Phosphatase and tensin homolog, deleted on chromosome 10 deficiency in brain causes defects in synaptic structure, transmission and plasticity, and myelination abnormalities.

The phosphatidylinositol 3-kinase (PI3K) signaling pathway modulates growth, proliferation and cell survival in diverse tissue types and plays specialized roles in the nervous system including influences on neuronal polarity, dendritic branching and synaptic plasticity. The tumor-suppressor phosphatase with tensin homology (PTEN) is the central negative regulator of the PI3K pathway. Germline PTEN mutations result in cancer predisposition, macrocephaly and benign hamartomas in many tissues, including Lhermitte-Duclos disease, a cerebellar growth disorder. Neurological abnormalities including autism, seizures and ataxia have been observed in association with inherited PTEN mutation with variable penetrance. It remains unclear how loss of PTEN activity contributes to neurological dysfunction. To explore the effects of Pten deficiency on neuronal structure and function, we analyzed several ultra-structural features of Pten-deficient neurons in Pten conditional knockout mice. Using Golgi stain to visualize full neuronal morphology, we observed that increased size of nuclei and somata in Pten-deficient neurons was accompanied by enlarged caliber of neuronal projections and increased dendritic spine density. Electron microscopic evaluation revealed enlarged abnormal synaptic structures in the cerebral cortex and cerebellum. Severe myelination defects included thickening and unraveling of the myelin sheath surrounding hypertrophic axons in the corpus callosum. Defects in myelination of axons of normal caliber were observed in the cerebellum, suggesting intrinsic abnormalities in Pten-deficient oligodendrocytes. We did not observe these abnormalities in wild-type or conditional Pten heterozygous mice. Moreover, conditional deletion of Pten drastically weakened synaptic transmission and synaptic plasticity at excitatory synapses between CA3 and CA1 pyramidal neurons in the hippocampus. These data suggest that Pten is involved in mechanisms that control development of neuronal and synaptic structures and subsequently synaptic function.

Postsynaptic depolarisation enhances transmitter release and causes the appearance of responses at "silent" synapses in rat hippocampus.

Recent data indicate that most "silent" synapses in the hippocampus are "presynaptically silent" due to low transmitter release rather than "postsynaptically silent" due to "latent" receptors of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid type (AMPARs). That synapses bearing only N-methyl-d-aspartate (NMDAR) receptors do exist is suggested by the decreased number of transmission failures during postsynaptic depolarisation and by the presence of NMDA-mediated excitatory postsynaptic currents (EPSCs) in synapses silent at rest. We tested whether these effects could be due to potentiated transmitter release at depolarised postsynaptic potentials rather than removal of Mg(2+) block from NMDARs. Using whole-cell recordings of minimal EPSCs from CA1 and CA3 neurones of hippocampal slices we confirmed decreased incidence of failures at +40 mV as compared with -60 mV. This effect was associated with a gradual increase of EPSC amplitude after switching to +40 mV and with a decrease of paired-pulse facilitation. In initially silent synapses, potentiation of pharmacologically isolated AMPAR-mediated EPSCs was still observed at +40 mV and this persisted after stepping back to -60 mV. All above effects were blocked when the cell was dialysed with the Ca(2+) chelator BAPTA (20 mM). These observations are difficult to reconcile with the "latent AMPAR" hypothesis and suggest an alternative explanation, namely that the reduction in failure rates at positive potentials is due to potentiation of transmitter release following Ca(2+) influx through NMDARs. Our results suggest that silent synapses can be mainly "presynaptically" rather than "postsynaptically silent" and thus increased transmitter release rather than insertion of AMPARs is a major mechanism of early long-term potentiation maintenance.

Low-frequency depression of synaptic responses recorded from rat visual cortex.

