This method can help answer key questions in the sleep field such as the affects of chronic sleep restriction during postnatal development. The main advantage of this technique is that it is less stressful than other sleep deprivation techniques and can easily be used throughout development and continue into adulthood. Begin by assigning each cage to either the sleep deprivation or control group.
Ensure that mice have access to food and water ad libitum, and remove any food, bedding, or enrichment objects that might restrict the view of the mice. For the control group, remove the sire from the breeder cage on postnatal day five and leave the dam with the pups until weaning. Next, move mice assigned to the control group to the sleep deprivation room and gently prod the mice with a paintbrush continuously for 10 minutes a day for the duration of the experiment.
After 10 minutes, return the mice to the animal holding room and do not disturb them. Begin by removing the sire from the breeder cage on postnatal day five and leave the dam with the pups until weaning. Move mice assigned to the sleep deprivation group to the sleep deprivation room daily during the light cycle where they will be monitored.
During this time, gently prod the mouse with a paintbrush if suspected of being asleep until a response is observed. Alternatively, invert mice and push over onto their backs to disrupt sleep. Consider a mouse to be asleep if any of the following occur:twitching, inactivity, or if eyes are closed.
Consider mice to be awake if they are moving around, trying to flip over after being on their backs, or grooming themselves. If the dam is covering the pups during the sleep deprivation, gently prod her away from the pups so the view of the pups is unrestricted. Continue with sleep deprivation protocol until the experiment endpoint.
When the three-hour period of sleep deprivation is complete, put the bedding and any food that may have been removed back and put the lid back on the cage. Finally, return cages to the animal holding room until the next day of sleep deprivation. Following a one month recovery period, results indicated that in the sociability test, sleep restricted females spent more time sniffing the stranger mouse than control females and sleep restricted males had a tendency to spend more time sniffing the object when compared to control males.
Further, in the social novelty phase of testing, sleep restricted female mice spent an increased amount of time sniffing the novel mouse than female controls. While attempting this procedure, it's important to remember to pay attention and promptly respond to pre-sleep behaviors, especially in neonatal mice where sleep need is high. Following this procedure, other methods like open field, rotarod, elevated plus maze, and social behavior testing can be performed in order to answer additional questions like the long-term behavioral affects of chronic sleep restriction during development.
