Google map of mouse brain
“The key to success is being in the right place at the right time, recognizing that you are there and taking action!” Ray Kroc
In protein function… it defiantly works the same.
High-resolution large-scale mosaic map. This map provides global views of a whole brain section, and allows identification of subcellular features without losing the context of the brain regions.
Rigorous cellular characterization of protein localization is a necessary step if we aim to understand protein function in a physiological context. To date, relatively few efforts have attempted to systematically map the subcellular localization of endogenous proteins using imaging-based techniques.
We generated high-resolution large-scale mosaic images of whole mouse brain slices using RIβ and RIIβ-specific antibodies. The use of large-scale immunohistochemical brain maps allows us to gain an overview of the RIβ and RIIβ protein distribution over large areas and then zoom in to obtain higher resolution views in order to investigate subcellular features. We found that each regulatory isoform is predominant in distinct brain regions and were able to identify unique and consistent patterns of cellular and subcellular distribution. This information allow us to articulate the functional distinction between PKA isoforms.
Mouse disease models
We are currently utilizing our high resolution maps to study protein localization in disease and therapy and to drive new hypotheses. We use mouse animal models or human brain tissues. They are especially relevant for many neurodegenerative diseases such as Alzheimer’s or Parkinson diseases. They are also relevant for psychiatric illnesses such as mood disorders where PKA signaling is aberrant, or for addiction where PKA plays a prominent role.
Point mutation mouse models using Crispr Cas9 technology
Our lab aims to translate the genomic data into a three dimensional structure to enable a better understanding of the molecular and cellular mechanisms, and then to control it with a specific and precise drug targeting therapy based on a SNP mutation.
Genetically encoded tools correlating light and electron microscopy
To visualize subcellular localization at a nanoscale resolution, we applied a novel chemical biology strategy, called miniSOG (mini Singlet Oxygen Generator), developed by the Nobel Laureate Prof. Roger Tsien and Prof. Mark Ellisman. The miniSOG is a fusable marker that enables to correlated light and electron microscopy to map the localization of a protein. This localization technique provides the highest possible spatial resolution. Here we show that PKA regulatory subunit RIβ is specifically localized to the inner membrane of the mitochondria as well as the nucleus