To characterize the low-frequency depression (LFD) of synaptic transmission in the visual cortex, we recorded field potentials and minimal excitatory postsynaptic potentials (EPSPs) from layer II/III following intracortical stimulation at various frequencies in cortical slices of rats. Field potentials were stable at 0.017 Hz, but showed an amplitude depression at 0.033-0.1 Hz at stimulus intensity of 1.5 times the threshold for induction of the postsynaptic component and at 0.1-0.2 Hz at intensity of 1.2 times the threshold. The LFD was input-specific and its magnitude correlated with the stimulus frequency. An interruption of stimulation for 15 min yielded a nearly complete recovery from LFD. Minimal EPSPs tested at 0.1-1.7 Hz often showed LFD with similar features. However, some inputs were stable or even facilitated during repeated stimulation. At 0.1 and 0.2 Hz, >50% of inputs were stable, whereas 10% and 25% were depressed, respectively. At 0.5 and 1.7 Hz, LFD was observed in >60% and 80% of inputs, respectively. The magnitude of LFD strongly varied across inputs. In 3 of the 41 inputs analyzed, LFD was so strong that these inputs became virtually silent. Occurrence of responses to the second pulse in the paired-pulse paradigm when the first response was absent and recovery of depressed EPSPs following stimulus interruption or shift to a lower frequency suggest that these synapses were presynaptically silent due to a lowered probability of transmitter release. Altogether, the results indicate that testing intervals of <10 or even < or =30 s cannot be regarded as completely neutral. At the single-cell level, frequency-dependent changes were strongly heterogeneous across different inputs. LFD and its spontaneous recovery may underlie the previously described "post-rest" potentiation, and should be taken into account when considering information processing in cortical networks.

Long-term potentiation of the AMPA and NMDA components of minimal postsynaptic currents in rat hippocampal field Ca1.

The aim of the present work was to study the potentiation of the AMPA and NMDA components of minimal excitatory postsynaptic currents (EPSC) evoked by activation of restricted numbers of synapses. EPSC of neurons in field CA1 in hippocampal slices were recorded in whole-call patch-clamp conditions selected such that both (AMPA and NMDA) components were present, and these were measured in parallel using computational methods in combination with pharmacological receptor blockade. There was a quite strong correlation between the amplitudes of the AMPA and NMDA components and this was regarded as evidence that they were generated by the same synapses. In cases producing this correlation, both components showed essentially equal long-term potentiation lasting from 5 min to 2 h after afferent tetanization. The data did not support the postsynaptic hypothesis and were in better agreement with the concept that the major mechanism for the persistence of the initial phase of long-term potentiation (up to 1-2 h) is based on increases in the quantity of transmitter released presynaptically.

Selective suppression of forward and recurrent "rapid" inhibition by local application of picrotoxin in area CA1 of rat hippocampal slices.

Living slices of rat hippocampus were used to study the possibility of suppressing forward and recurrent "rapid" inhibition in area CA1 by local application of picrotoxin, an antagonist of ionotropic gamma-aminobutyric acid (GABAA) receptors. Application of picrotoxin to the apical dendrites increased the duration of focal potentials recorded in the radial layer (143.0 +/- 7.5%, n = 5; here and subsequently, results are presented as mean +/- error of the mean and n is the number of experiments) but had no effect on the population peak in the pyramidal layer (103.0 +/- 19.6%, n = 5). This is evidence for the existence of suppression of direct, but not of recurrent inhibition. Application of picrotoxin to the cell body layer, on the other hand, significantly increased the population peak (654.5 +/- 245.1%, n = 4) and provoked convulsive activity in neurons, demonstrating suppression of recurrent inhibition. Local application of picrotoxin was further used to study the question of how completely antagonists of glutamate ionotropic receptors sensitive to alpha-amino-3-hydroxy-S-methyl-4-isoxazole propionic acid (AMPA) suppress inhibition in solutions with low magnesium contents. This question is important for interpreting experimental data obtained from measurements of the components of excitatory postsynaptic potentials (EPSP), which depend on activation of ionotropic glutamate receptors sensitive to N-methyl-d-aspartic acid (NMDA). A number of studies have suggested that even at low concentrations, AMPA receptor antagonists suppress forward inhibition to such an extent that it has no significant effect on measurements of the NMDA component of EPSP. Our data do not contradict this suggestion.