JACUZI-SD: An automated, high-throughput, minimally stressful approach to sleep depriving larval zebrafish

Inadequate sleep is common and broadly disrupts well-being. A fundamental challenge to modeling sleep deprivation (SD) experimentally is the introduction of confounds such as stress or circadian manipulation. We therefore developed JACUZI-SD (jetting automated currents under zebrafish to induce sleep deprivation), an automated, high-throughput, and minimally stressful system to SD larval zebrafish during the dark cycle. JACUZI-SD delivers randomized water pulses via a custom milli-fluidic device integrated with a 96-well plate, reducing total sleep time by 41%-64% and eliciting a rebound sleep characterized by an 11% increase in total sleep time and 29% increase in sleep bout length. Water pulses are minimally stressful, eliciting less fosab expression in the hypothalamic-pituitary-interrenal (HPI) stress axis compared to other SD methods. Additionally, we observe galanin induction, an established response to heightened sleep pressure. JACUZI-SD provides a powerful, minimally invasive platform for dissecting the neural underpinnings and biological variability of sleep homeostasis in vertebrates.