Increase in slow afterhyperpolarization led to learning delay in DBA mice.

We showed that differences in learning capacity between DBA and C57Bl/6 mice correlates with differences in slow afterhyperpolarization amplitude in hippocampal CA1 pyramid neurons. In DBA mice learning capacity is lower, but the amplitude of slow afterhyperpolarizations higher than in C57Bl/6 mice.

Hippocampus-dependent learning and memory is impaired in mice lacking the Ras-guanine-nucleotide releasing factor 1 (Ras-GRF1).

Previous results have suggested that the Ras signaling pathway is involved in learning and memory. Ras is activated by nucleotide exchange factors, such as the calmodulin-activated guanine-nucleotide releasing factor 1 (Ras-GRF1). To test whether Ras-GRF1 is required for learning and memory, we inactivated the Ras-GRF1 gene in mice. These mutants performed normally in a rota-rod motor coordination task, and in two amygdala-dependent tasks (inhibitory avoidance and contextual conditioning). In contrast the mutants were impaired in three hippocampus-dependent learning tasks: contextual discrimination, the social transmission of food preferences, and the hidden-platform version of the Morris water maze. These studies indicate that Ras-GRF1 plays a role in hippocampal-dependent learning and memory.

Molecular, cellular, and neuroanatomical substrates of place learning.

Learning and remembering the location of food resources, predators, escape routes, and immediate kin is perhaps the most essential form of higher cognitive processing in mammals. Two of the most frequently studied forms of place learning are spatial learning and contextual conditioning. Spatial learning refers to an animal's capacity to learn the location of a reward, such as the escape platform in a water maze, while contextual conditioning taps into an animal's ability to associate specific places with aversive stimuli, such as an electric shock. Recently, transgenic and gene targeting techniques have been introduced to the study of place learning. In contrast with the abundant literature on the neuroanatomical substrates of place learning in rats, very little has been done in mice. Thus, in the first part of this article, we will review our studies on the involvement of the hippocampus in both spatial learning and contextual conditioning. Having demonstrated the importance of the hippocampus to place learning, we will then focus attention on the molecular and cellular substrates of place learning. We will show that just as in rats, mouse hippocampal pyramidal cells can show place specific firing. Then, we will review our evidence that hippocampal-dependent place learning involves a number of interacting physiological mechanisms with distinct functions. We will show that in addition to long-term potentiation, the hippocampus uses a number of other mechanisms, such as short-term-plasticity and changes in spiking, to process, store, and recall information. Much of the focus of this article is on genetic studies of learning and memory (L&M). However, there is no single experiment that can unambiguously connect any cellular or molecular mechanism with L&M. Instead, several different types of studies are required to determine whether any one mechanism is involved in L&M, including (i) the development of biologically based learning models that explain the involvement of a given mechanism in L&M, (ii) lesion experiments (genetics and pharmacology), (iii) direct observations during learning, and (iv) experiments where learning is triggered by turning on the candidate mechanism. We will show how genetic techniques will be key to unraveling the molecular and cellular basis of place learning.

Autophosphorylation at Thr286 of the alpha calcium-calmodulin kinase II in LTP and learning.

The calcium-calmodulin-dependent kinase II (CaMKII) is required for hippocampal long-term potentiation (LTP) and spatial learning. In addition to its calcium-calmodulin (CaM)-dependent activity, CaMKII can undergo autophosphorylation, resulting in CaM-independent activity. A point mutation was introduced into the alphaCaMKII gene that blocked the autophosphorylation of threonine at position 286 (Thr286) of this kinase without affecting its CaM-dependent activity. The mutant mice had no N-methyl-D-aspartate receptor-dependent LTP in the hippocampal CA1 area and showed no spatial learning in the Morris water maze. Thus, the autophosphorylation of alphaCaMKII at Thr286 appears to be required for LTP and learning.

Reduced K+ channel inactivation, spike broadening, and after-hyperpolarization in Kvbeta1.1-deficient mice with impaired learning.

