 0.04. 3. Evaluation of a number of parameters of action-potential propagation showed that this effect is closely related to the change in the width of the action potential with temperature (Q10 = 0.29 +/- 0.01).     

Synaptic transmission without action potentials: input-output properties of a nonspiking presynaptic neuron.

1. Input-output properties of the inhibitory synaptic connection between non-spiking neurons (EX1) and gastric mill (GM) neurons were examined in the stomatogastric ganglion of the spiny lobster, Panulirus interruptus. Current was injected into and the voltage was recorded during current injection, two independent microelectrodes were used. 2. The EX1-GM synaptic connection is a conductance-increase inhibitory type, with an input-output curve that resembles the curve for the squid giant synapse. There is a threshold level of depolarization for transmitter release from the presynaptic cell. Beyond that threshold, increasing presynaptic depolarization causes increasing postsynaptic hyperpolarization (and inhibition). 3. A long presynaptic current step always causes a postsynaptic response with an initial peak of hyperpolarization followed by a decay to a less hyperpolarized plateau level. The plateau level is maintained, in most cells, for the duration of the presynaptic depolarization even over long periods (30 s). 4. The peak, but not the plateau, part of the postsynaptic response is sensitive to the past history of the synaptic connection. If a large conditioning pulse is applied to the presynaptic cell causing a large postsynaptic hyperpolarization, then the postsynaptic response to a later presynaptic test depolarization will have a reduced peak, leaving the plateau component unchanged.     

Observations on the morphology of intracellularly stained gamma-motoneurons in relation to their axon conduction velocity.

Hindlimb gamma-motoneurons of adult cats were stained intracellularly with horseradish peroxidase. The gamma-motor-axons had intramedullary diameters between 2.0 micron and 2.4 micron and lacked recurrent collaterals. The conduction velocity of the gamma-motor-axons (20-29 m/sec) was close to what could be predicted from the relationship between conduction velocity and intramedullary diameter of much thicker adult alpha-motor-axons. However, the gamma-motor-axons were conducting much faster than alpha-motor-axons of 1-week-old kittens in spite of the fact that these two types of axons had about the same dimensions intramedullarly.     

Effects of peripheral axotomy on presynaptic axon terminals with GABA-like immunoreactivity

The facial nerve was unilaterally crushed at its exit from the stylomastoid foramen in three 3-month old male rats. After 10 days survival, before the regenerating axons had reinnervated their target muscles, the facial nucleus was examined to determine central patterns of response in material prepared to demonstrate the presence of GABA-like immunoreactivity with postembedding procedures using gold-labeled secondary antibody. The uninjured nucleus served as a control. In both control and injured nuclei, the GABAergic terminals synapse with all parts of the motor neurons, except the axon, and exhibit diverse morphologies. GABAergic axon terminals vary in their size and in the electron density of their axoplasm and the majority of the terminals contain pleomorphic vesicle profiles that display a range in their packing density and size. In both control and injured facial nuclei, only ～40% of the axon terminal profiles with pleomorphic vesicles exhibit GABA immunoreactivity. A morphometric analysis of the synaptic vesicle profiles in the GABA-positive terminals reveals that following axotomy there is no change in the mean number of synaptic vesicle profiles per GABAergic terminal profile. However, the mean size of the synaptic vesicle profiles in these terminals shows an axotomy-induced 50% increase, without change in the shapes of the enlarged vesicle profiles. Also, the numerical density of gold particles associated with the GABA-positive terminals is consistently greater in the injured than the control axon terminals. In the control animals quantitative analysis of the relative distribution of all axon terminal profiles in the neuropil categorized by the shape of their vesicle profiles as round, pleomorphic, or flat is 57:37:6. Ten days after axotomy the ratio of these categories in the injured nucleus has shifted to 35:60:5. This study demonstrates that the functional state of a postsynaptic target can influence the morphology of vesicle profiles in presynaptic elements as well as patterns of its afferent input. © 1994 Wiley-Liss, Inc.     

