Mossy fiber-CA3 Pathway Research Articles

Hippocampal corticotropin-releasing hormone neurons support recognition memory and modulate hippocampal excitability.

Corticotropin-releasing hormone (CRH) signaling in the hippocampus has been established to be important for mediating the effects of stress on learning and memory. Given our laboratory’s recent characterization of a subset of hippocampal CRH neurons as a novel class of GABAergic interneurons, we hypothesized that these local GABAergic hippocampal CRH neurons may influence hippocampal function. Here we applied an array of molecular tools to selectively label and manipulate hippocampal CRH neurons in mice, in order to assess this interneuron population’s impact on hippocampus-dependent behaviors and hippocampal network excitability. Genetically-targeted ablation of hippocampal CRH neurons in vivo impaired object recognition memory and substantially enhanced the severity of kainic acid-induced seizures. Conversely, selective activation of CRH neurons in vitro suppressed the excitability of the mossy fiber-CA3 pathway. Additional experiments are needed to reconcile the functions of GABA and CRH signaling of hippocampal CRH neurons on hippocampal function. However, our results indicate that this interneuron population plays an important role in maintaining adaptive network excitability, and provide a specific circuit-level mechanism for this role.

CaMKII requirement for the persistence of in vivo hippocampal mossy fiber synaptic plasticity and structural reorganization

CaMKII has been proposed as a molecular substrate for long-term memory storage due to its capacity to maintain an active autophosporylated state even after the decay of the external stimuli. The hippocampal mossy fiber-CA3 pathway (MF-CA3) is considered as a relevant area for acquisition and storage of different learning tasks. MF-CA3 pathway exhibits a form of LTP characterized by a slow initial increase in the EPSP slope that is independent of NMDA receptors activation. Our previous studies show that application of high frequency stimulation sufficient to elicit MF-CA3 LTP produces structural reorganization, in a manner independent of LTP induction, at the stratum oriens of hippocampal CA3 area 7days after stimulation. However, the molecular mechanisms that underlie the maintenance of MF-CA3 LTP as well as the concomitant structural reorganization in this area remain to be elucidated. Here we show that acute microinfusion of myr-CaMKIINtide, a noncompetitive inhibitor of CaMKII, in the hippocampal CA3 area of adult rats during the late-phase of in vivo MF-CA3 LTP blocked its maintenance and prevented the accompanying morphological reorganization in CA3 area. These findings support the idea that CaMKII is a key molecular substrate for the long-term hippocampal synaptic plasticity maintenance.

Maintenance of long‐term potentiation in hippocampal mossy fiber—CA3 pathway requires fine‐tuned MMP‐9 proteolytic activity

Mechanisms of synaptic plasticity involve proteolytic activity mediated by a complex system of proteases, including members of metalloproteinase (MMP) family. In particular, MMP-9 is critical in LTP maintenance in the Schaffer collateral-CA1 pathway and in the acquisition of hippocampus-dependent memory. Recent studies from this laboratory revealed that in the mossy fiber-CA3 (MF-CA3) projection, where LTP induction and expression are largely presynaptic, MMPs blockade disrupts LTP maintenance and that LTP induction is associated with increased MMP-9 expression. Here we used acute brain slices from MMP-9 knock-out mice and transgenic rats overexpressing MMP-9 to determine how manipulations in endogenous MMP-9 affect LTP in the MF-CA3 projection. Both types of transgenic models showed a normal basal synaptic transmission and short-term plasticity. Interestingly, the maintenance of LTP induced in slices from knock-out mice and overexpressing rats was nearly abolished. However, in the presence of active MMP-9, a gradual fEPSP autopotentiation was observed and tetanization evoked a marked LTP in knock-out mice. Additionally, in MMP-9-treated slices from wild-type mice, fEPSP autopotentiation also occurred and partially occluded LTP. This indicates that exogenous protease can restore LTP in null mice whereas in the wild-type, MMP-9 excess impairs LTP. We expected that LTP maintenance in transgenic rats could be re-established by a partial MMP blockade but non-saturating concentrations of MMP inhibitor were ineffective. In conclusion, we demonstrate that LTP maintenance in MF-CA3 pathway requires fine-tuned MMP-9 activity and raises the possibility that altered MMP-9 level might be detrimental for cognitive processes as observed in some neuropathologies.