A-type K+ channels are known to regulate neuronal firing, but their role in repetitive firing and learning in mammals is not well characterized. To determine the contribution of the auxiliary K+ channel subunit Kvbeta1.1 to A-type K+ currents and to study the physiological role of A-type K+ channels in repetitive firing and learning, we deleted the Kvbeta1.1 gene in mice. The loss of Kvbeta1.1 resulted in a reduced K+ current inactivation in hippocampal CA1 pyramidal neurons. Furthermore, in the mutant neurons, frequency-dependent spike broadening and the slow afterhyperpolarization (sAHP) were reduced. This suggests that Kvbeta1.1-dependent A-type K+ channels contribute to frequency-dependent spike broadening and may regulate the sAHP by controlling Ca2+ influx during action potentials. The Kvbeta1.1-deficient mice showed normal synaptic plasticity but were impaired in the learning of a water maze test and in the social transmission of food preference task, indicating that the Kvbeta1.1 subunit contributes to certain types of learning and memory.

Fast increases of AMPA receptor sensitivity following tetanus-induced potentiation in the CA1 region of the rat hippocampus.

Changes in sensitivity of the alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) subtype of glutamate receptors following the induction of long-term potentiation (LTP) were investigated in rat hippocampal slices. AMPA was ejected into the stratum radiatum of the hippocampal CA1 region by pressure ejection and the response recorded intracellularly. Responses to AMPA exhibited a fast increase (2-3 min) following LTP induction: the time course of this increase paralleled the potentiation of the excitatory postsynaptic current (EPSC). In cases where there was no increase in the slope of intracellularly recorded EPSC following tetanization, no increase of the AMPA responses was observed. These results suggest that increases in postsynaptic AMPA sensitivity can underlie short-term potentiation (STP) and thus, support a postsynaptic locus for both STP and LTP.

Antibodies to postsynaptic PKC substrate neurogranin prevent long-term potentiation in hippocampal CA1 neurons.

Both pre- and postsynaptic protein kinase C have been implicated in long-term potentiation. Neurogranin (also known as BICKs and RC3) is a neuronal postsynaptic protein kinase C substrate. In the present study we injected monoclonal IgGs that recognize the protein kinase C phosphorylation site in neurogranin and B-50 (GAP-43), and that have been shown to inhibit protein kinase C-mediated B-50 phosphorylation, through a whole-cell clamp pipette into CA1 pyramidal neurons in rat hippocampal slices. Injection of neurogranin IgGs, but not of control IgGs, prevented the induction of tetanus-induced long-term potentiation without affecting post-tetanic potentiation. Our results suggest that neurogranin is involved in mechanisms of activity-dependent synaptic plasticity.

Histaminergic modulation of synaptic plasticity in area CA1 of rat hippocampal slices.

The effects of histamine on baseline synaptic transmission and long-term potentiation (LTP) were investigated in the CA1 region of rat hippocampal slices. Bath applied histamine reversibly and dose-dependently increased the amplitude of extracellularly recorded population spikes in the concentration range 0.1-100 microM by a maximum of 40%. At higher concentrations (10-100 microM) histamine also caused a small depression of field excitatory postsynaptic potentials (fEPSPs) of approx 10%. The effect of histamine on population spikes was found to be mediated through histamine H2 receptors. Histamine (10-100 microM) was found to produce a statistically significant LTP of fEPSPs when combined with a weak tetanus (0.25 sec, 50 Hz). Histamine H1 (mepyramine, 1 microM) and H2 (cimetidine, 50 microM) receptor antagonists did not block this enhanced potentiation. In addition, histamine (10-100 microM) enhanced the late portion of the response produced by pressure ejection of glutamate receptor agonist N-methyl-D-aspartate into the slice, as recorded extracellularly or intracellularly. This effect of histamine was only apparent when large NMDA responses were obtained, using a high pipette concentration of NMDA (1 mM). In the presence of histamine H1 and H2 antagonists, potassium channel blockers or blockade of inhibition, this enhancement could still be observed. We conclude that histamine facilitated the induction of LTP, most likely by acting directly at the NMDA receptor.

