pH Dependence of transmission at electronic synapses of the crayfish septate axon

Gap junctions between segments of the crayfish septate axon mediate electronic transmission of impulses propagating along the length of the nerve cord. We simultaneously measured intracellular pH (pH_i) and gap junctional conductance (g_j) while axons were exposed to saline equilibrated with CO₂, weak acids, and the weak base ammonium chloride. Normal pH_i is about 7.1. When pH_i is elevated, g_j is unaffected. When pH_i is reduced, g_j declines with an apparent pK of about 6.7 and a Hill coefficient of about 2.7. We also measured effects of pH_i on non-junctional conductance (g_nj) and on the coupling coefficient, k. Over the pH_i range 6.2–8, g_nj increases approximately linearly with pH_i. Since k is a function of g_j and g_nj, it reached a maximum at about pH_i 7.1, decreasing at higher and lower pH_i. The pH_i dependence of g_j in crayfish septate axon is less steep and has a lower apparent pK than the g_j-pH_i relation in two vertebrate embryos previously examined. This finding illustrates a difference in gating among analoguos and possibly homologous membrane channels.     

Two classes of vesicles are present and change in relative proportion during post-embryonic development of rectifying electrical synapses in the crayfish.

The size and shape of vesicles at junctional appositions of the rectifying electrical synapses between the medial giant fibre and motor giant neurone of the crayfish were measured during the first 2 months after hatching. Summed data over this period reveal a bimodal distribution in vesicle diameter. From the day of hatching until about 7 days of age, small vesicles (circa 25 nm diameter) predominate. From day 7 onwards, larger vesicles (circa 55 nm diameter) occur in increasing numbers, until at day 56 they constitute about 85% of the population at any one junctional apposition. At intermediate ages (day 7-28) individual junctional appositions may show the same bimodal distribution in size as does the age group as a whole, indicating that large and small vesicles occur together at the same junction. The larger vesicles are mainly circular, while the small vesicles are pleomorphic, with shapes ranging from almost circular down to a shape factor of about 0.6.     

Renewal of electrotonic synapses in teleost retinal horizontal cells

In teleost retinas, the somata of same-type cone horizontal cells are electrically coupled via extensive gap junctions, as are the axon terminals of same-type cells. This coupling persists throughout the animal's life and is modulated by dopamine and conditions of light- vs. dark-adaptation. Gap junction particle density in goldfish horizontal cell somata has also been shown to change under these conditions, indicating that these junctions are dynamic. We have used electron microscopy to examine gap junctions in bass horizontal cells with a fixation method that facilitates detection of gap junctions. Annular gap junction profiles were observed in the somatic cytoplasm of all cone horizontal cell types in both light- and dark-adapted animals. Serial sections showed that most profiles represented gap junction vesicles free within the cytoplasm; the remainder represented vesicles still attached to extensive plasma membrane gap junctions by a thin cytoplasmic neck, suggestive of an intermediate stage in endocytosis. Observations of gap junction vesicles containing fragments of gap junctional membrane and/or fused with lysosomal bodies further supported this hypothesis. Because gap junctions persist between the horizontal cells, we propose that gap junction endocytosis and lysosomal degradation are balanced by addition of new junctions. While endocytosis has been widely demonstrated to serve in programmed removal of gap junctions (without subsequent replacement), from both nonneuronal cells and developing neurons, this study indicates that it can also function in the renewal of electrical synapses in the adult teleost retina, where gap junction elimination is not the goal.     

Development of habituation in the crayfish due to selective weakening of electrical synapses

During posthatching development, transmission becomes substantially reduced at monosynaptic electrical synapses between tactile afferents and the command neuron for caudal tailflip escape responses of the crayfish. The effectiveness of a parallel disynaptic pathway to the same command neuron is unaltered during posthatching growth. In small crayfish both the monosynaptic and disynaptic sensory pathways can elicit command cell action potentials. At this stage, the characteristics of the tailflip neural circuit are evidently controlled by the shorter latency, non-labile monosynaptic pathway. Consequently, tailflips can be reliably elicited in young crayfish, even at brief (2 s) interstimulus intervals. Tailflip responses of large, older crayfish are known to habituate when tactile stimuli are repeated at intervals up to 5 min. This decline in behavioral responsiveness is presumed to be mediated by low frequency synaptic depression (LFD) at first-order synapses of the disynaptic pathway. The lability of these synapses does not change during post-hatching development. Weakening of the monosynaptic pathway may be caused by command cell growth during posthatching development.     

