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Y Ben-Ari

Publications and source records attributed to Y Ben-Ari.

At least 145 records · Page 8Linked to original sources
PubMed1995

Gliosis and axonal sprouting in the hippocampus of epileptic rats are associated with an increase of tenascin-C immunoreactivity.

Temporal lobe epilepsy is associated with neuronal death, gliosis and sprouting of mossy fibres in the hippocampus of human and rats. In the present study we show that immunoreactivity for tenascin-C (an extracellular matrix glycoprotein) increase in the hippocampus of epileptic rats. However, this increase was only observed in the cases displaying neuronal cell loss and glial reaction (i.e. after kainate treatment but not after kindling). Tenascin-C increase was particularly striking at Ammon's horn, where the antibody labelled both reactive astrocytes (confirmed by double-labelling experiments) and axonal plasma membranes. In the molecular layer tenascin-C immunoreactivity remained unchanged in both kindled or kainate treated rats. It is interesting that increased tenascin-C immunoreactivity was observed within zones in which axonal regeneration did not occur (the CA3 area in kainate-treated animals) whereas zones in which reactive synaptogenesis occurred (such as the CA3 area of kindled rats or the molecular layer of both kindled and kainate-treated rats) were devoid of tenascin-C immunoreactivity. We infer from these results that tenascin-C impedes the terminal sprouting of mossy fibres in CA3 of kainate-treated rats.

Animals↗
PubMed1995

NMDA receptor redox sites: are they targets for selective neuronal protection?

NMDA receptors play a central role in neuronal plasticity and in several pathological situations. Transient activation of this receptor triggers long-term potentiation, whereas sustained activation leads to cell death. Evidence for control of this activity by a redox site in cell cultures, brain tissues and in recombinant NMDA receptors are discussed by Henri Gozlan and Yehezkel Ben-Ari. The characteristics of this modulation and the consequences of redox state modifications on NMDA-mediated events are examined in vitro under physiological and pathological conditions. Since metabolic disorders enhance NMDA receptor function, the redox site could constitute a new target for selectively preventing in vivo the deleterious consequences of overactivation without blocking neuronal plasticity mediated by NMDA receptors.

Alkylation↗
PubMed1995

Early endonuclease activation following reversible focal ischemia in the rat brain.

The structural changes that occur in chromatin DNA after ischemic brain injury are poorly understood. The presence of oligonucleosome fragments that are recognized as the characteristic DNA ladder has been demonstrated in global and focal ischemia, associated or not with random DNA fragmentation. Using pulsed-field gel electrophoresis, which improves DNA separation, we have now detected initial stages of DNA fragmentation that occur already 6 h after reversible focal cerebral ischemia in rats. This result confirms that internucleosomal DNA fragmentation precedes random DNA fragmentation in vulnerable striatal and cortical neurons following reversible focal cerebral ischemia.

Animals↗
PubMed1995

Cytosine arabinoside induces apoptosis in cerebellar neurons in culture.

Cytosine arabinoside (AraC) is a pyrimidine antimetabolite that prevents cell proliferation by inhibiting DNA synthesis. We report that AraC kills cultured cerebellar neurons in a concentration-dependent fashion with an EC50 of approximately 60 microM when added shortly after seeding. This cell death has apoptotic features because we observed (1) morphology of apoptotic nuclei as judged by DNA staining with Hoechst 33258, (2) DNA fragmentation with typical ladder pattern on agarose gel, (3) positive nuclear labeling with a specific in situ DNA fragmentation staining, (4) prevention by deoxycytidine (IC50 = 1 microM), protein, and RNA synthesis inhibitors, and (5) release of DNA fragments in the incubating medium. We have also observed that several proteins were overexpressed in AraC-treated neurons by two-dimensional polyacrylamide gel electrophoresis. We conclude that AraC induces a signal that triggers a cascade of new mRNA and protein synthesis, leading to apoptotic cell death in cultured cerebellar granule cells.

Animals↗
PubMed1995

Different GABAB-mediated effects on protein kinase C activity and immunoreactivity in neonatal and adult rat hippocampal slices.

