Evaluation of Memantine in AAV-AD Rat: A Model of Late-Onset Alzheimer's Disease Predementia.

BACKGROUND: Though our understanding of Alzheimer's disease (AD) remains elusive, it is well known that the disease starts long before the first signs of dementia. This is supported by the large number of symptomatic drug failures in clinical trials and the increased trend to enroll patients at predementia stages with either mild or no cognitive symptoms. However, the design of pre-clinical studies does not follow this attitude, in particular regarding the choice of animal models, often irrelevant to mimic predementia Late Onset Alzheimer's Disease (LOAD). OBJECTIVES: We aimed to pharmacologically validate the AAV-AD rat model to evaluate preventive treatment of AD. METHODS: We evaluated an N-methyl-D-aspartate receptor antagonist, named memantine, in AAV-AD rats, an age-dependent amyloid rat model which closely mimics Alzheimer's pathology including asymptomatic and prodromal stages. Memantine was used at a clinically relevant dose (20 mg daily oral administration) from 4 (asymptomatic phase) to 10 (mild cognitive impairment phase) months of age. RESULTS: A 6-month treatment with memantine promoted a non-amyloidogenic cleavage of APP followed by a decrease in soluble Aß42. Consequently, both long-term potentiation and cognitive impairments were prevented. By contrast, the levels of hyperphosphorylated endogenous tau remained unchanged, indicating that a long-term memantine treatment is ineffective to restrain the APP processing-induced tauopathy. CONCLUSIONS: Together, our data confirm that relevant models to LOAD, such as the AAV-AD rat, can provide a framework for a better understanding of the disease and accurate assessment of preventive treatments.

Pre- and postsynaptic GABAB receptors in the hippocampus have different pharmacological properties.

Pharmacological properties of pre- and postsynaptic GABAB receptors were compared in CA1 hippocampal pyramidal neurons in vitro. The postsynaptic effects mediated by GABAB receptors, i.e., the baclofen-induced hyperpolarization, the bicuculline-resistant GABA response, and the slow inhibitory postsynaptic potential elicited by CA1 afferent stimulation, are all blocked by pertussis toxin (which inactivates some G proteins). These events are also suppressed by stimulating protein kinase C by phorbol esters and blocked by the selective GABAB antagonist phaclofen. In contrast, the baclofen-induced presynaptic depression of the excitatory postsynaptic potential elicited by CA1 afferent stimulation is resistant to the action of pertussis toxin and is not antagonized by phaclofen. However, this presynaptic inhibition can be antagonized by phorbol esters. These results indicate that the pre- and postsynaptic effects mediated by GABAB receptors in hippocampus have distinctly different pharmacological properties and possibly a different coupling mechanism.

A critical role for the glial-derived neuromodulator D-serine in the age-related deficits of cellular mechanisms of learning and memory.

Age-associated deficits in learning and memory are closely correlated with impairments of synaptic plasticity. Analysis of N-methyl-D-aspartate receptor (NMDAr)-dependent long-term potentiation (LTP) in CA1 hippocampal slices indicates that the glial-derived neuromodulator D-serine is required for the induction of synaptic plasticity. During aging, the content of D-serine and the expression of its synthesizing enzyme serine racemase are significantly decreased in the hippocampus. Impaired LTP and NMDAr-mediated synaptic potentials in old rats are rescued by exogenous D-serine. These results highlight the critical role of glial cells and presumably astrocytes, through the availability of D-serine, in the deficits of synaptic mechanisms of learning and memory that occur in the course of aging.

Age-related alterations of GABAergic input to CA1 pyramidal neurons and its control by nicotinic acetylcholine receptors in rat hippocampus.