Corrigendum to “Long term potentiation affects intracellular metalloproteinases activity in the mossy fiber–CA3 pathway” [Molecular and Cellular Neuroscience 50/2 (2012) 147–159

Corrigendum to “Long term potentiation affects intracellular metalloproteinases activity in the mossy fiber–CA3 pathway” [Molecular and Cellular Neuroscience 50/2 (2012) 147–159

Myristoylated alanine rich C kinase substrate (MARCKS) heterozygous mutant mice exhibit deficits in hippocampal mossy fiber-CA3 long-term potentiation

The myristoylated alanine-rich C kinase substrate (MARCKS) is a primary protein kinase C (PKC) substrate in brain thought to transduce PKC signaling into alterations in the filamentous (F) actin cytoskeleton. Within the adult hippocampus, MARCKS is highly expressed in the dentate gyrus (DG)-CA3 mossy fiber pathway, but is expressed at low levels in the CA3-CA1 Schaffer collateral-CA1 pathway. We have previously demonstrated that 50% reductions in MARCKS expression in heterozygous Marcks mutant mice produce robust deficits in spatial reversal learning, but not contextual fear conditioning, suggesting that only specific aspects of hippocampal function are impaired by reduction in MARCKS expression. To further elucidate the role of MARCKS in hippocampal synaptic plasticity, in the present study we examined basal synaptic transmission, paired-pulse facilitation, post-tetanic potentiation, and long-term potentiation (LTP) in the hippocampal mossy fiber-CA3 and Schaffer collateral-CA1 pathways of heterozygous Marcks mutant and wild-type mice. We found that LTP is significantly impaired in the mossy fiber-CA3 pathway, but not in the Schaffer collateral-CA1 pathway, in heterozygous Marcks mutant mice, whereas basal synaptic transmission, paired-pulse facilitation, and post-tetanic potentiation are unaffected in both pathways. These findings indicate that a 50% reduction in MARCKS expression impairs processes required for long-term, but not short-term, synaptic plasticity in the mossy fiber-CA3 pathway. The implications of these findings for the role of the mossy fiber-CA3 pathway in hippocampus-dependent learning processes are discussed.

FK960, a novel potential anti-dementia drug, enhances high K +-evoked release of somatostatin from rat hippocampal slices

We have demonstrated that FK960 [ N-(4-acetyl-1-piperazinyl)- p-fluorobenzamide monohydrate], a novel putative anti-dementia drug of piperazine derivative, ameliorates memory deficits in a variety of animal models of dementia in rats and monkeys, and also augments long-term potentiation (LTP) in the mossy fiber-CA3 pathway in guinea-pig hippocampal slices. Our recent studies have further suggested that somatostatin activation could be a primary mechanism of the pharmacological action of FK960. To clarify the mode of action of FK960 on somatostatinergic neurotransmission, FK960 was examined for its effects on somatostatin release from rat hippocampal slices. FK960 significantly enhanced high K +-evoked release, but not basal release, of somatostatin with similar concentration-dependency to its LTP augmenting action. On the other hands, FK960 had no effects on the release of neurotransmitters such as acetylcholine, 5-HT, d-aspartate or GABA from hippocampal slices. Our results provide compelling evidence that FK960 exerts specific and facilitatory actions on neural mechanisms involved in the activity-dependent release of somatostatin from nerve terminals of the hippocampus. These results also strengthen the view that FK960 regulates cognitive functions and augments LTP through an activation of the somatostatinergic nervous system in the hippocampus.