The effects of GABA on protein kinase C (PKC) were investigated in rat hippocampal slices at various postnatal ages [postnatal day (P) 1-P60]. At P4, GABA (300 microM) induced a rapid (in 1-2 min) 40-50% increase of PKC activity in the membrane fraction and a decrease in the cytosol. These effects were mediated by GABAB receptors because (a) they were neither blocked by 10 microM bicuculline nor reproduced by 10 microM isoguvacine and (b) they were mimicked by the GABAB agonist baclofen (3-30 microM), an effect fully antagonized by the GABAB antagonist 2-hydroxysaclofen (10 microM). A baclofen-induced increased PKC activity in the membrane fraction was only present during the early postnatal period (P1-P14); it was associated with a translocation from the cytosol to the membrane of the immunoreactivity of some PKC isoforms (alpha-, beta-, and epsilon-PKCs). In contrast, after P21, PKC activity and alpha-, beta-, epsilon-, and gamma-PKC immunoreactivities were decreased by baclofen in the membrane fraction and increased in the cytosol. These results suggest that the stimulation of GABAB receptors differentially modulates PKC activity via distinct second messenger pathways in developing and mature hippocampi.

Aging↗
PubMed1995

Postnatal development of pre- and postsynaptic GABAB-mediated inhibitions in the CA3 hippocampal region of the rat.

1. Intracellular recordings were made from adult and neonatal rat hippocampal slices to study the postnatal development of GABAB-mediated inhibition in CA3 pyramidal neurons. 2. In the presence of glutamatergic receptor antagonists, direct electrical stimulation of the interneurons induced a biphasic GABAA- and GABAB-mediated inhibitory postsynaptic potential in adult [postnatal day (P) 30-P40] and young (P6-P8) CA3 pyramidal neurons. In contrast, in pups (P0-P3), electrical stimulation only induced a bicuculline-sensitive depolarizing GABAA synaptic potential. 3. The outward postsynaptic currents generated by bath-applications of baclofen (30 microM, 30 s) at P3 (78 +/- 60 pA, mean +/- SE) were 4 to 5 times smaller than those evoked between P6 (329 +/- 32 pA) and P30 (412 +/- 44 pA). At P0, baclofen failed to induce a postsynaptic current. 4. The outward currents generated by serotonin (50 microM, 30 s) and the A1 receptor agonist N-cyclopentyladenosine (40 microM, 30 s) ranged between 0 and 50 pA at P3 and between 200 and 400 pA at P6 and P30 (holding potential = -60 +/- 2 mV). 5. In the presence of potassium channel blockers, the amplitude of calcium current elicited by a depolarizing voltage step command (1 s) from a holding potential of -60 mV to a test potential of 0 mV was 2 +/- 0.15 nA at P6 (n = 9) and 0.73 +/- 0.14 nA at P3 (n = 8). Baclofen reversibly reduced the amplitude of calcium currents in young rats but not in pups. 6. Baclofen reversibly reduced the amplitude of the evoked GABAA-mediated and glutamatergic synaptic events at all developmental stages. These effects were dose dependent and antagonized by P-alpha 3-aminopropyl-P-diethoxymethyl-phosphinic acid (CGP) 35348 (500 microM). 7. We conclude that postsynaptic GABAB-mediated inhibition is absent or minimal during the first postnatal days in the CA3 region. In contrast, presynaptic GABAB inhibition is present at birth. We discuss the mechanisms and physiological consequences of these observations.

6-Cyano-7-nitroquinoxaline-2,3-dione↗
PubMed1995

Direct demonstration of functional disconnection by anoxia of inhibitory interneurons from excitatory inputs in rat hippocampus.

1. We studied the effects of anoxia on excitatory and inhibitory postsynaptic currents (EPSCs and IPSCs) evoked by electrical stimulation in the stratum radiatum in concomitantly recorded pyramidal cells and interneurons of the CA1 region of rat hippocampal slices. We used the blind whole cell patch-clamp technique, and anoxia was induced by switching perfusion of the slice from oxygenated artificial cerebral spinal fluid (ACSF) to ACSF saturated with 95% N2-5% CO2 for 4-6 min. 2. As in pyramidal neurons, anoxia induced in interneurons outward currents, during and shortly after the anoxic episode. Both currents were, however, significantly larger in interneurons than in pyramidal neurons. 3. EPSCs are more rapidly depressed by anoxia in interneurons than in simultaneously recorded pyramidal cells. 4. In pyramidal neurons, polysynaptic IPSCs (pIPSCs) evoked by conventional distant stimulation (> 1 mm) are more sensitive to anoxia then EPSCs. In contrast, in interneurons, anoxia blocks with a similar latency EPSCs and polysynaptic IPSCs. 5. To determine whether this block of pIPSCs in pyramidal cells is due to a shift in driving force or a change in conductance, we examined the current (I/V) relationships. The block by anoxia of pIPSCs is due to a reduction of IPSC conductance (> 98%) that occlude other events including the shift of IPSCs reversal potential (ECl).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

In CA1 hippocampal neurons, the redox state of NMDA receptors determines LTP expressed by NMDA but not by AMPA receptors.

