High-speed two-photon imaging for in vivo analysis of brain disease
The objective of this proposal is to advance the method of two-photon fluorescence imaging for the analysis of animal models of brain disease. We intend to establish a two-tiered technical platform for high-speed imaging of brain activity correlates: We will optimize high-speed two-photon microscopy as well as develop new transgenic-viral labelling techniques to read out fast cellular events that underlie network dysfunction. Our particular focus will be on increasing the acquisition speed of two-photon imaging by a factor of 20 to 40 and improve labelling strength and specificity. This depends on coordinated efforts in the area of hardware improvement, software implementation and molecular reporter development. Optical investigations of altered signalling will be complemented by in vivo electrophysiology at the single cell and network levels and by behavioural analysis. We will use these techniques to investigate three important neurological disease states that are characterised by early alterations of fast neural activity: Alzheimer?s disease, where besides the expected silenced cells, hyperactive cortical neurons were recently discovered; epilepsy, where hyperactivation of neurons and glial cells is central, but the cellular and network mechanisms involved are poorly understood; and essential tremor, which serves as a model for the common neuronal hyper-synchronization observed in neurodegenerative movement disorders, such as Parkinson?s disease. Beside the obvious advantage of expanding the time resolution of two-photon imaging, the implementation of these technological advancements for the study of the diseased brain has the potential to foster understanding of the pathogenesis of a wide-variety of brain disorders characterized by network-dysfunction. Thus, the efforts of the 2p-Imaging consortium will provide technological advancement that will promote the development of new diagnostic and therapeutic tools in neurology.
Alzheimer's Disease, Imaging techniques, Molecular and genetic approaches, Gene targeting in the brain, Brain stimulation, Epilepsy, Essential tremor
2010 - 2013
Arthur Konnerth (Coordinator)
Marco de Curtis