Synapse to nucleus communication in intact cortical circuits
In order to faithfully respond and adapt to changes in the environment, synaptic activity must be integrated and communicated to the nucleus. Previous work has identified that strong synaptic activity in vitro can induce translocation of numerous signaling proteins from synapses to the nucleus. However, the extent and mechanisms of in vivo translocation and signaling to the nucleus following physiological patterns of synaptic activity are scarce. In this project, we will examine if and how specific synaptic/nuclear proteins undergo nuclear translocation and their role in experience-dependent sensory stimuli. To explore this, we will utilize newly developed gene editing tools, which allow genomic labeling using CRISPR/Cas9 in single neurons in the brain. This technique will enables to directly monitor innate intracellular signaling dynamics which underlie communication between synapse to nucleus during experience dependent plasticity.
Synapse
Nucleus
In utero electroporation of dividing cortical cells allows CRISPR Cas9 endogenous protein visualization in the brain. Images of HA-tagged CaMKIIg at synapses and CREB at the nucleus in cortical neurons.
Dynamics of epigenetics regulation in the brain
The neuronal genome undergoes widespread epigenetic regulation which is critical for normal brain development and adult brain plasticity. The importance of this modulation is demonstrated by many devastating neurological disorders and cognitive deficiencies which have been genetically associated with epigenetic factors and transcription modulators. We will utilize and develop new experimental approaches to allow dynamic visualization of epigenetic signaling in the brain. This will allow us to directly link ongoing epigenetic, neuronal activity and behavioral states, and to understand their physiological role. Ultimately, deciphering epigenetic signaling rules at the healthy brain will enable shed light on how its dysfunction leads to devastating cognitive decline and neurological disorders.
MeCP2-GFP expression in L2/3 neurons
Development of new techniques for monitoring and manipulation of neuronal signaling in living mice
We are working to develop new approaches to dissect neuronal signaling:
-New 2pFLIM biosensors for important signaling proteins.
-Methods for tagging of endogenous proteins in the brain using CRISPR/Cas9..
-Manipulation of protein function and expression via optogenetic control of important signaling pathways.
- Labeling and manipulation of signaling protein in non-genetic animal models.