1. Using extracellular recording techniques in the CA1 region of the rat hippocampus, we have evaluated the effects of the redox reagents 5,5O-dithiobis-2-nitrobenzoic acid (DTNB) and tris (carboxyethyl) phosphine (TCEP) on long-term potentiation (LTP) expressed by alpha-amino-3-hydroxy-5-methyl-isoxazole-4-propionic acid (AMPA) and N-methyl-D-aspartate (NMDA) receptors. In physiological conditions a high-frequency stimulation (HFS) of Schaffer collateral-commissural fibers induced a LTP expressed by a persistent increase (73 +/- 13%, mean +/- SE, n = 8/10) of AMPA field potentials (LTPA). In the presence of 10 microM of 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) and reduced concentration of Mg2+ (0.1 mM) to boost NMDA receptors, the HFS induced LTP of NMDA field potentials (LTPN; 62 +/- 11%, n = 8/10). 2. The thiol-oxidizing reagent DTNB (200 microM) reduced, by 46 +/- 5% (n = 24), NMDA-receptor field potentials (NMDA-FP), and this effect could not be reversed by extensive washing. The disulfide-reducing agent TCEP (200 microM) slightly increased AMPA-FP and reversed the DTNB-induced inhibition of NMDA-FP. 3. DTNB (200 microM, 10 min), and TCEP (200 microM, 20 min), had no effect on AMPA-FP (98 +/- 3% and 101 +/- 5%, respectively, n = 12). 4. DTNB (200 microM, 15 min) did not prevent the induction or expression of LTPA (-12 and -5%, respectively, n = 8/8). Similar results were observed with TCEP (200 microM, 20 min).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

Hippocampal CA1 lacunosum-moleculare interneurons: modulation of monosynaptic GABAergic IPSCs by presynaptic GABAB receptors.

1. Whole cell patch-clamp recordings were employed to characterize monosynaptic inhibitory postsynaptic currents (IPSCs) in morphologically and electrophysiologically identified interneurons located in the stratum lacunosum moleculare, or near the border of the stratum radiatum (LM interneurons), in the CA1 region of hippocampal slices taken from 3- to 4-wk-old rats. Monosynaptic IPSCs, evoked in the presence of glutamate receptor antagonists 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX; 20 microM) and D-2-amino-5-phosphopentanoate (APV; 50 microM) were biphasic. The gamma-aminobutyric acid-A (GABAA) receptor antagonist, bicuculline (20 microM), blocked the fast IPSC, and the slow IPSC was blocked by the GABAB receptor antagonist CGP35348 (500 microM). 2. Monosynaptic IPSCs were evoked by electrical stimulation in several distant regions including the stratum radiatum, the stratum oriens, the stratum lacunosum-moleculare, and the molecular layer of dentate gyrus, suggesting an extensive network of inhibitory interneurons in the hippocampus. In paired recordings of CA1 interneurons and pyramidal cells, IPSCs were evoked by electrical stimulation of most of these distal regions with the exception of the molecular layer of dentate gyrus, which evoked an IPSC only in LM interneurons. 3. Frequent (> 0.1 Hz) stimulation depressed the evoked IPSCs. With a paired-pulse protocol, the second IPSC was depressed and the maximal depression (40-50%) was observed with an interstimulus interval of 100-200 ms. 4. The GABAB receptor agonist baclofen (1 microM) reduced the amplitude of evoked IPSCs and the paired-pulse depression of the second IPSC. The GABAB receptor antagonist CGP35348 (0.5-1 mM) had no significant effect on the amplitude of isolated IPSCs. However, CGP35348 reduced but did not fully block paired-pulse depression, suggesting that this depression is partly due to the activation of presynaptic GABAB receptors. 5. The paired-pulse depression depended on the level of transmitter release. Potentiation of synaptic release of GABA, by increasing the extracellular Ca2+ concentration to 4 mM and reducing the extracellular Mg2+ concentration to 0.1 mM, enhanced the depression. Reduction of transmitter release by increasing extracellular Mg2+ concentration to 7 mM diminished the paired-pulse depression of IPSCs. After potentiation of transmitter release, CGP35348 was less efficient in reducing the paired-pulse depression, suggesting that enhancement of depression by high-calcium/low-magnesium medium was preferentially due to the potentiation of a GABAB-independent component. 6. In summary, monosynaptic IPSCs recorded in LM interneurons show similar features to those recorded in pyramidal cells. The strong correlation between the level of transmitter release and the degree of paired-pulse depression may have important physiological consequences, because in synapses with a high level of activity and a high level of GABA release, inhibition is powerful, but depression can develop more readily.