Effects of levetiracetam on axon excitability and synaptic transmission at the crayfish neuromuscular junction

Levetiracetam (LEV) is a widely prescribed antiepileptic drug, but its actions on neuronal function are not fully characterized. Since this drug is believed to enter neurons by binding to a vesicular protein during endocytosis, we used motor axons of the crayfish opener neuromuscular junction to examine potential impacts of LEV on axon excitability. Two electrode current clamp from the inhibitory axon of the opener showed that LEV reduced action potential (AP) amplitude (AP_amp) and suppressed synaptic transmission, although the latter occurred with a longer delay than the reduction in AP_amp. Comparison of antidromic and orthodromic conducting APs in LEV suggested that this drug preferentially reduced excitability of the proximal axon, despite the expectation that it entered the axon at the terminals and should affect the distal branches first. Results presented here suggest that LEV modulates axonal excitability, which may in turn contribute to its antiepileptic effects.     

Null mutation in shaking-B eliminates electrical,but not chemical,synapses in the Drosophila giant fiber system: A structural study

Mutations in the Drosophila shaking-B gene perturb synaptic transmission and dye coupling in the giant fiber escape system. The GAL4 upstream activation sequence system was used to express a neuronal-synaptobrevin-green fluorescent protein (nsyb-GFP) construct in the giant fibers (GFs); nsyb-GFP was localized where the GFs contact the peripherally synapsing interneurons (PSIs) and the tergotrochanteral motorneurons (TTMns). Antibody to Shaking-B protein stained plaquelike structures in the same regions of the GFs, although not all plaques colocalized with nsyb-GFP. Electron microscopy showed that the GF-TTMn and GF-PSI contacts contained many chemical synaptic release sites. These sites were interposed with extensive regions of close membrane apposition (3.25 nm ± 0.12 separation), with faint cross striations and a single-layered array of 41-nm vesicles on the GF side of the apposition. These contacts appeared similar to rectifying electrical synapses in the crayfish and were eliminated in shaking-B² mutants. At mutant GF-TTMn and GF-PSI contacts, chemical synapses and small regions of close membrane apposition, more similar to vertebrate gap junctions, were not affected. Gap junctions with more vertebratelike separation of membranes (1.41 nm ± 0.08) were abundant between peripheral perineurial glial processes; these were unaffected in the mutants. J. Comp. Neurol. 404:449–458, 1999. © 1999 Wiley-Liss, Inc.     

Synaptic ultrastructure of Drosophila Johnston's organ axon terminals as revealed by an enhancer trap

The role of auditory circuitry is to decipher relevant information from acoustic signals. Acoustic parameters used by different insect species vary widely. All these auditory systems, however, share a common transducer: tympanal organs as well as the Drosophila flagellar ears use chordotonal organs as the auditory mechanoreceptors. We here describe the central neural projections of the Drosophila Johnston's organ (JO). These neurons, which represent the antennal auditory organ, terminate in the antennomechanosensory center. To ensure correct identification of these terminals we made use of a beta-galactosidase-expressing transgene that labels JO neurons specifically. Analysis of these projection pathways shows that parallel JO fibers display extensive contacts, including putative gap junctions. We find that the synaptic boutons show both chemical synaptic structures as well as putative gap junctions, indicating mixed synapses, and belong largely to the divergent type, with multiple small postsynaptic processes. The ultrastructure of JO fibers and synapses may indicate an ability to process temporally discretized acoustic information.     