The aim of this study was to determine whether age-associated alterations in the GABAergic input to pyramidal neurons in the hippocampus are due to a dysfunction of GABAergic interneurons, and/or a decrease in their cholinergic control via nicotinic receptors (nAChRs). Electrophysiological recordings were obtained from pyramidal cells in the CA1 area of hippocampal slices from young (3-4 months old) and aged (25-30 months old) Sprague-Dawley rats. Synaptic GABA(A) receptor-mediated inhibitory postsynaptic currents and inhibitory postsynaptic potentials induced by stimulation of the stratum oriens were significantly smaller in aged rats. The frequency (but not amplitude) of spontaneous and miniature GABA inhibitory postsynaptic currents (IPSCs) was reduced in aged rats, suggesting a presynaptic alteration. Tetanic stimulation of cholinergic afferents to release endogenous acetylcholine, or an exogenous application of the nAChR agonist cytisine, increased the frequency of spontaneous IPSCs in young rats; however these effects were not evident in aged rats, indicating that the nicotinic control of GABA release is lowered during aging. None of these age-related alterations were reversed by a chronic treatment with donepezil, a cholinesterase inhibitor. Immunofluorescent labeling of GABA interneurons with somatostatin (SOM), parvalbumin (PV) or calbindin (CB), together with the vesicular acetylcholine transporter VAChT, revealed a selective loss of subpopulations of SOM and CB positive interneurons. This loss was associated with a general decrease in density of the cholinergic network in aged rats. Thus, the lower GABAergic inhibition observed in the aged rat hippocampus is due to a selective loss/dysfunction of subpopulations of GABAergic interneurons, associated with a widespread cholinergic deficit.

Age-related alterations in the properties of hippocampal pyramidal neurons among rat strains.

We compared age-related alterations in the electrophysiological and pharmacological properties of CA1 hippocampal pyramidal neurons in three strains of rats (Sprague-Dawley, Fisher 344, and Wistar) at 3-4 and 25-32 months of age, using the in vitro slice preparation. The most consistent age-related alterations in the properties of rat hippocampal neurons were: a decrease in membrane excitability, a decrease in the amplitude and duration of inhibitory postsynaptic potentials and a decreased sensitivity to the effect of the cholinergic agonist carbachol. In contrast, no consistent alterations in calcium-dependent events were observed in these strains of rats. The age-related changes in the duration of the afterhyperpolarizations (AHPs) were different (and even opposite) depending on the strain studied. Our results show that age-related changes observed in a given strain are not necessarily present in all strains of the same species.

NMDA receptor activation in the aged rat: electrophysiological investigations in the CA1 area of the hippocampal slice ex vivo.

The effects of aging on activation of N-methyl-D-aspartate (NMDA) receptors were studied in the CA1 field of hippocampal slices from young (2-4 months old) and aged (25-32 months old) Sprague-Dawley rats with the use of ex vivo extra- and intracellular electrophysiological recording techniques. No significant age-related changes of the unitary NMDA-receptor mediated excitatory postsynaptic potentials (EPSPs), recorded from the pyramidal cells after stimulation of the stratum radiatum in a magnesium-free medium and isolated in the presence of the non-NMDA receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione, were found. Simultaneously, the magnitude of synaptic plasticity which involved NMDA receptor activation was not altered. No significant age-related modifications in the mechanisms controlling glutamate release and of postsynaptic NMDA receptor responsiveness were revealed. Considering the 30-40% decrease in NMDA binding sites in the aged hippocampus, our results suggest the occurrence of compensatory mechanisms which are discussed.

Loss of Calbindin-28K Immunoreactivity in Hippocampal Slices from Aged Rats: a Role for Calcium?

