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The investigation of neural circuits is important for interpreting both healthy brain function and psychiatric disorders. Currently, the architecture of neural circuits is always investigated with fluorescent protein encoding neurotropic virus and ex vivo fluorescent imaging technology. However, it is difficult to obtain a whole‐brain neural circuit connection in living animals, due to the limited fluorescent imaging depth. Herein, the noninvasive, whole‐brain imaging technique of MRI and the hypotoxicity virus vector AAV (adeno‐associated virus) were combined to investigate the whole‐brain neural circuits in vivo. AAV2‐retro are an artificially‐evolved virus vector that permits access to the terminal of neurons and retrograde transport to their cell bodies. By expressing the ferritin protein which could accumulate iron ions and influence the MRI contrast, the neurotropic virus can cause MRI signal changes in the infected regions. For mice injected with the ferritin‐encoding virus vector (rAAV2‐retro‐CAG‐Ferritin) in the caudate putamen (CPu), several regions showed significant changes in MRI contrasts, such as PFC (prefrontal cortex), HIP (hippocampus), Ins (insular cortex) and BLA (basolateral amygdala). The expression of ferritin in those regions was also verified with ex vivo fluorescence imaging. In addition, we demonstrated that changes in T2 relaxation time could be used to identify the spread area of the virus in the brain over time. Thus, the neural connections could be longitudinally detected with the in vivo MRI method. This novel technique could be utilized to observe the viral infection process and detect the neural circuits in a living animal.

Longitudinal neural connection detection using a ferritin‐encoding adeno‐associated virus vector and in vivo MRI method