Quantal release at visualized terminals of a crayfish motor axon: Intraterminal and regional differences

Synaptic transmission was measured at visualized terminal varicosities of the motor axon providing the sole excitatory innervation of the “opener” muscle in walking legs of crayfish (Procambarus clarkii Girard). Two questions were addressed: 1) How uniform is quantal emission at different locations along terminals innervating a single muscle fiber, and 2) can differences in quantal emission account for the different excitatory postsynaptic potential (EPSP) amplitudes generated by terminals localized in defined regions of the muscle? Extracellular “macropatch” electrodes were placed over individual varicosities, viewed after brief exposure to a fluorescent dye, and synaptic currents were recorded to determine quantal content of transmission. Along terminals supplying a single muscle fiber, nonuniform release was found: Varicosities closer to the point of origin of the terminal branch released more transmitter than those located more distally. Quantal content was higher for varicosities of the muscle's proximal region (where large EPSPs occur) than for varicosities of the central region (where small EPSPs occur). The probability of transmitter release per synapse is estimated to be greater for the proximal varicosities. At low frequencies of stimulation, quantal content per muscle fiber is two to four times larger in the proximal region. Taken in conjunction with a twofold higher mean input resistance for the proximal muscle fibers, the difference in quantal content can account for a four- to eightfold difference in EPSP amplitude. The observed mean EPSP amplitude is at least eight times larger in the proximal region. We discuss factors contributing to differences in EPSP amplitudes. © 1996 Wiley-Liss, Inc.     

Ultrastructure of the electrotonic and chemical components of the lateral-to-motor and medial-to-motor synapses in crayfish nerve cord

Lateral-to-motor and medial-to-motor synapses in crayfish nerve cords are composed of an electrical and a chemical component. The presynaptic terminals showed localized clusters of synaptic vesicles, electron-dense areas, coated pits, and coated vesicles. In thin sections, active zones were defined by electron-dense regions where synaptic vesicles attached and, in freeze-fracture replicas, by clusters of intramembrane particles localized in bands with vesicle openings on the sides of these bands. The cytoplasmic surface of the postsynaptic membrane opposite the active zones was coated with electron-dense material that in freeze-fracture replicas was seen as an increase in intramembrane particles located in the external leaflet (EF-face). This specialization of the postsynaptic membrane may correspond to the neurochemical receptor. Also, pre- and postsynaptic membranes were separated by a wider extracellular gap than those of adjacent nonsynaptic regions and electrical synapses or gap junctions. Synaptic vesicles were located exclusively at the synaptic regions by means of a cytoskeleton that was different for the electrical and the chemical components. The vesicles associated with the electrical component were anchored to a cytoskeleton composed of a beaded layer of densities located parallel to the membrane. This cytoskeleton maintained the synaptic vesicles separated from the presynaptic membrane by a distance of 13 +/- 2 nm. The synaptic vesicles associated with the chemical component were anchored to electron-dense regions formed by filaments arranged in bundles, anchored to the presynaptic membrane. Vesicles lined both sides positioned to discharge their contents into the extracellular space and to replace the discharged vesicles.     

Freeze-fracture study of the large myelinated club ending synapse on the goldfish Mauthner cell: special reference to the quantitative analysis of gap junctions

The large myelinated club endings (LMCEs) of primary eighth nerve afferents form mixed synapses on the lateral dendrite of the giant Mauthner cell. The double replica freeze-fracture technique was employed to examine the intramembrane fine structure of these LMCE synapses. Morphological correlates of both chemical and electrical transmission were found at the LMCE synapses. Electrical synaptic junctions, or gap junctions, were located over much (10-20%) of the synaptic contact. These were seen in both pre-and postsynaptic membrane as tightly packed P face particle aggregates and corresponding aggregates of E face pits. Specializations characteristic of chemical synaptic junctions were most prominent at the periphery of the synaptic contact. These specializations consisted of postsynaptic E face particle aggregates which were subjacent to presynaptic active zones. The active zones were distinguishable as regions with an increased density of large particles and vesicle attachment sites represented by P face depressions and E face protuberances. Quantitative analysis of gap junction particle (connexon) number at five LMCEs revealed 24,000-106,000 connexons per LMCE. Comparison with data from electrophysiological studies of single LMCEs indicates that only a small fraction of the connexon channels are open at any given time during electrotonic transmission at an LMCE synapse.     