FK960, a novel potential anti-dementia drug, augments long-term potentiation in mossy fiber-CA3 pathway of guinea-pig hippocampal slices

Our previous studies have demonstrated that FK960 (FR59960; N-(4-acetyl-1-piperazinyl)- p-fluorobenzamide monohydrate), a novel antidementia piperazine derivative, exerts beneficial effects on memory deficits in various animal models of amnesia in rats [M. Yamazaki, N. Matsuoka, N. Maeda, Y. Ohkubo, I. Yamaguchi, FK960 N-(4-acetyl-1-piperazinyl)- p-fluorobenzamide monohydrate ameliorates the memory deficits in rats through a novel mechanism of action, J. Pharmacol. Exp. Ther., 279 (1996) 1157–1173.] and in rhesus monkeys [N. Matsuoka, T.G. Aigner, FK960 [ N-(4-acetyl-1-piperazinyl)- p-fluorobenzamide monohydrate], a novel potential antidementia drug, improves visual recognition memory in rhesus monkeys: comparison with physostigmine, J. Pharmacol. Exp. Ther., 280 (1997) 1201–1209]. To clarify the synaptic mechanisms of its antiamnesic action, FK960 was investigated for its effects on the development of long-term potentiation (LTP) in guinea-pig hippocampal slices. The magnitude of LTP of population spike recorded in CA3 pyramidal neurons was significantly augmented by perfusing FK960 (10 −9–10 −6 M) for 25 min before and during tetanic stimulation of the mossy fibers, whereas the basal amplitude of population spikes before tetanus was hardly affected by the drug. The dose–response curve was bell-shaped with a maximal augmentation at 10 −7 M. Scopolamine (10 −6 M) per se had little effect on the magnitude of LTP in the mossy fiber-CA3 pathway, but significantly attenuated its enhancement by FK960 (10 −7 M). In hippocampal slices from animals treated with cysteamine (200 mg/kg, s.c.), which was shown to deplete the hippocampal somatostatin, FK960 (10 −7 M) hardly affected the LTP. These results suggest that FK960 enhances the magnitude of LTP in the mossy fiber-CA3 pathway through an activation of the cholinergic–somatostatinergic link in the hippocampal formation. Furthermore, it can be postulated that the drug regulates the cognitive function by modulating directly synaptic plasticity in the hippocampal neuronal network.

Interactions among paired-pulse facilitation and post-tetanic and long-term potentiation in the mossy fiber-CA3 pathway in rat hippocampus.

Long-term potentiation (LTP) observed at the synapses of mossy fiber-CA3 (MF-CA3) pathway differs from that observed at the Schaffer collateral-CA1 pathway (SC-CA1), in being independent of N-methyl-D-aspartate (NMDA) receptors. The induction and expression mechanisms of MF-CA3 LTP remain to be determined. We have compared the occurrence and magnitude of LTP with that of two other indicators of presynaptic plasticity, post-tetanic potentiation (PTP) and paired-pulse facilitation in control brain slices from young rats and in slices treated with phorbol-12, 13-diacetate (PDAc), a protein kinase C activator. Paired-pulse facilitation is a potentiation of the second of two responses at intervals of tens of milliseconds and is due to a presynaptic calcium increase. Tetanic stimulation of mossy fibers induced LTP is area CA3 in only 64% of slices. In those slices that showed LTP, the size of the PTP was significantly greater than in those slices that did not, and the degree of correlation between LTP and PTP amplitude overall was r = 0.7. The degree of paired-pulse facilitation before tetanic stimulation was also positively correlated to the occurrence and magnitude of LTP and PTP after tetanic stimulation. The correlation coefficient between PTP and PPF was 0.749 for all slices studied, while that between LTP and PPF was 0.835 overall. Application of PDAc potentiated synaptic transmission and abolished paired-pulse facilitation (control ratio of second to first response, 2.1; after PDAc ratio 0.8) and LTP. PTP was absent at the control stimulus intensity in PDAc, but was apparent if the stimulus intensity was reduced to give a response of the same amplitude as before administration of PDAc. Stable LTP was also accompanied by a marked decrease in paired-pulse facilitation. These data suggest that MF-CA3 LTP, PTP and paired-pulse facilitation share common mechanisms and are all at least primarily of presynaptic origin. The occurrence of large paired-pulse facilitation or PTP is a predictor of a preparation which will show LTP. It is likely that presynaptic [Ca2+]i is an essential factor in LTP, PTP and paired-pulse facilitation, as well as the potentiation induced by application of PDAc, but the factors which determine whether or not [Ca2+]i rises following these various stimuli are not clear from the techniques used in these investigations.

