Authors

Hay, Y. Audrey Andjelic, Sofija Badr, Sammy Lambolez, Bertrand

Affiliations

Neuroscience Paris Seine, Universite Pierre et Marie Centre National de la Recherche Scientifique
(CNRS), Paris, France, Institute National de la Sante et de la Recherche Me ´dicale (INSERM), Paris, France.

Application Area

Optogenetics

Electrophysiological Techniques

Abstract

Neocortical layer VI is critically involved in thalamocortical activity changes during the sleep/wake cycle. It receives dense projections from thalamic nuclei sensitive to the wake-promoting neuropeptides orexins, and its deepest part, layer VIb, is the only cortical lamina reactive to orexins. This convergence of wake-promoting inputs prompted us to investigate how layer VIb can modulate cortical arousal, using patch-clamp recordings and optogenetics in rat brain slices. We found that the majority of layer VIb neurons were excited by nicotinic agonists and orexin through the activation of nicotinic receptors containing α4-α5-β2 subunits and OX2 receptor, respectively. Specific effects of orexin on layer VIb neurons were potentiated by low nicotine concentrations and we used this paradigm to explore their intracortical projections. Co-application of nicotine and orexin increased the frequency of excitatory post-synaptic currents in the ipsilateral cortex, with maximal effect in infragranular layers and minimal effect in layer IV, as well as in the contralateral cortex. The ability of layer VIb to relay thalamocortical inputs was tested using photostimulation of channelrhodopsin-expressing fibres from the orexin-sensitive rhomboid nucleus in the parietal cortex. Photostimulation induced robust excitatory currents in layer VIa neurons that were not pre-synaptically modulated by orexin, but exhibited a delayed, orexin-dependent, component. Activation of layer VIb by orexin enhanced the reliability and spike-timing precision of layer VIa responses to rhomboid inputs. These results indicate that layer VIb acts as an orexin-gated excitatory feed forward loop that potentiates thalamocortical arousal.

Extract

… “ChR2-YFP-expressing neuronal somata and fibres were visualized with a 535-nm LED (CoolLed, Andover, UK)”…

Product Associated Features

pE-100: A range of compact, simple-to-use, single wavelength illumination systems for screening fluorescence.

Diascopic Technique

-

Live Cell Issues

-

Product Type

pE-100

Journal

Brain Structure and Function

Year of Publication

2014

Country of Publication

Germany