Axotomy-induced temporal dissociation of long-term adaptive changes at neuromuscular synapses of a crayfish phasic motoneuron

Periodic in situ stimulation of an identified crayfish phasic extensor motoneuron for 3 consecutive days (2 h/day) at 2.5 Hz leads to long-term adaptation (LTA) of its neuromuscular synapses. LTA is characterized by reductions in both initial excitatory postsynaptic potential (EPSP) amplitudes and synaptic depression during repeated stimulation. These adaptive changes were evident 1 day following periodic stimulation. Axotomy of the motoneuron before or after the first day of stimulation of its distal surviving axon abolished both adaptive changes. Axotomy between the second and third stimulation periods abolished only the resistance to synaptic depression. Both adaptive changes were expressed following axotomy after the third day of stimulation. Axotomy alone did not affect neuromuscular transmission in control, unstimulated animals. These results show that axonal continuity between the phasic extensor motoneuron's cell body and its neuromuscular synapses is required at specific times during periodic stimulation for the expression of each of these long-term adaptive changes in neuromuscular transmission. Furthermore, the two adaptive changes in transmission are temporally separable, with the resistance to depression requiring more periodic stimulation to emerge than the reduction in initial EPSP amplitudes. The results also suggest that the molecular components responsible for the expression of these adaptive changes are synthesized in the soma and transported down the axon in response to periodic stimulation of the phasic axon.     

TwoB or not twoB: differential transmission at glutamatergic mossy fiber-interneuron synapses in the hippocampus

Mossy fiber (MF) synapses are key stations for flow of information through the hippocampal formation. A major component of the output of the MF system is directed towards inhibitory interneurons. Recent studies have revealed that the functional properties of MF–interneuron synapses differ substantially from those of MF–CA3 pyramidal neuron synapses. Mossy-fiber–interneuron synapses in the stratum lucidum represent a continuum of functional subtypes, in which the subunit composition of postsynaptic AMPA receptors and NMDA receptors appears to be regulated in a coordinated manner.     

Development of the rhesus monkey retina. I. Emergence of the inner plexiform layer and its synapses

The emergence and differentiation of the inner plexiform layer (IPL) and the establishment of its synapses were analyzed in retinae of 18 rhesus monkeys ranging in age from the 55th embryonic day (E55) to 10 years. The IPL becomes recognizable by E65 as a thin acellular zone consisting of immature neurites and growth cones scattered within large extracellular spaces. In each specimen, apposing paired membrane specializations were classified as junctions without synaptic vesicles, conventional synapses, ribbon synapses, or gap junctions. Initially, at E65, the IPL consists of variety of immature cell processes that are interconnected exclusively by junctions without synaptic vesicles, at a density of 4.7/100 microns2. By E73, the IPL becomes more distinct and wider and contains 7.8 such junctions/100 microns2. Conventional synapses develop by the addition of vesicles to initially vesicle-free junctions. The first conventional synapses appear at E78. They increase in density from 1.5 to 3.2/100 microns2 between E78 and E84 and reach a density of 7.9/100 microns2 by E99. A rapid burst in synaptogenesis occurs in the IPL between E99 and E114; a density of 16.5/100 microns2 is reached, mainly due to accretion of conventional synapses. Ribbon synapses first become recognizable at E99, almost 3 weeks after the emergence of conventional synapses. By E114 they account for about 7% of all synaptic contacts in the IPL. The rate of synaptogenesis slows down during the last quarter of gestation; the adult level of about 24 contacts/100 microns2 is reached between E130 and E149. Of these, 72.2% are of conventional type, 15.4% are ribbon synapses, and 12.4% are junctions without vesicles. However, in the adult the density of junction without vesicles is only about one-half that found at E149. Gap junctions are absent during the initial and rapid phases of synaptogenesis; they appear abruptly, between E130 and E149, only after the density of chemical synapses in the IPL has reached the adult level. In the rhesus monkey, synaptogenesis begins several weeks later in the IPL than in its primary targets--the dorsal lateral geniculate nucleus and the superior colliculus (Hendrickson and Rakic, '77; Cooper and Rakic, '83). However, the rapid increase in density of conventional synapses in the IPL coincides with the segregation of retinal projections from right and left eyes in the geniculate nucleus (Rakic, '76) and with the elimination of the large surplus of retinal ganglion cell axons (Rakic and Reley, '83).     