Synaptic targets of calretinin-containing axon terminals in macaque monkey prefrontal cortex

The coordinated activity of specific populations of pyramidal cells and GABA-containing, local circuit neurons in the primate prefrontal cortex (PFC) appears to be critical for working memory. Different subclasses of GABA-containing neurons can be distinguished by their content of the calcium-binding proteins parvalbumin (PV) and calretinin (CR). The postsynaptic targets of PV-containing cells have been well characterized in the primate PFC, but the postsynaptic targets of CR-containing neurons in this cortical region remain unknown. In the present study, we used immuno-electron microscopy to examine the synaptic type and postsynaptic targets of CR-immunoreactive (IR) axon terminals in the superficial and deep layers of macaque monkey PFC. Labeled axon terminals formed both symmetric and asymmetric synapses. Within the superficial layers, 93% of the synapses formed by CR-IR were symmetric, whereas in the deep layers the labeled axon terminals forming synapses were more evenly divided between symmetric (57%) and asymmetric (43%). The primary postsynaptic target of these two populations of CR-IR axon terminals also differed; unlabeled dendritic shafts were the predominant target of the symmetric synapses, whereas dendritic spines were the most common target of the asymmetric synapses. In addition, the mean cross-sectional area of the terminals forming asymmetric synapses was significantly larger than that of the terminals forming symmetric synapses. The presence of CR-IR asymmetric synapses suggested that they might arise from neurons that do not utilize GABA; indeed, dual-label fluorescent immunocytochemistry revealed that a subpopulation (23%) of CR-containing neurons in monkey PFC were not GABA-IR. These findings indicate that the synaptology of CR-containing neurons is more heterogeneous than that of PV-containing cells and suggests that the contributions of CR-containing neurons to cognitive processes mediated by the PFC may be more diverse.     

Identification of motor conduction block despite desynchronisation. A method

Identification of incomplete motor conduction block (CB) by the usual single stimulation method is problematic when impaired conduction velocity of the axons spared by the block exists at the site of the presumed CB, leading to desynchronisation. Indeed, the area of the compound muscle action potential (MAP) evoked by stimulation applied above this site may be reduced by the phase cancellation related to desynchronisation. A triple stimulation method with double collisions allows virtual displacement of the CB distally to the site where desynchronisation occurs. A first stimulus applied on the nerve above the site of the CB is followed--after a delay--by a stimulation applied distally on the nerve where a collision occurs. This collision, however, does not concern the antidromic wave of the blocked axons which travels towards the block. A third stimulus is then applied--after a second delay--below the site of the CB, where a second collision occurs in the blocked axons. Thus, the third stimulus evokes a MAP that concerns only the unblocked axons, as would the stimulus applied above the site of the CB. In this manner, desynchronisation occurring at the CB site is avoided. The method was developed on simulated CBs of the ulnar nerve at the elbow of 33 normal subjects and compared with the usual single stimulation method. Subsequently, it was evaluated on 8 pathological CBs of the ulnar nerve at the elbow and of the peroneal nerve at the fibular head. The method detects CBs of 5% or more despite the occurrence of desynchronisation.     

Spike timing-dependent long-term depression requires presynaptic NMDA receptors

NMDA receptors are necessary for both synaptic potentiation and depression, but the precise location of these receptors has not been established. By loading MK-801 into pre- or postsynaptic neurons during paired recordings of synaptically connected layer 4 and layer 2/3 neurons in mouse barrel cortex, we found that synaptic potentiation requires postsynaptic, but not presynaptic, NMDA receptors, whereas synaptic depression requires presynaptic, but not postsynaptic, NMDA receptors.     