Protein kinase C activation is necessary but not sufficient for induction of long-term potentiation at the synapse of mossy fiber-CA3 in the rat hippocampus.

The involvement of protein kinase C in long-term potentiation was investigated in the mossy fiber-CA3 pathway in an in vitro slice preparation of rat hippocampus. Tetanic stimulation induced stable long-term potentiation in the mossy fiber-CA3 pathway which was not affected by N-methyl-D-aspartate receptor antagonists. Long-term potentiation was not induced in the presence of a protein kinase C inhibitor, sphingosine. Application of 1 microM phorbol-12, 13-diacetate, an activator of protein kinase C, potentiated the synaptic response by about 400% and this potentiation was completely reversible upon washing. Sphingosine blocked the potentiation when it was applied before protein kinase C activation by phorbol-12, 13-diacetate. However, sphingosine had no effect on the potentiation when it was applied after the synaptic response was potentiated to a plateau following phorbol-12,13-diacetate perfusion. Long-term potentiation and phorbol ester-induced potentiation were not additive when phorbol-12,13-diacetate was applied after induction of long-term potentiation, suggesting that long-term potentiation and phorbol-12, 13-diacetate activate the same protein kinase C pool. The enhanced response caused by phorbol-12,13-diacetate returned to the long-term potentiation level after wash-out of phorbol-12,13-diacetate. Thus the cellular changes underlying long-term potentiation are long-lasting or permanent, while those caused by phorbol-12,13-diacetate are not. However, if tetanic stimulation was induced during prolonged phorbol-12,13-diacetate application (1 h), a potentiation similar in amplitude to long-term potentiation was induced but the population response returned to the control pre-long-term potentiation level after 2 h of washing. The potentiation following tetanic stimulation during prolonged application of phorbol-12,13-diacetate was blocked in the presence of D-2-amino-5-phosphonovaleric acid, a N-methyl-D-aspartate receptor antagonist. Thus, in the presence of phorbol esters the N-methyl-D-aspartate-independent long-term potentiation is occluded but a transient potentiation appears, presumably due to hyperexcitability and activation of N-methyl-D-aspartate receptors in recurrent pathways of area CA3. Normal N-methyl-D-aspartate-independent long-term potentiation could be induced after the 2 h washout period and now was maintained. In conclusion, protein kinase C activation is essential but not sufficient for long-term potentiation in the mossy fiber-CA3 pathway and when stimulated by application of phorbol esters produces a large and reversible synaptic potentiation. These investigations show that long-term potentiation in CA3 is a complex event involving several steps, and that activation of protein kinase C is only one of them.

Roles of endogenous cholinergic neurons in the induction of long-term potentiation at hippocampal mossy fiber synapses

To evaluate the functional role of endogenous acetylcholine (ACh) in the induction of long-term potentiation (LTP) at mossy fiber-CA3 synapses, the influence of cholinergic hypofunction on it was investigated. Administration of a cholinergic neurotoxin, ethylcholine mustard aziridinium ion (AF64A; 5 nmol, i.c.v.), to guinea pigs one week prior to preparing slices resulted in a significant decrease in the magnitude of LTP, associated with a significant decrease in cholineacetyltransferase (ChAT) activity and the number of ChAT immunoreactive cells in the hippocampal slices. Bath-application of a cholinesterase inhibitor, physostigmine at 0.1 μM and 10 μM, attenuated and augmented, respectively, the magnitude of LTP in slices prepared from vehicle-treated animals (naive slices), whereas that in slices prepared from AF64A-treated animals (lesioned slices) was not significantly affected by physostigmine at any concentration tested. The induction of LTP in naive slices was inhibited or facilitated by a muscarinic M 1 antagonist pirenzepine (1 μM) and by an M 2 antagonist AF-DX 116 (1 μM) alone, respectively, whereas that in lesioned slices was not significantly changed by either of them. Furthermore, bath-applied carbachol (CCh) at 0.01–10 μM augmented the magnitude of LTP in lesioned slices, whereas the induction of LTP in naive slices was inhibited and facilitated by CCh at 0.01–0.1 μM and 1–10 μM, respectively, as reported previously. Such an augmentation of LTP by CCh was reversed by pirenzepine, but not by AF-DX 116. These observations suggest that AF64A induces the defect in ACh release and the hypofunction of M 2 receptors, but not of M 1, at least during the induction of LTP at mossy fiber-CA3 synapses. Thus, it is conceivable that endogenous ACh plays a facilitatory and inhibitory role through muscarinic M 1 and M 2 receptors in the induction of LTP in the mossy fiber-CA3 pathway.