Localization of glutamic-acid-decarboxylase-immunoreactive axon terminals in the inferior olive of the rat, with special emphasis on anatomical relations between GABAergic synapses and dendrodendritic gap junctions

Immunocytochemical and electron microscopic methods were used to examine the GABAergic innervation of the inferior olivary nucleus in adult rats. This neuronal system was visualized with an antibody against glutamic acid decarboxylase (GAD, EC 4.1.1.15), the GABA-synthesizing enzyme. A GAD-positive reaction product was encountered only in short segments of preterminal axons and in axon terminals. Their relative number per unit area of neuropil was very similar in all olivary subnuclei. Despite this homogeneity in density, obvious intraregional differences existed. Some regions were strongly immunoreactive (the "c" subgroup, the beta nucleus, and the mediolateral outgrowth of the medial accessory olive), whereas others were weakly labeled (the dorsomedial cell column and the central zones of the medial accessory and principal olives). The strongly immunoreactive areas contained the largest and most intensively labeled axon terminals. Areas of weak labeling were filled with small, weakly immunoreactive nerve terminals. Thus, variations in size and in intensity of labeling create a specific pattern of GABA innervation, revealed by an almost continuous gradient between the above-mentioned extremes. The GAD-positive axon terminals established conventional synapses with dendrites (94% of the samples) or with cell bodies (6%). The vast majority of these synapses were type II (84%) and only a small proportion formed type I synaptic contacts (16%), regardless of the nature of the postsynaptic element. Immunoreactive terminals were also involved in the complex synaptic arrangements--the glomeruli, which characterize the olivary neuropil. Within these formations, olivary neurons were electrotonically coupled through dendrodendritic gap junctions. There was a constant association between GAD-positive axon terminals and small dendritic appendages linked by gap junctions. This association was revealed not only by the systematic presence of immunolabeled terminals directly apposed to the dendritic appendages but, more importantly, by the frequent presence of type II synapses straddling both elements. These synapses were in close proximity to the low-resistance pathways represented by the gap junctions. The strategic location of these GABA synapses is discussed in relation to recent findings indicating the possibility of a synaptic modulation of the electrical coupling: the release of GABA, by increasing nonjunctional membrane conductance, could shunt the coupling between olivary neurons. The functional decoupling of selected gap junctions would be responsible for the spatial organization of the olivary electrotonic coupling.     

