Sato, D. X.; Chatzigiannis, M. M.; Shoji, H.; Sala, G.; Hattori, S.; Takao, K.; Kinashi, T.; Kishino, T.; Morita, S.; Hatada, I.; Shinoda, Y.; Hattori, K.; Yagi, T.; Matsumoto, A.; Egawa, H.; Nishihara, S.; Shimizu, K.; Ikegami, K.; Yamada, M. K.; Ageta, H.; Setou, M.; Tao, H.; Ueno, N.; Bhandari, P.; Shigemoto, R.; Wakatsuki, S.; Araki, T.; Yamanaka, A.; Mukai, H.; Nagaoka, T.; Kishi, M.; Furuya, S.; Yamamoto, T.; Kubo, Y.; Iida, Y.; Kazuki, Y.; Enomoto, H.; Fukasawa, M.; Usuda, N.; Inoue, S.; Inokuchi, K.; Hattori, T.; Taniguchi-Ikeda, M.; Toda, T.; Kubo, A.; Kawaai, K.; Mikoshiba, K.; van d

Fear conditioning is widely used to assess associative memory in mice, yet percent freezing conflates memory with baseline locomotor and anxiety-related traits. A systematic survey of recent studies (2020-2025) found that fewer than 1% statistically integrate locomotor activity into freezing analyses. Here, we address this gap using a large-scale dataset of >10,000 mice across >160 comparisons, including genetic mutations, pharmacological interventions and aging, tested in 15 standardized behavioral paradigms. Conventional freezing scores covaried strongly with general locomotor activity, obscuring memory-related phenotypes. Multiple factor analysis identified two principal behavioral dimensions, locomotor activity and learning/memory: conventional freezing aligned with the locomotor dimension, whereas freezing subtraction and the activity suppression ratio mapped onto the memory dimension and improved detection of synaptic plasticity phenotypes. These analyses show that baseline locomotor normalization is essential for interpreting fear conditioning as a memory assay and provide an open framework for selecting and reporting locomotor-normalized metrics.