ORIGINAL RESEARCH article
Front. Neurosci.
Sec. Brain Imaging Methods
Longitudinal Resting-State fMRI of Awake Mice During Habituation: Stress, Head Motion, and Functional Connectivity
Provisionally accepted
- Department of Biomedical Engineering, Sungkyunkwan University College of Science, Suwon-si, Republic of Korea
Select one of your emails
You have multiple emails registered with Frontiers:
Notify me on publication
Please enter your email address:
If you already have an account, please login
You don't have a Frontiers account ? You can register here
Background and purpose: Awake mouse fMRI is a powerful tool for both neuroscience and translational research. To minimize head motion during scanning, habituation under physical restraint is commonly used. However, it remains unclear how stress levels and head motion evolve during habituation, particularly within the MRI environment. Methods: To address this, we repeatedly measured plasma corticosterone (CORT) levels in three groups of mice - controls, mice habituated outside the MRI magnet, and mice habituated within the fMRI environment - and acquired longitudinal resting-state fMRI data daily during an eight-day habituation period and again 15 days post-habituation at 15.2 T. Results: We found that CORT levels initially increased by approximately twofold and gradually decreased during habituation outside the magnet, whereas in mice habituated within the fMRI environment, CORT levels increased two- to fourfold and remained elevated throughout the habituation period. One week after habituation, CORT levels returned to baseline in both groups. Throughout all resting-state fMRI scanning sessions, head motion and functional connectivity remained stable, likely due to the well-designed restraint cradle that permitted paw movement. Conclusion: These results suggest that, for our experimental setup, extending the number of habituation days does not further reduce stress in the MRI environment, provided that head motion remains within acceptable limits.
Keywords: awake mouse fMRI, functional connectivity, habituation, Head motion, Longitudinal fMRI, Resting-state fMRI, stress
Received: 22 Dec 2025; Accepted: 03 Feb 2026.
Copyright: © 2026 Choi, Im and Kim. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Seong-Gi Kim
Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.