Stéphane Pages

Sensory-evoked LTP driven by dendritic plateau potentials in vivo

Long-term synaptic potentiation (LTP) is thought to be a key process in cortical synaptic network plasticity and memory formation. Hebbian forms of LTP depend on strong postsynaptic depolarization, which in many models is generated by action potentials that propagate back from the soma into dendrites. However, local dendritic depolarization has been shown to mediate these forms of LTP as well. As pyramidal cells in supragranular layers of the somatosensory cortex spike infrequently, it is unclear which of the two mechanisms prevails for those cells in vivo. Using whole-cell recordings in the mouse somatosensory cortex in vivo, we demonstrate that rhythmic sensory whisker stimulation efficiently induces synaptic LTP in layer 2/3 (L2/3) pyramidal cells in the absence of somatic spikes. The induction of LTP depended on the occurrence of NMDAR (N-methyl-d-aspartate receptor)-mediated long-lasting depolarizations, which bear similarities to dendritic plateau potentials. In addition, we show that whisker stimuli recruit synaptic networks that originate from the posteromedial complex of the thalamus (POm). Photostimulation of channelrhodopsin-2 expressing POm neurons generated NMDAR-mediated plateau potentials, whereas the inhibition of POm activity during rhythmic whisker stimulation suppressed the generation of those potentials and prevented whisker-evoked LTP. Taken together, our data provide evidence for sensory-driven synaptic LTP in vivo, in the absence of somatic spiking. Instead, LTP is mediated by plateau potentials that are generated through the cooperative activity of lemniscal and paralemniscal synaptic circuitry.

Effect of the excitation pulse carrier frequency on the ultrafast charge recombination dynamics of donor-acceptor complexes: Stochastic simulations and experiments

The influence of the excitation pulse carrier frequency on the ultrafast charge recombination dynamics of excited donor-acceptor complexes has been explored both theoretically and experimentally. The theoretical description involves the explicit treatment of both the optical formation of the nuclear wave packet on the excited free energy surface and its ensuing dynamics. The wave packet motion and the electronic transition are described within the framework of the stochastic point-transition approach. It is shown that the variation of the pulse carrier frequency within the absorption band can significantly change the effective charge recombination dynamics. The mechanism of this phenomenon is analyzed and a semiquantitative interpretation is suggested. The role of the vibrational coherence in the recombination dynamics is discussed. An experimental investigation of the ultrafast charge recombination dynamics of two donor-acceptor complexes in valeronitrile also is presented. The decays of the excited state population were found to be highly nonexponential, the degree of non-exponentiality depending on the excitation frequency. For one complex, the charge recombination dynamics was found to slow down upon increasing the excitation frequency, while the opposite behavior was observed with the other complex. These experimental observations follow qualitatively the predictions of the simulations.

Dendrites In Vitro and In Vivo Contain Microtubules of Opposite Polarity and Axon Formation Correlates with Uniform Plus-End-Out Microtubule Orientation

In cultured vertebrate neurons, axons have a uniform arrangement of microtubules with plus-ends distal to the cell body (plus-end-out), whereas dendrites contain mixed polarity orientations with both plus-end-out and minus-end-out oriented microtubules. Rather than non-uniform microtubules, uniparallel minus-end-out microtubules are the signature of dendrites in Drosophila and Caenorhabditis elegans neurons. To determine whether mixed microtubule organization is a conserved feature of vertebrate dendrites, we used live-cell imaging to systematically analyze microtubule plus-end orientations in primary cultures of rat hippocampal and cortical neurons, dentate granule cells in mouse organotypic slices, and layer 2/3 pyramidal neurons in the somatosensory cortex of living mice. In vitro and in vivo, all microtubules had a plus-end-out orientation in axons, whereas microtubules in dendrites had mixed orientations. When dendritic microtubules were severed by laser-based microsurgery, we detected equal numbers of plus- and minus-end-out microtubule orientations throughout the dendritic processes. In dendrites, the minus-end-out microtubules were generally more stable and comparable with plus-end-out microtubules in axons. Interestingly, at early stages of neuronal development in nonpolarized cells, newly formed neurites already contained microtubules of opposite polarity, suggesting that the establishment of uniform plus-end-out microtubules occurs during axon formation. We propose a model in which the selective formation of uniform plus-end-out microtubules in the axon is a critical process underlying neuronal polarization. SIGNIFICANCE STATEMENT Live-cell imaging was used to systematically analyze microtubule organization in primary cultures of rat hippocampal neurons, dentate granule cells in mouse organotypic slices, and layer 2/3 pyramidal neuron in somatosensory cortex of living mice. In vitro and in vivo, all microtubules have a plus-end-out orientation in axons, whereas microtubules in dendrites have mixed orientations. Interestingly, newly formed neurites of nonpolarized neurons already contain mixed microtubules, and the specific organization of uniform plus-end-out microtubules only occurs during axon formation. Based on these findings, the authors propose a model in which the selective formation of uniform plus-end-out microtubules in the axon is a critical process underlying neuronal polarization.

Single cell electroporation for longitudinal imaging of synaptic structure and function in the adult mouse neocortex in vivo

[This corrects the article on p. 36 in vol. 9, PMID: 25904849.].

Femtosecond time-resolved studies on bimolecular electron transfer processes

Bimolecular photo-induced electron transfer between an electron donor and an electron acceptor in a polar solvent may result in the formation of free ions (FI). Weller and coworkers have invoked several types of intermediates for describing this process exciplex or contact ion pair (CIP), loose ion pair (LIP), also called solvent separated ion pair. The knowledge of the structures of these intermediates is fundamental for understanding the details of bimolecular reactions in solution. However, up to now, no spectroscopic technique has been able to differentiate them. The UV-Vis absorption spectra of the ion pairs and the free ions are very similar. Furthermore, previous time resolved resonant Raman investigations have shown that these species exhibit essentially the same high frequency vibrational spectrum. Therefore, a pump/pump-probe experiment is developed to obtain more informations on the structures of these geminate ion pairs. It allows the investigation of the excited states dynamics of the transient species at different time delays after photo triggering the charge transfer, by monitoring the ground state recovery (GSR) of those transient species. This chapter uses perylene (Pe) as fluorescer (electron donor) and either trans-1,2-dicyanoethylene (DCE) or 1,4-dicyanobenzene (DCB) as quencher (electron acceptor) in acetonitrile (ACN).