Do twitching and twitch-spindle coupling differ between N2 and N3 sleep in 6-month-olds?

Abstract Twitches are discrete movements that characterize REM sleep. However, recent work showed that twitches also occur during NREM sleep in human infants beginning around 3 months of age, a time when sleep spindles and the cortical delta rhythm are also emerging. Further, NREM twitches are coupled with sleep spindles, suggesting a unique contribution to sensorimotor development. Given that NREM sleep is composed of distinct substages, we investigated whether twitching and twitch-spindle coupling are differentially expressed during N2 and N3 sleep. In 6-month-old human infants (n=21; 7 females), we recorded EEG, respiration, and video during daytime sleep. We found high-intensity twitching during N2 and REM but not N3 sleep. In contrast, sleep spindles exhibited similar temporal characteristics during N2 and N3. Also, despite differences in the intensity of twitching during N2 and N3, significant twitch-spindle coupling occurred in both stages. Finally, the rate of twitching was inversely related to delta power across NREM periods. These findings suggest that although twitching occurs during REM, N2, and N3 sleep at this age, its expression is compatible with some sleep components (e.g., rapid eye movements, sleep spindles) but not others (e.g., cortical delta), highlighting the continuing need to better understand the dynamic organization of sleep and its individual components in early development. Statement of significance Twitching has long been recognized as a core component of mammalian REM sleep. However, twitching also occurs during non-REM (NREM) sleep in human infants beginning around 3 months of age. Given that NREM sleep is composed of distinct stages (including N2 and N3), we asked whether the expression of twitching and its temporal coupling with sleep spindles is different between the two stages. Even though twitching was more intense during N2 than N3, twitch-spindle coupling occurred in both stages, again suggesting a unique and likely transient functional contribution to sensorimotor development. A necessary next step is to delineate the developmental trajectory of twitching and twitch-spindle coupling beyond 6 months of age. Competing Interest Statement The authors have declared no competing interest. Footnotes This manuscript was revised and resubmitted after peer review.