Time-course profiling of behavioral phenotypes and Ywhah/ERK signaling expression within the vestibular region of the brainstem during motion sickness habituation.

Motion sickness habituation represents a core neuroplastic model for investigating vestibular disorder adaptation, yet its brain-region-specific temporal molecular mechanisms remain poorly defined. Here, we adopted multi-brain-region proteomic screening combined with time-series molecular validation to characterize key molecular dynamics during motion sickness habituation. Adult mice received 15 consecutive days of sinusoidal rotational stimulation, and model establishment was verified via four behavioral indices. A self-constructed five-dimensional weighted scoring system based on proteomic data confirmed that the vestibular region of the brainstem exhibited the most prominent molecular response among the hippocampus, cerebral cortex, and brainstem vestibular region. Candidate molecules were further screened by integrating network centrality, statistical significance, functional relevance, druggability, and neurological disease correlation. Reverse transcription quantitative real-time PCR and Western blot were performed to detect the expression changes of candidate proteins in brainstem vestibular region tissues of the complete time-series restraint-only control group and habituation group. Temporal validation revealed progressive downregulation of Ywhah and a time-dependent increase in the ratio of phosphorylated Erk1/2 to total Erk1/2 in the vestibular region of the brainstem. The opposite expression trends of Ywhah and p-Erk1/2/total Erk1/2 first reached statistical significance on Day 5, indicating temporal coupling. In summary, this study delineates distinct temporal molecular patterns in the brainstem vestibular region during motion sickness habituation, with overlapping critical time windows for Ywhah suppression and ERK activation. These findings provide reliable longitudinal molecular evidence and promising candidate targets for further exploring the neuroplastic mechanisms of vestibular habituation.