Calbindin-28K (CaBP) is a calcium-binding protein widely distributed in the brain. This protein appears to be involved in the sequestration and the translocation of intracellular free calcium. In this study, we have examined the distribution pattern of the structures immunoreactive for CaBP in the hippocampal formation from slices of young (4 months) and aged (24 - 27 months) rats previously submitted to electrophysiological measurements. We demonstrated a marked loss in the number of pyramidal cells immunoreactive for CaBP in aged rats as compared to young rats. A consistent decrease in the staining intensity was also revealed by optical density measurements. Some experiments have suggested that calcium homeostasis is modified in hippocampal neurons of aged rats. The loss of CaBP-like immunoreactivity (CaBP-LI) labelling could result from an increase in intracellular calcium concentrations. To support this hypothesis, we showed that in young rats (i) the CaBP-LI was enhanced in pyramidal neurons when the slice was preincubated in a calcium-free medium, and (ii) CaBP-LI was strongly decreased when the slice was preincubated in a high-calcium medium (5 mM) and when the entry of calcium into the cell was increased by a short application of an excitatory amino acid in the medium. Our results suggest that the loss of CaBP-LI in the hippocampus of aged rats could be due to an increase in intracellular calcium concentration. Preliminary observations of hippocampal slices at different times after induction of long-term potentiation (LTP) failed to show significant changes in CaBP immunoreactivity, suggesting that this calcium-binding protein is not directly involved in LTP processes.

Pharmacological characterization of an unusual mGluR-evoked neuronal hyperpolarization mediated by activation of GIRK channels.

The metabotropic glutamate receptor (mGluR) agonist ACPD exerts an unusual inhibitory effect on a population of neurons of the song-control nucleus HVc of the zebra finch via activation of the GIRK channel. We report in the present study the pharmacology of this response. ACPD directly hyperpolarized the neurons by a mechanism independent of GABA(B) receptors. The group I mGluR agonist DHPG had no effect on membrane properties and the group I mGluR antagonist 4-CPG did not affect the ACPD-induced hyperpolarization. In contrast, the ACPD response was mimicked by the group II mGluR agonist LY314593 and the group II and III agonist L-CCG-I. The group II mGluR antagonist LY307452 fully antagonized the ACPD response and reduced the response induced by L-CCG-I. The group III mGluR agonist L-AP4 induced a small hyperpolarization, which was antagonized by the group III mGluR antagonist MAP-4. These data indicate that group II and group III mGluRs are present and functional in the postsynaptic membrane of these HVc neurons, and mediate the hyperpolarizing action of mGluR agonists. In contrast, group I mGluRs are absent from these neurons, nonfunctional, or coupled to different effector systems that do not influence membrane potential or input resistance.

Decrease in calbindin content significantly alters LTP but not NMDA receptor and calcium channel properties.

The contribution of the cytosolic calcium binding protein calbindin D(28K) (CaBP) to the synaptic plasticity was investigated in hippocampal CA1 area of wild-type and antisense transgenic CaBP-deficient mice. We showed that long-term potentiation (LTP) induced by tetanic stimulation in CaBP-deficient mice was impaired. The fundamental biophysical properties of NMDA receptors and their number were not modified in CaBP-deficient mice. We also demonstrated that the physiological properties of calcium channels were identical between genotypes. An insufficient Ca(2+) entry through NMDA receptors or calcium channels, or a decrease in NMDA receptor density are unlikely to explain this impairment of LTP. Interestingly, we showed that the loss of LTP was not prevented by glycine but was restored in the presence of a low concentration of the NMDA receptor antagonist D-APV (5 microM) and of the calcium chelator BAPTA-AM (5 microM). Moreover, we observed a loss of LTP in the wild-type mice when the postsynaptic tetanic-induced [Ca(2+)](i) rise is excessively increased. Conversely, a weaker tetanus stimulation allowed LTP induction and maintenance in CaBP-deficient mice. These results suggest that a higher cytosol [Ca(2+)](i), due to the decrease of CaBP expression may impair LTP induction and maintenance mechanisms without affecting the mechanisms of calcium entry. Thus, CaBP plays a critical role in long term synaptic plasticity by limiting the elevation of calcium rise in the cytosol to some appropriate spatio-temporal pattern.

Effects of neuropeptides on rat cortical neurons: laminar distribution and interaction with the effect of acetylcholine.