Simultaneous local pressure microejection of excitatory amino acids and field potential recording with a single micropipette in the hippocampal slice.

The ability to use the same micropipette for simultaneous pressure microejection and field potential recording ('spot pressure electrode') is described. This approach allows the use of a single electrode to record both field responses to pressure microejection of drugs (fPR) and synaptically evoked field excitatory postsynaptic potentials (fEPSP). To demonstrate this, the glutamate receptor agonists alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA, 100-300 microM) and N-methyl-D-aspartate (NMDA, 1 mM) were used. Both AMPA- and NMDA-evoked fPRs exhibited properties of postsynaptic responses. Thus, they were not significantly altered by tetrodotoxin (TTX, 0.5-1 microM), but were completely and reversibly blocked by an antagonist of AMPA/kainate receptors, 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 20 microM) and D-2-amino-5-phosphonopentanoic acid (AP5, 20 microM), respectively. The responses to micropressure-ejected AMPA were more stable and probably restricted to a smaller locus than responses to iontophoretic AMPA ejection. Additionally, the AMPA and NMDA contained in the pipette (up to 500 microM) did not influence the fEPSP recorded with this pipette. Our results suggest that this method might be a useful experimental tool for studying the local effects of excitatory amino acids and other compounds.

An antagonist of glutamate metabotropic receptors, (RS)-alpha-methyl-4-carboxyphenylglycine, prevents the LTP-related increase in postsynaptic AMPA sensitivity in hippocampal slices.

(RS)-alpha-methyl-4-carboxyphenylglycine (MCPG), a compound which selectively antagonizes metabotropic glutamate receptors (mGlu-R), presents the LTP of field excitatory postsynaptic potentials (fEPSP) as well as the tetanus-induced increase in AMPA-evoked responses (fPRs) in the CA1 region of hippocampal slices. This effects of MCPG provide evidence for the involvement of mGlu-Rs in mechanisms underlying the postsynaptic maintenance of LTP, which appears to be mediated, at least partially, by an increase in sensitivity and/or number of postsynaptic AMPA receptors.

Priming stimulation facilitates Hebb-type plasticity in the Schaffer collateral-commissural pathways of the mouse hippocampus.

Hebb-type plastic changes of synaptic transmission were analyzed in the Schäffer collateral-commissural pathways (SC) of the mouse hippocampal slice. Low frequency (0.05-0.1 Hz) SC stimulation (S2) was paired with intracellular depolarization (100-200 ms; 2-3 nA) of CA1 pyramidal. Postpairing facilitation of synaptic responses in picrotoxin-free medium occurred only when S2 was "primed" with neurons the preceding shock (S1) administered through the same electrode 200 ms before S2. Priming-determined disinhibition in local circuitry is supposed to be responsible for this phenomenon.

Effects of philanthotoxin-343 on CA1 pyramidal neurons of rat hippocampus in vitro.

A synthetic analog of philanthotoxin-433, philanthotoxin-343 (PhTX-343), was tested in hippocampal pyramidal neurons in vitro. PhTX-343 (2 microM) did not significantly change synaptic transmission mediated by AMPA (alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid)/kainate receptors in the CA1 region of hippocampus. However, PhTX-343 significantly suppressed both the synaptic N-methyl-D-aspartate receptor-induced current (NMDA) obtained in the presence of CNQX(6-cyano-7-nitroquinoxaline-2,3-dione)/picrotoxin (10 microM) and the directly evoked NMDA receptor-induced current to pressure ejection of NMDA in the presence of tetrodotoxin (0.5 microM). A short transient facilitation of both types of NMDA response was seen immediately after the beginning of PhTX-343 application. Our results suggest that at high concentration (2 microM) PhTX-343 inhibits the NMDA-gated current, while the early facilitation occurred during an initial low concentration of the compound. Both facilitative and depressive actions of PhTX-343 are localized at the postsynaptic membrane.

[Effects of gamma-aminobutyric acid antagonist, 4-ethyl bicyclo-phosphate, on total and synaptic evoked responses in neurons of hippocampal slices]. / Vliianie antagonista gamma-aminomaslianoi kisloty 4-étilbitsiklofosfata na summarnye i sinapticheskie vyzvannye otvety neironov srezakh gippokampa.

