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Genetically targeted reporter imaging of deep neuronal network in the mammalian brain

Shimojo, M.; Ono, M.; Takuwa, H.; Mimura, K.; Nagai, Y.; Fujinaga, M.; Kikuchi, T.; Okada, M.; Seki, C.; Tokunaga, M.; Maeda, J.; Takado, Y.; Takahashi, M.; Minamihisamatsu, T.; Zhang, M.-R.; Tomita, Y.; Suzuki, N.; Maximov, A.; Suhara, T.; Minamimoto, T.; Sahara, N.; Higuchi, M.

2020-04-09 neuroscience
10.1101/2020.04.08.032870 bioRxiv
Show abstract

Positron Emission Tomography (PET) allows biomolecular tracking, while PET monitoring of brain networks has been hampered by the lack of a suitable reporter. Here, we describe in vivo brain imaging that takes advantage of bacterial dihydrofolate reductase, ecDHFR, and its unique antagonist, TMP. In mice, peripheral administration of radiofluorinated and fluorescent TMP analogs enabled PET and intravital microscopy, respectively, of neuronal ecDHFR expressions. This technique is applicable to the visualization of neuronal ensemble activities elicited by chemogenetic manipulation in the mouse hippocampus. Notably, ecDHFR-PET offers mapping of neuronal projections in non-human primate brains, indicating the availability of ecDHFR-based tracking technologies for network monitoring. Finally, we demonstrate the utility of TMP analogs for PET assays of turnover and self-assembly of proteins tagged with ecDHFR mutants. Our findings may facilitate a broad spectrum of PET analyses of a mammalian brain circuit at molecular levels that were not previously applicable for technical reasons.

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