The effects of the microiontophoretic application of five different peptides (cholecystokinin octapeptide sulfated form, cholecystokinin octapeptide non-sulfated form, vasoactive intestinal polypeptide, angiotensin-II and substance P) on cortical neurons were studied in rats anaesthetized with urethane. Vertical electrode penetrations were made in the first somatic sensory cortex and the laminar position of the neurons determined by the reconstruction of the tracks based on extracellular dye deposits. The first type of effect observed was an excitation of some cortical neurons. These neurons were mostly found in infragranular layers, specially in layer Vb. Pyramidal tract neurons were more often excited by peptides than the cortical population taken as a whole. Substance P excited the largest percentage of neurons, followed by vasoactive intestinal polypeptide and cholecystokinin octapeptide sulfated form, whereas angiotensin II and cholecystokinin octapeptide non-sulfated form were the least potent in terms of frequency of neurons excited as well as of amplitude of the responses. The vast majority of the neurons excited by a peptide could also be excited by acetylcholine. A second and independent effect of peptides was observed: the neuronal excitation induced by acetylcholine could be depressed by the simultaneous application of peptide. This depressing effect was also the most frequently observed with substance P, followed by cholecystokinin and vasoactive intestinal polypeptide.

Cortical projections of the nucleus of the diagonal band of Broca and of the substantia innominata in the rat: an anatomical study using the anterograde transport of a conjugate of wheat germ agglutinin and horseradish peroxidase.

The projections of the nucleus of the diagonal band of Broca and of the substantia innominata to the cerebral cortex were studied in the rat, using the anterograde transport of wheat germ agglutinin conjugated with horseradish peroxidase. Following diagonal band injections, fibers were observed ascending in the septum and reaching the cingulate cortex. They had a rostrocaudal, horizontal direction, mostly in layer VI and could be followed over long distances on sagittal sections. The fibers gave off collaterals which were seen ascending in the cerebral cortex and reaching more superficial layers. Following substantia innominata injections, fibers were observed to take two routes: the first one identical to the one described above and a second through the caudate-putamen reaching the temporo-insular cortex. Terminal fields had a more diffuse distribution following substantia innominata than following diagonal band injections. No clear laminar pattern of termination was observed. However the density of terminals was higher in layers IV, V and VI than in layers I, II and III. Since the conjugate used is not taken up by fibers of passage, this pattern of connection is believed to reflect the organization of the projection of the nucleus of the diagonal band and of the substantia innominata to the cerebral cortex.

Cerebral neocortical neurons in the aged rat: spontaneous activity, properties of pyramidal tract neurons and effect of acetylcholine and cholinergic drugs.

The properties of cortical cerebral neurons have been studied and compared in 2, 22 and 26 month-old Sprague-Dawley rats, using electrophysiological techniques. The mean spontaneous activity of the neurons in old animals (unidentified as well as pyramidal tract neurons) was not different from that of young adult rats. In contrast the mean latency of the antidromic response of pyramidal tract neurons to pyramidal tract stimulation was significantly longer in 26 month-old animals. No difference was observed in the effects of the excitatory amino acid glutamate applied by iontophoresis. The percentage of cortical neurons excited by the iontophoretic application of acetylcholine was similar in young and old animals. Neither the laminar distribution, nor the individual sensitivity of these neurons to acetylcholine were found to be modified. The pharmacological properties of the acetylcholine-induced excitations were unchanged, exhibiting muscarinic as well as nicotinic properties. These results are consistent with the suggestion that the impairment of the cholinergic system with aging is for a large part presynaptic. They also emphasize the fact that several physiological and pharmacological properties of the cerebral cortical neurons show little change with age in Sprague-Dawley rats.

Excitatory effect of acetylcholine on different types of neurons in the first somatosensory neocortex of the rat: laminar distribution and pharmacological characteristics.

