Dissecting neuronal-glial cross-talk
Accumulating evidence in the recent decade indicates that the for long neglected glial cells are tightly engaged in brain network function. However, up to now, it is not established how glial activities are modulating neuronal circuit function, especially the field of neuronal-glial communication in awake behaving animals remains unexplored. The lack of tools has hampered progress in glial research. In the present project applied, the applicant aims at developing novel molecular approaches for glial research. The overall objective is to dissect the precise mechanisms by which glia and neurons communicate. Ivermectin (IVM) inducible neuronal silencing, a novel strategy for regional and cell type specific inhibition of neurons, will be used to dissect glial activities contributing to neuronal circuits. The project also utilizes virally delivered genetically encoded fluorescent sensors of various molecules in combination with two-photon microscopy of head-fixed awake mice under behavior tasks.
To minimize the distress of animals and improve animal well-being, I will provide enriched housing conditions, careful handling and caring before, during and after the surgery, and close monitoring during the whole experimental procedure. Suitable anaesthesia and analgesia will be applied during and after surgery.
The unique combination of cutting-edge approaches - the two-photon imaging in acute brain slices and head-fixed behaving mice, genetically encoded sensors, optognetic manipulation (bPAC) and the IVM inducible neuronal silencing - will bring the glial research to a new level. This project will open up a new page of research on neuronal-glial cross-talk to gain a more comprehensive understanding of the workings of the brain.
A total number 220 wild-type C57/bl6J mice, 60 IP3R2 knockout mice, and 30 AQP4KO mice are planned in histology and brain two-photon imaging tasks.
The mouse brain architecture has many similarities to the human brain on the both the macroscopic level andcircuit level. Unfortunately, cell cultures of neurons do not answer questions about how brain cells in the awakebrain are active. We will keep the number of used mice to a minimum. The experience that we have built up in the past years will help us to optimally organize the breeding colonies, and to perform high quality surgeries and image sampling so data can be obtained with fewer mice.
To minimize the distress of animals and improve animal well-being, I will provide enriched housing conditions, careful handling and caring before, during and after the surgery, and close monitoring during the whole experimental procedure. Suitable anaesthesia and analgesia will be applied during and after surgery.
The unique combination of cutting-edge approaches - the two-photon imaging in acute brain slices and head-fixed behaving mice, genetically encoded sensors, optognetic manipulation (bPAC) and the IVM inducible neuronal silencing - will bring the glial research to a new level. This project will open up a new page of research on neuronal-glial cross-talk to gain a more comprehensive understanding of the workings of the brain.
A total number 220 wild-type C57/bl6J mice, 60 IP3R2 knockout mice, and 30 AQP4KO mice are planned in histology and brain two-photon imaging tasks.
The mouse brain architecture has many similarities to the human brain on the both the macroscopic level andcircuit level. Unfortunately, cell cultures of neurons do not answer questions about how brain cells in the awakebrain are active. We will keep the number of used mice to a minimum. The experience that we have built up in the past years will help us to optimally organize the breeding colonies, and to perform high quality surgeries and image sampling so data can be obtained with fewer mice.