Morphological and functional study of dwarf neurons in the rat striatum

Combination of morphological and electrophysiological techniques provided data, suggesting existence in the young rat striatum of a peculiar class of neurons, the neurogliaform or dwarf neurons. Striatal neurons (n = 92), intracellularly recorded from rat brain slices, were filled (one in each slice) with the intracellular marker biocytin, to compare physiological and morphological properties in the same cell. Moreover, some neurons (n = 7) were filled with biocytin plus the fluorescent calcium indicator fura-2, identifying cells during electrophysiological recording. Electrophysiological recordings showed that striatal neurons had different firing patterns, suggestive in most cases (n = 80) of spiny neuron class and in others (n = 12) of interneuron class. Fura-2 injection clearly identified the body of six medium-sized cells and of one distinctive tiny cell. This small cell, however, showed a resting membrane potential and spontaneous and evoked firing pattern characteristic of striatal interneurons. Moreover, the fura-2 injected in such small neuron also completely filled the cell body of a near large neuron; the fura-2 fluorescence changed synchronously in the two paired neurons after electrical stimulation of the impaled small one. Accordingly, the biocytin staining identified the morphology of the small recorded neuron as a neurogliaform-like cell apposed to a dendrite of an aspiny neuron, suggesting that the dye injected in one neuron had diffused to the other of a different type. Furthermore, such heterologous dye coupling unexpectedly involved seven pairs of cells detected with biocytin staining (7.6% of the recorded neurons), invariably represented by a medium or large neuron on one side, and on the other side by a small (5.44 ± 0.15 × 9.14 ± 0.7 μm, mean ± SD;n = 7) neurogliaform cell, roundish in shape with few slender and short processes, usually apposed to a dendrite of the companion neurons (six out of seven). In the other cases, the biocytin staining revealed in each slice either the morphology of single spiny or aspiny neurons (80.4% of recorded neurons), or of two–three medium-sized spiny neurons detected near to each other, suggesting that dye coupling had occurred typically between similar neurons (11.9% of the recorded neurons). These data suggest that some neurogliaform cells in the striatum of young rat can be identified as dwarf interneurons, that may be dye-coupled with neurons of different classes.     

Autoregulatory properties of dorsal raphe 5-HT neurons: Possible role of electrotonic coupling and 5-HT1D receptors in the rat brain

In the present study, the hypothesis that somatodendritic availability of 5-hydroxytryptamine (5-HT) could be regulated independently of the firing activity of dorsal raphe 5-HT neurons was tested. The 5-HT pathway was electrically stimulated at the level of the ventromedial tegmentum and the ensuing action potentials, recorded in the dorsal raphe, met all criteria for antidromic invasion of 5-HT neurons. The latency of antidromic spikes was current-dependent and the changes in latency were of quantal nature. This observation suggests an electrotonic coupling between 5-HT neurons. Stimulation of the ventromedial tegmentum also induced a decrease in the probability of firing of 5-HT neurons. This reduction in 5-HT neuron firing activity is a 5-HT-mediated response, due to an increased bioavailability of the neurotransmitter in the biophase of somatodendritic 5-HT_1A autoreceptors. The intravenous administration of the 5-HT₁ agonists TFMPP and RU 24969 reduced the duration of suppression of firing induced by the 5-HT-pathway stimulation, without altering the spontaneous firing rate of 5-HT neurons. The effect of TFMPP and RU 24969 on duration of suppression was blocked by (±)mianserin, a drug with high affinity for the rat 5-HT_1D, but not 5-HT_1B, receptors. On the other hand, (−)propranolol, a mixed 5-HT antagonist also blocked the effect of TFMPP. However, the selective 5-HT_1A antagonist (+)WAY 100135 did not alter the effect of TFMPP. These results, in keeping with previous anatomical studies, suggest the existence of electrotonic coupling of 5-HT neurons and indicate that 5-HT release in the rat dorsal raphe nucleus may be controlled independently of firing-regulating 5-HT_1A autoreceptors. They also suggest that 5-HT_1D receptors may play a role in this regulatory function of 5-HT neurons. © 1996 Wiley-Liss, Inc.     

Localization of heterotypic gap junctions composed of connexin45 and connexin36 in the rod pathway of the mouse retina

The primary rod pathway in mammals contains gap junctions between AII amacrine cells and ON cone bipolar cells which relay the rod signal into the cone pathway under scotopic conditions. Two gap junctional proteins, connexin36 (Cx36) and connexin45 (Cx45), appear to play a pivotal role in this pathway because lack of either protein leads to an impairment of visual transmission under scotopic conditions. To investigate whether these connexins form heterotypic gap junctions between ON cone bipolar and AII amacrine cells, we used newly developed Cx45 antibodies and studied the cellular and subcellular distribution of this protein in the mouse retina. Specificity of the Cx45 antibodies was determined, among others, by Western blot and immunostaining of mouse heart, where Cx45 is abundantly expressed. In mouse retina, Cx45 immunosignals were detected in both plexiform layers and the ganglion cell layer. Double staining for Cx45 and Cx36 revealed a partial overlap in the punctate patterns in the ON sublamina of the inner plexiform layer of the retina. We quantified the distributions of these two connexins in the ON sublamina, and detected 30% of the Cx45 signals to be co-localized with or in close apposition to Cx36 signals. Combining immunostaining and intracellular dye injection revealed an overlap or tight association of Cx36 and Cx45 signals on the terminals of injected AII amacrine and two types of ON cone bipolar cells. Our results provide direct evidence for heterotypic gap junctions composed of Cx36 and Cx45 between AII amacrine and certain types of ON cone bipolar cells.     