6-Cyano-7-nitroquinoxaline-2,3-dione↗
PubMed1995

Hippocampal CA1 lacunosum-moleculare interneurons: comparison of effects of anoxia on excitatory and inhibitory postsynaptic currents.

1. The effects of anoxia on excitatory and inhibitory postsynaptic currents (EPSCs and IPSCs, respectively) evoked by electrical stimulation in the stratum radiatum were studied in morphologically and electrophysiologicaly identified lacunosum-moleculare (LM) interneurons of the CA1 region of rat hippocampal slices. The blind whole cell patch-clamp technique was used, and anoxia was induced by superfusion of the slice with an anoxic artificial cerebral spinal fluid saturated with 95% N2-5% CO2 for 4-6 min. 2. In LM interneurons, anoxia generated currents similar to those in pyramidal cells, the most prominent being anoxic and postanoxic outward currents. The adenosine A1 type receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX, 200 nM) did not significantly affect anoxia-generated currents. 3. EPSCs and polysynaptic IPSCs (pIPSCs) evoked in LM interneurons by "distant" stimulation (> 1 mm) in the stratum radiatum were strongly depressed by anoxia and recovered upon reoxygenation. 4. Responses to pressure application of glutamate, alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA), and N-methyl-D-aspartate (NMDA) were not significantly affected by anoxia, suggesting that the suppression of EPSCs is due to presynaptic mechanisms. 5. DPCPX (200 nM) prevented anoxia-induced suppression of EPSCs, suggesting that this suppression was mediated by presynaptic A1 adenosine receptors. 6. Monosynaptic IPSCs evoked by "close" stimulation (< 0.5 mm) in the stratum radiatum, in the presence of glutamate-receptor antagonists 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX; 20 microM) and D-2-amino-5-phosphopentanoate (APV; 50 microM), were reversibly depressed but not blocked by anoxia. 7. Anoxia depressed monosynaptic GABAA receptor-mediated IPSCs (monosynaptic IPSCAs) by inducing a positive shift in the reversal potential and a decrease in slope conductance. Responses to pressure-applied isoguvacine, a GABAA receptor agonist, were reversibly depressed by anoxia, again because of a positive shift in reversal potential and decrease in conductance. Anoxic effects on slope conductances and reversal potential of isoguvacine responses and monosynaptic IPSCA coincided, suggesting that evoked transmitter release from GABAergic terminals was not affected by anoxia. 8. Anoxic depression of monosynaptic GABAB receptor-mediated IPSCs (monosynaptic IPSCBs) was due to a decrease in the slope conductance of monosynaptic IPSCB. In contrast to EPSCs, DPCPX (200 nM) failed to prevent anoxia-induced depression of mIPSCA and mIPSCB. 9. Paired-pulse depression of monosynaptic IPSCs, partially mediated by presynaptic GABAB receptors, was not affected by anoxia. 10. These data provide direct evidence for the hypothesis that inhibitory interneurons of CA1 stratum LM are functionally disconnected from excitatory inputs by anoxia. This disconnection underlies the preferential block by anoxia of IPSCs recorded in pyramidal cells, and it may occult the postsynaptic modifications in GABAA and GABAB responses. This disconnection involves adenosine-dependent inhibition of glutamate release from excitatory terminals. GABA release and its modulation by presynaptic GABAB receptors, both known to be insensitive to adenosine, seems to be resistant to anoxia.

Animals↗
PubMed1995

Expression of LTP by AMPA and/or NMDA receptors is determined by the extent of NMDA receptors activation during the tetanus.