Role of somatostatin in the augmentation of hippocampal long-term potentiation by FR121196, a putative cognitive enhancer.

N-(4-Acetyl-1-piperazinyl)-4-fluorobenzenesulfonamide (FR121196), a newly introduced putative cognitive enhancer of a derivative of piperazine, was investigated for its effects on long-term potentiation in guinea-pig hippocampal slices. The magnitude of long-term potentiation of population spikes recorded in CA3 pyramidal neurons was significantly augmented by perfusing FR121196 (10(-9)-10(-6) M) for 25 min before and during tetanic stimulation of the mossy fibers; the basal amplitude of population spikes before tetanus was hardly affected by the drug. The dose-response curve was bell-shaped with a maximal augmentation at 10(-7) M. Similar activity and bell-shaped dose-response curve were observed with methamphetamine (10(-8)-10(-6) M). Physostigmine (10(-8)-10(-6) M) also facilitated long-term potentiation of this pathway and the magnitude of augmentation was concentration-dependent. Scopolamine (10(-6) M) per se had little effect on the magnitude of long-term potentiation in the mossy fiber-CA3 pathway, but significantly attenuated its enhancement by FR121196 (10(-7) M) and physostigmine (10(-6) M), although it failed to influence that by methamphetamine (10(-7) M). In hippocampal slices from animals treated with cysteamine, which was shown to deplete hippocampal somatostatin, FR121196 (10(-7) M) hardly affected long-term potentiation generation, whereas physostigmine (10(-6) M) and methamphetamine (10(-7) M) augmented it significantly. These results suggest that FR121196 enhances the development of long-term potentiation in the mossy fiber-CA3 pathway through activation of somatostatinergic neurons in the hippocampal formation.

P-407 - Facilitatory action of FR121196, a novel cognitive enhancer, on long-term potentiation of the mossy fiber-CA3 pathway in guinea-pig hippocampal slices

P-407 - Facilitatory action of FR121196, a novel cognitive enhancer, on long-term potentiation of the mossy fiber-CA3 pathway in guinea-pig hippocampal slices

Effects of thyrotropin-releasing hormone and a related analog, CNK-602A, on long-term potentiation in the mossy fiber-CA3 pathway of guinea pig hippocampal slices

The effects of thyrotropin-releasing hormone (TRH) and a related analog, CNK-602A, that induces the release of catecholamines, on long-term potentiation (LTP) of the population spike in mossy fiber-CA3 pathways were investigated in guinea pig hippocampal slices. TRH augmented LTP of the population spike at concentrations of 10(-6)-10(-5) M. CNK-602A also augmented LTP at concentrations of 10(-6)-10(-5) M in a dose-dependent manner. LTP in slices of the hippocampus obtained from animals given 6-hydroxydopamine (6-OHDA) intraventricularly was significantly lower than that in non-treated animals. However, both TRH and CNK-602A (10(-6) M) augmented LTP in slices from 6-OHDA-treated animals. These findings suggest that TRH and CNK-602A augment LTP in the mossy fiber-CA3 pathway without activating noradrenergic and/or dopaminergic fibers.