Gap junctions among the perikarya, dendrites, and axon terminals of the luminosity-type horizontal cell of the turtle retina

Gap junctions of the H1 horizontal cell of the turtle retina (Leeper, '78) were studied in thin-sectioned material and in freeze-fracture replicas. Perikaryal gap junctions were extremely restricted, 0.02-0.07 micron2 in in area, whereas those of axon terminals were much larger, most being 0.1-1.0 micron2. Both varieties, however, had the usual seven-layered appearance in thin section and measured 15 +/- 1 nm in overall width between cytoplasmic faces. Freeze-fractured views of the perikaryal junctions revealed roughly circular patches of P-face 9-nm particles and E-face pits. The axon terminal gap junctions were seen as large areas of P-face particles and E-face pits containing occasional islands of unspecialized membrane. Particle densities varied from 1,455 to 2,448 microns-2. A serial reconstruction was made of a portion of the axon terminal network in order to measure the surface areas of the axons contained therein and the fraction occupied by gap junctions. These data demonstrated that the fractional area occupied by gap junctions was roughly in inverse proportion to the area of the axon region (tuberous core vs. terminal process). It is argued that this constitutes an impedance matching device to ensure adequate current flow through the axon processes. Assuming that each P-face particle represents a connection having a conductance of 10(-10) S and given the P-face particle density and gap junctional areas determined in this report, we calculated that the gap junction distribution is adequate to account for the spatial properties of the horizontal cell axon network (Lamb, '76).     

Involvement of brain‐derived neurotrophic factor (BDNF) in the functional elimination of synaptic contacts at polyinnervated neuromuscular synapses during development

We use immunohistochemistry to describe the localization of brain‐derived neurotrophic factor (BDNF) and its receptors trkB and p75^NTR in the neuromuscular synapses of postnatal rats (P6–P7) during the synapse elimination period. The receptor protein p75^NTR is present in the nerve terminal, muscle cell and glial Schwann cell whereas BDNF and trkB proteins can be detected mainly in the pre‐ and postsynaptic elements. Exogenously applied BDNF (10 nM for 3 hr or 50 nM for 1 hr) increases ACh release from singly and dually innervated synapses. This effect may be specific for BDNF because the neurotrophin NT‐4 (2–8 nM) does not modulate release at P6–P7. Blocking the receptors trkB and p75^NTR (with K‐252a and anti‐p75‐192‐IgG, respectively) completely abolishes the potentiating effect of exogenous BDNF. In addition, exogenous BDNF transiently recruits functionally depressed silent terminals, and this effect seems to be mediated by trkB. Calcium ions, the L‐type voltage‐dependent calcium channels and protein kinase C are involved in BDNF‐mediated nerve ending recruitment. Blocking experiments suggest that endogenous BDNF could operate through p75^NTR receptors coupled to potentiate ACh release in all nerve terminals because the anti‐p75‐192‐IgG reduces release. However, blocking the trkB receptor (K‐252a) or neutralizing endogenous BDNF with the trkB‐IgG fusion protein reveals a trkB‐mediated release inhibition on almost mature strong endings in dual junctions. Taken together these results suggest that a BDNF‐induced p75^NTR‐mediated ACh release potentiating mechanism and a BDNF‐induced trkB‐mediated release inhibitory mechanism may contribute to developmental synapse disconnection. © 2009 Wiley‐Liss, Inc.