Influence of 4-E-BPE on the amplitude of population spices (PS) evoked in CA1 area by Shaffer collateral stimulation in hippocampal slices were analysed. Bath application of 4-E-BPE (10(-6)-10(-5) M) led to a pronounced increase in the amplitude of the PS, the appearance of secondary PS and then introduction of GABA led to restoring original state. The 4-E-BPE was more potent than picrotoxin. These findings suggest that 4-E-BPE suppress inhibitory synaptic transmission in the CA1 region of hippocampus.

[The development of the system of recurrent inhibition in hippocampal field CAI in the early postnatal period]. / Razvitie sistemy vozvratnogo tormozheniia polia CAI gippokampa v rannem postnatal'nom periode.

Methods of focal potentials registration in the hippocampus sections have been used. CAI field during early postnatal ontogenesis has been studied.

[Tetrodotoxin--a sensitive component of the depolarizing response to GABA administration to the pyramidal neuron dendrites of the CA1 field of the hippocampus]. / Tetrodotoksin-chuvstvitel'nyi komponent depoliarizatsionnogo otveta na podvedenie GAMK k dendritam piramidnykh neironov polia CA1 gippokampa.

The intracellular recording of CA1 neurons in mouse hippocampal slice preparation was used to study the properties of depolarizing responses to iontophoretically applied GABA to their apical dendrites. Reversal potential of depolarizing responses was dependent on parameters of injecting current. It was about -60 mV and - (45-55) mV when iontophoretic currents 40-60 nA and 8-20 nA were used respectively. Application of tetrodotoxin (0.1-0.5 microM) resulted in decrease in amplitude of depolarizing responses evoked by weak currents, increase in slope of plot, reflecting relationship between response amplitude and membrane potential, and hyperpolarizing shift of reversal potential. Blocking++ of synaptic transmission with low calcium solution did not produce such changes. These results suggest that GABA depolarizing responses have a potential-sensitive component due to activation of sodium channels.

[Suppression of the "fast" IPSP when superimposed on the "slow" IPSP as a possible cause of priming in the mouse hippocampus]. / Podavlenie "bystrogo" TPSP pri nalozhenii ego na "medlennyi" TPSP kak vozmozhnaia prichina praiminga v gippokampe myshi.

Extra- and intracellular responses of the mouse hippocampus were recorded at CA1 region after stimulation of two independent inputs from the Schaffer collateral/commissural fibres: conditioning or priming input (C1) and testing or primed one (C2). Duration and amplitude of primed field potentials (FP) and excitatory postsynaptic potentials (EPSP) as well as amplitudes of early (IPSPa) and late (IPSPb) components of inhibitory postsynaptic potentials (IPSP) were measured with variation of C1-C2 intervals from 0 to 1 s. An increase in the FP duration as well as EPSP duration and amplitude and suppression of the IPSP amplitude occurred after conditioning with intervals of 50-500 ms, maximal effect was at 200 ms ("priming" effect). These changes correlated with the amplitude of priming IPSPb. The most prominent effect was observed in cells with hyperpolarizing IPSPa. It is assumed that primed FP and EPSP increase due to suppression of the primed IPSPa, when it is superimposed on the priming IPSPb.

[Effect of the gamma-aminobutyric acid antagonist disulfotetraazaadamantane on the evoked responses of neurons in hippocampal slices]. / Vliianie antagonista gamma-aminomaslianoi kisloty disul'fatetraazaadamantana na vyzvannye reaktsii neironov v srezakh gippokampa.

DSTA was analyzed for its effect, on the amplitude of population spikes (PS) evoked in CA1 area by the Shaffer collateral stimulation in hippocampal slices. Bath application of DSTA (10(-6)-10(-5) mol/l) caused a pronounced increase in the amplitude of PS and appearance of secondary PS. Introduction of GABA restored it to the initial state. These results demonstrate that DSTA inhibits the synaptic transmission in the CA1 region of the hippocampus.