In rats anaesthetized with either urethane, pentobarbital or fluothane the effects of acetylcholine, cholinergic agonists and antagonists (applied by iontophoresis) were studied on single cortical neurons of first somatosensory region. The laminar distribution of the neurons excited by acetylcholine was determined by the reconstruction of each electrode track based on a dye-deposit made at the last recording site. Neurons were identified using antidromic stimulation of the pyramidal tract, the ventrobasal thalamus and the corpus callosum. Neurons excited by acetylcholine could be segregated into two groups: one encompassing layer Vb and the upper part of layer VI, the other more deeply located at the limit between the cerebral cortex and the subjacent white matter. Neuronal responses to glutamate and nicotine, unlike those to actylcholine were evenly distributed in the cortex. Pyramidal tract neurons had corticothalamic neurons were frequently excited by acetylcholine and were shown to be located with the first group of acetylcholine sensitive neurons. Commissural neurons were rarely excited by acetylcholine and were not restricted to either group. The analysis of neuronal responses to acetylcholine and various agonists (carbachol, nicotine, acetyl-beta-methylcholine, carbamyl-beta-methylcholine, butyrylcholine) and antagonists (atropine, mecamylamine) revealed a prominent but not exclusive muscarine character. It is included (i) that cortical neurons of first somatosensory cortex which are excited by acetylcholine belong to two populations, one consisting, at least in part, of projection neurons (upper group) and the other of interneurons (lower group); (ii) that cortical acetylcholine receptors are of a 'mixed' type strongly weighted toward the muscarinic side.

Alterations in the properties of hippocampal pyramidal neurons in the aged rat.

The electrophysiological and pharmacological properties of CA1 hippocampal pyramidal neurons were studied in slices from young (three to four months) and aged (25-32 months) Sprague-Dawley rats having previously performed two behavioral tasks. About 20% of the aged rats were impaired in either the spontaneous alternation task or the water maze task. Electrophysiological parameters were measured and compared in young and aged animals using intracellular recordings. No age-related differences were observed in membrane potential, input resistance, amplitude of action potentials or amplitude of calcium spikes. The amplitude and duration of individual afterhyperpolarizations following a single spike were unchanged. In contrast, the neuronal excitability was significantly decreased and the spike duration significantly enhanced in aged rats as compared to young rats. The comparison of afterhyperpolarizations (which follow a burst of spikes) between young and aged rats was more complex. An increase in the amplitude and duration of afterhyperpolarizations generally occurred in aged animals. However, this increase was not consistent among animals and was dependent on the holding potential of the neuron and on the number of action potentials used to trigger the afterhyperpolarization. The depolarizing effect of bath-applied carbachol, as well as the associated increase in membrane resistance were reduced in neurons from aged rats. In contrast, the effects of carbachol on the depression of synaptic events and the blockade of the afterhyperpolarizations were similar in young and aged animals. In addition, the amplitude of the slow cholinergic excitatory postsynaptic potential induced by stimulation of cholinergic afferents in the presence of physostigmine was also decreased in aged rats. Excitatory postsynaptic potentials and inhibitory postsynaptic potentials following electrical stimulation of stratum radiatum were compared. The amplitude and duration of excitatory postsynaptic potentials were increased in aged rats. The amplitude and duration of the fast inhibitory postsynaptic potential were not significantly affected in aged animals. In contrast, the duration of the slow inhibitory postsynaptic potential was decreased in aged rats. Since the mean baclofen-induced hyperpolarization was only slightly reduced in aged rats, the most likely explanation is a decrease in the release of GABA rather than an alteration in the postsynaptic response mediated by GABAB receptors. A statistically significant correlation was found between the degree of impairment in the spontaneous alternation task and the amplitude of the carbachol-induced depolarization.

Effect of psychotropic drugs on identified septohippocampal neurons.

