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In vivo two-photon voltage-sensitive dye imaging reveals top-down control of cortical layers 1 and 2 during wakefulness.

ABSTRACT: Conventional methods of imaging membrane potential changes have limited spatial resolution, particularly along the axis perpendicular to the cortical surface. The laminar organization of the cortex suggests, however, that the distribution of activity in depth is not uniform. We developed a technique to resolve network activity of different cortical layers in vivo using two-photon microscopy of the voltage-sensitive dye (VSD) ANNINE-6. We imaged spontaneous voltage changes in the barrel field of the somatosensory cortex of head-restrained mice and analyzed their spatiotemporal correlations during anesthesia and wakefulness. EEG recordings always correlated more strongly with VSD signals in layer (L) 2 than in L1. Nearby (<200 mum) cortical areas were correlated with one another during anesthesia. Waking the mouse strongly desynchronized neighboring cortical areas in L1 in the 4- to 10-Hz frequency band. Wakefulness also slightly increased synchrony of neighboring territories in L2 in the 0.5- to 4.0-Hz range. Our observations are consistent with the idea that, in the awake animal, long-range inputs to L1 of the sensory cortex from various cortical and thalamic areas exert top-down control on sensory processing.

SUBMITTER: Kuhn B

PROVIDER: S-EPMC2396672 | biostudies-literature | 2008 May

REPOSITORIES: biostudies-literature

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In vivo two-photon voltage-sensitive dye imaging reveals top-down control of cortical layers 1 and 2 during wakefulness.

Kuhn B B Denk W W Bruno R M RM

Proceedings of the National Academy of Sciences of the United States of America 20080501 21

Conventional methods of imaging membrane potential changes have limited spatial resolution, particularly along the axis perpendicular to the cortical surface. The laminar organization of the cortex suggests, however, that the distribution of activity in depth is not uniform. We developed a technique to resolve network activity of different cortical layers in vivo using two-photon microscopy of the voltage-sensitive dye (VSD) ANNINE-6. We imaged spontaneous voltage changes in the barrel field of ...[more]

PMID: 18508976

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Project description:Optical recording techniques were applied to the turtle cerebellum to localize synchronous responses to microstimulation of its cortical layers and reveal the cerebellum's three-dimensional processing. The in vitro yet intact cerebellum was first immersed in voltage-sensitive dye and its responses while intact were compared to those measured in thick cerebellar slices. Each slice is stained throughout its depth, even though the pial half appeared darker during epi-illumination and lighter during trans-illumination. Optical responses were shown to be mediated by the voltage-sensitive dye because the evoked signals had opposite polarity for 540- and 710-nm light, but no response to 850-nm light. Molecular layer stimulation of the intact cerebellum evoked slow transverse beams. Similar beams were observed in the molecular layer of thick transverse slices but not sagittal slices. With low currents, beams in transverse slices were restricted to sublayers within the molecular layer, conducting slowly away from the stimulus site. These excitatory beams were observed nearly all the way across the turtle cerebellum, distances of 4-6mm. Microstimulation of the granule cell layer of both transverse or sagittal slices evoked a local membrane depolarization restricted to a radial wedge, but these radial responses did not activate measurable molecular layer beams in transverse slices. White matter microstimulation in sagittal slices (near the ventricular surface of the turtle cerebellum) activated the granule cell and Purkinje cell layers, but not the molecular layer. These responses were nearly synchronous, were primarily caudal to the stimulation, and were blocked by cobalt ions. Therefore, synaptic responses in all cerebellar layers contribute to optical signals recorded in intact cerebellum in vitro (Brown and Ariel, 2009). Rapid radial signaling connects a sagittally-oriented, fast-conduction system of the deep layers with the transverse-oriented, slow-conducting molecular layer, thereby permitting complex temporal processing between two tangential but orthogonal paths in the cerebellar cortex.

Dataset Information

In vivo two-photon voltage-sensitive dye imaging reveals top-down control of cortical layers 1 and 2 during wakefulness.

Publications

In vivo two-photon voltage-sensitive dye imaging reveals top-down control of cortical layers 1 and 2 during wakefulness.

Similar Datasets

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