1. We have tested, in CA1 hippocampal slices, the hypothesis that the expression of long-term potentiation (LTP) by alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) and/or N-methyl-D-aspartate (NMDA) receptors depends on the degree of NMDA receptors activation during the tetanus. 2. Slices were perfused in an artificial cerebrospinal fluid (ACSF) containing glycine (1 microM), bicuculline (5 microM) and a low Mg2+ concentration (0.3 mM). To measure the AMPA and NMDA receptor-mediated field excitatory postsynaptic potential (fEPSPA and fEPSPN, respectively), we have used the following procedure: control fEPSPA was first measured, 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 10 microM) was then added, and fEPSPN was evoked. CNQX was washed, and once control fEPSPA was recorded, the Schaffer collaterals were tetanized at a weak or a strong intensity. The slope of fEPSPA was measured for 30-45 min followed by that of fEPSPN after the application of CNQX. 3. At a weak intensity (TW, which corresponds to a fEPSPA of approximately 0.3 mV of amplitude and no fEPSPN), the tetanic stimulation generated LTP of fEPSPA (58.7 +/- 8.1% mean +/- SE, n = 9) but no significant potentiation of the fEPSPN (11.2 +/- 2.2%, n = 9). These values were significantly different (P < 0.05, analysis of variance, Fisher test). 4. In 9 of 13 slices, tetanic stimulation of strong intensity (Ts, intensity corresponding to a fEPSPN of approximately 0.3 mV of amplitude) generated LTP of fEPSPN (89.1 +/- 17.2%) but not of fEPSPA (9.44 +/- 2.8%). In the four remaining slices the tetani induce LTP of both fEPSPA and fEPSPN (81.7 +/- 14.7% and 101 +/- 35.6%, respectively, both values were not significantly different). 5. We then examined the effects of decreasing fEPSPN by 50% in LTP generated by Ts and Tw. In the presence of 7-Chlorokynurenate (7Cl(-)-Kyn; 6 microM; n = 6), an antagonist of the allosteric glycine site of the NMDA receptors, Ts generated LTP of fEPSPA (63.2 +/- 8.2%) but not of fEPSPN (12.6 +/- 4.0%). Both values were significantly different. Tw still evoked LTP of fEPSPA but of smaller magnitude (29.8 +/- 6.3%, n = 8) than the one obtained in the absence of the antagonist (58.7 +/- 8.1%). Both values were significantly different. 6. The present observation suggests that l) LTP of fEPSPA has a lower threshold than that of fEPSPN, i.e., stronger activation of NMDA receptors during the tetani is required to induce LTP of fEPSPN than the one required for inducing LTP of fEPSPA; and 2) there is a bell-shaped relationship between the degree of activation of NMDA receptors during the tetani and the magnitude of LTP of the fEPSPA: tetani that generate LTP of fEPSPN have a low probability to induce LTP of fEPSPA. We suggest that AMPA and NMDA components are potentiated through two different presumably postsynaptic processes.

6-Cyano-7-nitroquinoxaline-2,3-dione↗
PubMed1994

Major differences between long-term potentiation and ACPD-induced slow onset potentiation in hippocampus.

We examined the effects of a long-lasting application of the selective metabotropic glutamate receptor (mGluR) agonist 1S-3R, 1-amino cyclopentane-1,3-dicarboxylic acid (ACPD) on synaptic potentials recorded from the CA1 and CA3 subfields in hippocampal slices maintained in a superfusion slice chamber. In 25% of the slices, ACPD generated an slow onset potentiation (SOP) of population EPSPs (pEPSPs) in CA1. In contrast to long-term potentiation (LTP) induced by a tetanic train, SOP was accompanied by an increase in the magnitude of the presynaptic fiber volley. Potentiation of the isolated afferent volley suggests that the expression of SOP is due to a recruitment of additional presynaptic fibers by the test stimulus caused by the persistent block of K+ channels by ACPD.

Action Potentials↗
PubMed1994

Benzodiazepines modulate calcium spikes in young and adult hippocampal cells.

GABAA receptor-independent effects of benzodiazepine receptor (BZ-R) agonists on calcium and barium spikes were recorded intracellularly in the presence of bicuculline from CA3 hippocampal cells of young and adult rats. Zolpidem (omega 1 BZ-R agonist), had no effects in young animals but decreased calcium and barium spikes and barium currents in adults. Midazolam (omega 1 and omega 2 BZ-R agonist) increased barium spikes in both young and adult animals. The effects on calcium spikes were more complex since a decrease was sometimes preceded by an increase. Thus, in adults zolpidem acting on omega 1 BZ-R reduced calcium influx while in young rats midazolam acting on omega 2 BZ-R increased barium influx. This modulation of calcium spikes by benzodiazepines could be relevant of calcium spikes by benzodiazepines could be relevant because of the developmental role played by calcium dependent processes.

