To verify amylase's connection to sleep loss, Shaw's lab monitored its activity level after sleep deprivation in different fruit fly lines genetically altered to modify their sleep drive.
In one key test, amylase did not increase in a fly modified to endure sleep deprivation longer than normal flies without incurring sleep debt. When scientists kept the same mutant flies awake for extended nine or 12 hour stretches that normally cause them to incur sleep debt, their amylase levels increased.
"This helped prove that the increases in amylase activity level we were seeing weren't just triggered by wakefulness," Shaw says.
Humans kept awake for 28 hours also had increased amylase levels versus controls allowed to sleep normally.
Shaw's lab previously showed that they can use caffeine and methamphetamine to keep flies awake. Caffeine inflicts sleep debt, causing flies to sleep for extended periods when it wears off, while methamphetamine does not. When they monitored fly amylase levels in response to these drugs, they found caffeine drove amylase activity up while methamphetamine did not.
Flies dosed with the herbicide paraquat did not have increased amylase levels, suggesting changes in amylase activity were not related to stress. Flies lacking the gene for amylase had normal sleep and waking cycles, showing that while amylase is tightly linked to sleep drive, it is not actively involved in its regulation.
"We're very pleased with how tightly amylase levels correlate with sleep debt, but for a good diagnostic test we're likely going to need more than one biomarker," Shaw says. "So we're going to continue to use the processes that we've developed to look for other substances that change in connection with the level of sleep debt."
Stephen L. Duntley, M.D., associate professor
Source:Washington University School of Medicine