The effects of various psychotropic drugs (benzodiazepines, antidepressants, neuroleptics and nootropic drugs, a family of cognition activator agents) on firing rates of septohippocampal neurons, identified by electrical antidromic stimulation, were studied in the medial septum-nucleus of the diagonal band of Broca of rats anaesthetized with urethan. Extracellular potentials from single septohippocampal neurons were recorded using glass pipettes. Drugs were applied by either microiontophoresis or intravenous injections (i.v.). Benzodiazepines produced a marked depression of spontaneous firing rates of septohippocampal neurons whether applied i.v. (diazepam) or iontophoretically (flurazepam, midazolam). In addition, diazepam had a potent depressant effect on the rhythmically bursting activity of the septohippocampal neurons. Baclofen also had an inhibitory effect. Antidepressant drugs (applied by iontophoresis) as well as amphetamine, had a depressant effect on spontaneous firing rates. Neuroleptics (i.v.) had less significant or consistent effects on septohippocampal neurons, although the effects of haloperidol were usually inhibitory. Nootropic drugs were generally ineffective. These data indicate that most psychotropic drugs tested (with the exception of nootropic drugs) have an inhibitory effect on the spontaneous activity of septohippocampal neurons. However, benzodiazepines seem to be more active than antidepressants or neuroleptics. Oxotremorine (i.v.) had a potent excitatory effect on septohippocampal neurons. Atropine (i.v.) increased the septohippocampal neurons' firing rate in some cases. These results are discussed in view of the possible implication of the involvement of septohippocampal neurons in the mediation of the effects of psychotropic drugs on the central nervous system and, more specifically, on the cholinergic systems.

Spatial discrimination learning and CA1 hippocampal synaptic plasticity in mdx and mdx3cv mice lacking dystrophin gene products.

Duchenne muscular dystrophy is frequently associated with a non-progressive cognitive deficit attributed to the absence of 427,000 mol. wt brain dystrophin, or to altered expression of other C-terminal products of this protein, Dp71 and/or Dp140. To further explore the role of these membrane cytoskeleton-associated proteins in brain function, we studied spatial learning and ex vivo synaptic plasticity in the mdx mouse, which lacks 427,000 mol. wt dystrophin, and in the mdx3cv mutant, which shows a dramatically reduced expression of all the dystrophin gene products known so far. We show that reference and working memories are largely unimpaired in the two mutant mice performing a spatial discrimination task in a radial maze. However, mdx3cv mice showed enhanced emotional reactivity and developed different strategies in learning the task, as compared to control mice. We also showed that both mutants display apparently normal levels of long-term potentiation and paired-pulse facilitation in the CA1 field of the hippocampus. On the other hand, an increased post-tetanic potentiation was shown by mdx, but not mdx3cv mice, which might be linked to calcium-regulatory defects. Otherwise, immunoblot analyses suggested an increased expression of a 400,000 mol. wt protein in brain extracts from both mdx and mdx3cv mice, but not in those from control mice. This protein might correspond to the dystrophin-homologue utrophin. The present results suggest that altered expression of dystrophin or C-terminal dystrophin proteins in brain did not markedly affect hippocampus-dependent spatial learning and CA1 hippocampal long-term potentiation in mdx and mdx3cv mice. The role of these membrane cytoskeleton-associated proteins in normal brain function and pathology remains to be elucidated. Furthermore, the possibility that redundant mechanisms could partially compensate for dystrophins' deficiency in the mdx and mdx3cv models should be further considered.

Spatial learning and synaptic hippocampal plasticity in type 2 somatostatin receptor knock-out mice.

Somatostatin is implicated in a number of physiological functions in the CNS. These effects are elicited through the activation of at least five receptor subtypes. Among them, sst2 receptors appear the most widely expressed in the cortex and hippocampal region. However, the specific role of this somatostatin receptor subtype in these regions is largely undetermined. In this study, we investigated the role of the sst2 receptor in the hippocampus using mice invalidated for the sst2 gene (sst2 KO mice). Complementary experimental approaches were used. First, mice were tested in behavioral tests to explore the consequences of the gene deletion on learning and memory. Spatial discrimination learning in the radial maze was facilitated in sst2 KO mice, while operant learning of a bar-pressing task was slightly altered. Mice were then processed for electrophysiological study using the ex vivo hippocampal slice preparation. Extracellular recordings in the CA1 area showed an enhancement in glutamatergic (AMPA and NMDA) responses in sst2 KO mice which displayed an increase in the magnitude of the short-term potentiation and long-term depression. In contrast, long-term potentiation was not significantly altered. Taken together, these data demonstrate that somatostatin, acting via sst2 hippocampal receptors, may contribute to a global decrease in glutamate efficiency and consequently alter glutamate-dependent plasticity and spatial learning.