Aging↗
PubMed1994

Proliferative astrocytes may express fibronectin-like protein in the hippocampus of epileptic rats.

Kainic acid treatment, a model of temporal lobe epilepsy, induces in CA3-CA4 fields of hippocampal complex a neuronal degeneration associated with glial hypertrophy and proliferation. After treatment with kainate, fibronectin (an extracellular matrix protein) immunoreactivity increases in CA3-CA4. Fibronectin antibodies stain proliferative cells (simultaneously labelled by [3H]thymidin) of astrocytic type (double-immunostained by GFAP antibodies). This result constitutes the first direct demonstration of astroglial fibronectin expression in vivo. In the molecular layer of kainate-treated rats there is an axon-terminal degeneration of association-fibers. This is associated with a transient hypertrophy of resident astrocytes but not with any glial proliferation. Reactive astrocytes do not express (or faintly) fibronectin immunoreactivity in this layer. Since fibronectin is involved in astroglial proliferation in vitro, the present observations suggest that astrocytes contribute in vivo to the astroglial proliferation by an autocrin mechanism.

Animals↗
PubMed1994

NMDA redox site modulates long-term potentiation of NMDA but not of AMPA receptors.

We have compared the effects of redox drugs on long-term potentiation mediated by AMPA or NMDA receptors. A reducing and an oxidizing agent had no effect on long-term potentiation mediated by AMPA receptors. In contrast, the induction of long-term potentiation mediated by NMDA receptors was prevented by a thiol oxidizing drug and restored by a disulfide reducing agent.

Animals↗
PubMed1994

Increased synthesis of specific proteins during glutamate-induced neuronal death in cerebellar culture.

We have previously shown that glutamate-induced neurotoxicity is mediated by a sodium-chloride component and a calcium component in our cerebellar granule cell culture. In order to further characterize these two different components, the time course of neuronal death induced by glutamate (100 microM) in basal solution and in low sodium-chloride solution was studied by morphological and biochemical criteria. As shown by phase-contrast microscopy, cerebellar granule cells exhibited clear neuronal degeneration within 4 h after exposure to this excitotoxin. These morphological changes correlated [35S]methionine incorporation into proteins which rapidly declined during the first hour of treatment. Qualitative change in [35S]methionine incorporation into proteins was further investigated by two-dimensional gel electrophoresis performed after glutamate exposure in basal solution and in low sodium-chloride solution. Most of the proteins showed a decreased labelling after glutamate exposure as expected, but some polypeptides showed an increased labelling or appeared to be newly synthesized. Furthermore, a different pattern of protein synthesis was observed when glutamate exposure was performed in basal solution or in low sodium-chloride solution. The identification of these polypeptides and their implication in the neuronal death are discussed.

Animals↗
PubMed1994

Development of mossy fiber synapses in hippocampal slice culture.

The mossy fiber synaptogenesis has been studied in hippocampal slice cultures. In vivo mossy fiber terminals contact the thorny excrescences of CA3 pyramidal neurons over a restricted portion, i.e. the proximal part of the apical dendrite. In organotypic cultures mossy fibers expand their terminal field and invade the infrapyramidal area of the CA3 region and the supragranular layer of the dentate gyrus. Newly formed mossy fiber synapses in CA3 region were examined, through electron microscopy, in cultures taken at various time intervals. The main events of the formation of newly formed mossy fiber synapses can be summarized as follows. During the first week following explantation mossy fiber axons contact the dendritic shaft of the pyramidal dendrite and establish both symmetrical and asymmetrical contacts. Subsequent modifications occur in the postsynaptic portion facing the mossy fiber bouton: (i) a massive accumulation of polyribosomes and coated vesicles in the subsynaptic cytoplasm; (ii) undulations of the plasma membrane; (iii) disappearance of neurotubules at postsynaptic sites and appearance of a fine network of filamentous material. Later on in culture, complex giant spines invaginate within the synaptic bouton. In conclusion this study shows that CA3 pyramidal neurons following deafferentation retain the capacity to form thorny excrescences, when contacted by mossy fibers. Moreover these results suggest a crucial role for mossy fibers to induce the formation of thorny excrescences in an heterotopic localization, i.e. over the basilar dendrites of CA3 pyramidal neurons.

Animals↗