Somatostatin augments long-term potentiation of the mossy fiber-CA3 system in guinea-pig hippocampal slices

The effect of exogenously applied somatostatin (1–14), which is one of the candidates of neuromodulators in the hippocampus, on long-term potentiation (LTP) was investigated in the CA1 and CA3 subfields of guinea-pig hippocampal slices. In the mossy fiber-CA3 pyramidal cell system, the magnitude of LTP of both population excitatory postsynaptic potential (pEPSP) and population spike was significantly augmented by somatostatin (10★−10★M) perfused before and during tetanic stimulation, which never affected basal amplitude of population spikes before tetanus. The enhancement of LTP by somatostatin lasted for at least one hour after washout. On the other hand, somatostatin at the most effective concentration (3.2 × 10★M) in the above described system failed to affect the magnitude of the LTP of population spikes in Schaffer collateral-CA1 pathway. The enhancing effect of somatostatin on LTP in the mossy fiber CA3 system was inhibited either by a muscarinic antagonist, scopolamine (10★ M), or a β-adrenoceptor antagonist, timolol (10★ M). These results suggest that somatostatin enhances the production of LTP in the mossy fiber-CA3 pathway of the guinea-pig hippocampus through the intervention of cholinergic and noradrenergic neurons.

O-238 - Somatostatin augments long-term potentiation of the mossy fiber-CA3 system in guinea-pig hippocampal slices.

The effect of exogenously applied somatostatin (1–14), which is one of the candidates of neuromodulators in the hippocampus, on long-term potentiation (LTP) was investigated in the CA1 and CA3 subfields of guinea-pig hippocampal slices. In the mossy fiber-CA3 pyramidal cell system, the magnitude of LTP of both population excitatory postsynaptic potential (pEPSP) and population spike was significantly augmented by somatostatin (10−7−10−6M) perfused before and during tetanic stimulation, which never affected basal amplitude of population spikes before tetanus. The enhancement of LTP by somatostatin lasted for at least one hour after washout. On the other hand, somatostatin at the most effective concentration (3.2 × 10−7M) in the above described system failed to affect the magnitude of the LTP of population spikes in Schaffer collateral-CA1 pathway. The enhancing effect of somatostatin on LTP in the mossy fiber CA3 system was inhibited either by a muscarinic antagonist, scopolamine (10−6 M), or a β-adrenoceptor antagonist, timolol (10−6 M). These results suggest that somatostatin enhances the production of LTP in the mossy fiber-CA3 pathway of the guinea-pig hippocampus through the intervention of cholinergic and noradrenergic neurons.

Electron microscopic localization of γ- and βII-subspecies of protein kinase C in rat hippocampus

The subcellular distributions of the γ- and βII-subspecies of protein kinase C (γ- and βII-PKC) were studied in the rat hippocampus by light and electron microscopic immunocytochemistry. Both subspecies were abundant in the hippocampus with distinct subcellular distributions. The immunoreactivity of γ-PKC was observed throughout the Ammon's horn, while intense βII-PKC immunoreactivity was observed predominantly in the CA1 region. γ-PKC was distributed diffusely through the cytoplasm of pyramidal cells from the perikarya to the dendritic spines. In contrast, βII-PKC was concentrated around the Golgi complex and present diffusely in distal dendrites, except for the dendritic spines. Neither PKC subspecies could be detected in the presynaptic terminal. The postsynaptic localization of γ- and βII-PKC in CA1 suggests that both PKC subspecies may correlate to long-term potentiation in the CA1 region contributing to the postsynaptic side. γ-PKC may have a specific function not only in CA1 but also in the mossy fiber-CA3 pathway at the postsynaptic side. βII-PKC may have another function concerning the Golgi complex in CA1.

Interleukin-1β inhibits long-term potentiation in the CA3 region of mouse hippocampal slices

The effect of recombinant human interleukin-1β (IL-1β) on long-term potentiation (LTP) in the mossy fiber-CA3 pathway of mouse hippocampal slice preparations was investigated. IL-1β significantly inhibited LTP in concentrations as low as 2.9 pM (50 pg/ml). This effect of IL-1β was blocked by concurrent application of 100 nM Lys-D-Pro-Thr, a tripeptide analogue of IL-1β. This is the first evidence that IL-1β can regulate LTP of synaptic transmission in the hippocampus.

O-079 - Effects of thyrotropin-releasing hormone and a related analog, CNK-602A, on long-term potentiation of the mossy fiber-CA3 pathway in guinea pig hippocampal slices

O-079 - Effects of thyrotropin-releasing hormone and a related analog, CNK-602A, on long-term potentiation of the mossy fiber-CA3 pathway in guinea pig hippocampal slices