Simultaneous voltage and calcium imaging and mRNA extraction from single neurons in-vivo
Okinawa Institute of Science and Technology Graduate University (OIST) Staff Scientist
Neurons are the basic building blocks of the nervous system. Understanding their function in the living animal is essential for understanding the nervous system and curing neurological diseases. For a deeper understanding of neuronal function, it is necessary to record multidimensionally in the awake behaving animal. Additionally, to connect form with function, it is essential to reconstruct their complete 3D structure and to determine their protein expression pattern. In our previous work I was able to record functional activity by imaging voltage and calcium signals in cerebellar Purkinje neurons in awake animals with unprecedented spatial and temporal resolution. I have also developed a technique which allows access to neurons in vivo, for electrophysiology, pharmacological manipulation and/or mRNA extraction. Together, this allows us to acquire functional and structural data from Purkinje neurons and afterwards extract mRNA from their soma. I will provide collaborators with mRNA samples to solve the following problem: Purkinje neurons have previously been considered a homogeneous cell type. The expression pattern of a few proteins and their complex signaling patterns, however, tell us this is not the case and the relationship between their unique protein expression and complex signaling patterns remains undetermined. By combining functional recording and mRNA screening techniques it will be possible to identify the different types of Purkinje neurons based on their functional activity and protein expression patterns.
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- Roome CJ, Kuhn B, “Voltage imaging with ANNINE dyes and two-photon microscopy of Purkinje dendrites in awake mice”, Neuroscience Research. 2019, Nov 20.