Constancy of motor axon conduction time during growth in rats

Summary Axon conduction distance, conduction velocity, and conduction time were measured for individual triceps surae motoneurons in Sprague-Dawley rats weighing 230–630 g (i.e., age range 6–16 weeks). Both conduction distance (nerve length) and velocity were closely correlated with weight (r=0.95 and r=0.82, respectively). In contrast, conduction time did not change as weight increased nearly threefold. This striking constancy is probably due to a corresponding increase in axon diameter. It could contribute to maintenance of stable motor performance during rapid growth.     

Postsynaptic CPG15 promotes synaptic maturation and presynaptic axon arbor elaboration in vivo

The formation of CNS circuits is characterized by the coordinated development of neuronal structure and synaptic function. The activity-regulated candidate plasticity gene 15 (cpg15) encodes a glycosylphosphatidylinositol (GPI)-linked protein whose in vivo expression increases the dendritic arbor growth rate of Xenopus optic tectal cells. We now demonstrate that tectal cell expression of CPG15 significantly increases the elaboration of presynaptic retinal axons by decreasing rates of branch retractions. Whole-cell recordings from optic tectal neurons indicate that CPG15 expression promotes retinotectal synapse maturation by recruiting functional AMPA receptors to synapses. Expression of truncated CPG15, lacking its GPI anchor, does not promote axon arbor growth and blocks synaptic maturation. These results suggest that CPG15 coordinately increases the growth of pre- and postsynaptic structures and the number and strength of their synaptic contacts.     

Synaptic vesicle dynamics in the mossy fiber-CA3 presynaptic terminals of mouse hippocampus

The mossy fiber (MF)-CA3 synapse in the hippocampus is unique in the CNS because of its wide dynamic range of transmitter release during short- and long-term plasticity. The presynaptic mechanisms underlying the fidelity of transmission were investigated for the MF-CA3 synapses. The relative size of readily releasable pool (RRP) of vesicles was estimated by counting the number of docked vesicles at an active zone (AZ) on the transmission electron microscopy (TEM) image. The size of the releasable pool and the exo-endocytosis kinetics were directly measured from individual large MF boutons in hippocampal slices of transgenic mice that selectively express synaptopHluorin (SpH), a pH-sensitive GFP fused to the lumenal aspect of one of the vesicular membrane proteins, VAMP-2, in these boutons. Here we found (1) there are distinct two vesicle pools, the resting pool which is resistant to exocytosis, and the releasable pool, (2) the initially docked vesicles are easily depleted and the RRP is maintained by refilling from the reserve subpopulation of releasable pool (“reserve” releasable pool), and (3) the contribution of rapid reuse of recycled vesicles is relatively small. Therefore, the fidelity of transmission is suggested to be ensured by the rapid refilling rate of RRP.     

Synaptic vesicle dynamics in the mossy fiber-CA3 presynaptic terminals of mouse hippocampus

The mossy fiber (MF)-CA3 synapse in the hippocampus is unique in the CNS because of its wide dynamic range of transmitter release during short- and long-term plasticity. The presynaptic mechanisms underlying the fidelity of transmission were investigated for the MF-CA3 synapses. The relative size of readily releasable pool (RRP) of vesicles was estimated by counting the number of docked vesicles at an active zone (AZ) on the transmission electron microscopy (TEM) image. The size of the releasable pool and the exo-endocytosis kinetics were directly measured from individual large MF boutons in hippocampal slices of transgenic mice that selectively express synaptopHluorin (SpH), a pH-sensitive GFP fused to the lumenal aspect of one of the vesicular membrane proteins, VAMP-2, in these boutons. Here we found (1) there are distinct two vesicle pools, the resting pool which is resistant to exocytosis, and the releasable pool, (2) the initially docked vesicles are easily depleted and the RRP is maintained by refilling from the reserve subpopulation of releasable pool ("reserve" releasable pool), and (3) the contribution of rapid reuse of recycled vesicles is relatively small. Therefore, the fidelity of transmission is suggested to be ensured by the rapid refilling rate of RRP.