Involvement of a pertussis toxin-sensitive G-protein in the pharmacological properties of septo-hippocampal neurones.

1. The physiological and pharmacological properties of identified septo-hippocampal neurones (SHNs) have been studied in rats pretreated with the bacterial toxin, pertussis toxin (PTX). 2. In rats anaesthetized with urethane and pretreated with PTX, the axonal conduction velocity was unchanged while the mean spontaneous activity was significantly increased. 3. PTX pretreatment had no effect on responses of SHNs to the iontophoretic application of gamma-aminobutyric acid (GABA) and cholinoceptor agonists (acetylcholine or carbachol). 4. Baclofen and 5-hydroxytryptamine (5-HT), almost exclusively inhibitory in control rats, had little effect or an excitatory effect in PTX pretreated rats. 5. These results suggest the involvement of a pertussis toxin-sensitive G-protein in responses medicated by 5-HT and GABAB-receptors but not in responses mediated by cholinoceptors and GABAA-receptors in medial septum neurones projecting into the hippocampus.

NMDA receptor activation in the aged rat hippocampus.

Age-related alterations of N-methyl-D-aspartate receptor (NMDAr) activation were investigated in the CA1 field of hippocampal slices from young (3-6 months old) and aged (25-33 months old) Sprague-Dawley rats by using ex vivo extracellular electrophysiological recording techniques. NMDAr-mediated field excitatory postsynaptic potentials (fEPSPs) were induced by electrical stimulation of glutamatergic fibers in a magnesium (Mg(2+))-free medium supplemented with the non-NMDAr antagonist CNQX. The fEPSPs were significantly smaller in aged rats, whereas the response of presynaptic afferent fibers remained unaffected. No significant age-related differences were found in the ability of Mg(2+) to depress the magnitude of NMDAr-mediated fEPSPs. The responsiveness of postsynaptic NMDAr to the agonist was assessed in both groups of animals. No age-related differences were recorded either in the depolarizing effect of bath-applied NMDA or in the magnitude of the depolarization after altering extracellular Mg(2+) concentration. Finally, short-term potentiation (STP) of excitatory transmission was studied in young and aged rats considering the pivotal role of NMDAr in synaptic plasticity. No age-related alterations of the magnitude and the time course of STP in response to 10 or 30Hz conditioning stimulation were found. Because of the decrease in the magnitude of NMDAr-mediated synaptic transmission in aged animals, the absence of obvious modifications of synaptic plasticity suggests the occurrence of compensatory mechanisms that are discussed.

Pharmacological characterization of muscarinic responses in rat hippocampal pyramidal cells.

Intracellular recording from hippocampal CA1 pyramidal cells was used to characterize the pharmacological properties of muscarinic responses. Results obtained with the M1 antagonist pirenzepine and the M2 antagonist gallamine suggest that an M1 muscarinic receptor is involved in the muscarinic-induced membrane depolarization and blockade of the afterhyperpolarization (AHP). On the other hand, an M2 receptor may be involved in the cholinergic depression of the EPSP and the blockade of the potassium current termed the M-current. Pretreatment of hippocampi with pertussis toxin did not prevent any of the muscarinic responses suggesting that a pertussis toxin-sensitive G-protein is not involved. The M-current, in contrast to the other muscarinic actions, was unaffected by muscarinic agonists which are weak at increasing phosphoinositide (PI) turnover and actually blocked the action of full agonists. This finding suggests that stimulation of PI turnover may be involved in the blockade of the M-current. Although activation of protein kinase C with phorbol esters has little effect on the M-current, intracellular application of inositol trisphosphate did reduce the M-current. We were unable to establish any clear relationship between biochemical effector systems and the muscarinic receptor subtypes.