Clear evidence of sleep in invertebrates is still meager. Defined as a distinct state of reduced activity, arousability, attention, and initiative, it is well established in mammals, birds, reptiles, and teleosts. It is commonly defined by additional electroencephalographic criteria that are only well established in mammals and to some extent in birds. Sleep states similar to those in mammals, except for electrical criteria, seem to occur in some invertebrates, based on behavior and some physiological observations. Currently the most compelling evidence for sleep in invertebrates (evidence that meets most standard criteria for sleep) has been obtained in the fruit fly Drosophila melanogaster. However, in mammals, sleep is also characterized by a brain state different from that at rest but awake. The electrophysiological slow wave criterion for this state is not seen in Drosophila or in honey bees. Here, we show that, in crayfish, a behavioral state with elevated threshold for vibratory stimulation is accompanied by a distinctive form of slow wave electrical activity of the brain, quite different from that during waking rest. Therefore, crayfish can attain a sleep state comparable to that of mammals.
Sleep is clearly recognized as a state during which the body, not the brain, is passive. It has been studied mostly in mammals and to some extent in vertebrates (1) whereas studies in invertebrates are few and mostly about behavioral and some electrophysiological aspects (2, 3). However, recent genetic studies in flies (4-6) seem to show that these animals have a clearly distinct sleep state; thus, its demonstration in other invertebrates would show that sleep arose very early in evolution.A common assessment of brain states in mammals is based on the proportions of different frequency segments of the power spectrum of the electroencephalogram (EEG) (labeled alpha, beta, gamma, delta, and theta). To test in an invertebrate for the presence of a brain state that could be equivalent to sleep in vertebrates, we chose crayfish for three main reasons: (i) their relatively large size allows us to chronically implant electrodes on the brain (7); (ii) they are easy to maintain and record under laboratory conditions for extended periods of time, and (iii) we learned to recognize a resting position clearly distinguishable from other positions and during which they seem to be unaware of their environment (8) and had elevated thresholds for arousal.We first videotaped animals during periods of 24 h and noted specific stereotypic body positions, the hours of the day at which these occur, and the length of time spent in each position. At these times, we simultaneously recorded their spontaneous brain electrical activity. We also measured arousal thresholds to mechanical stimulation and ''event-related potentials'' with trains of light flashes that tested expectation, a mildly cognitive function. Because crayfish show several resting states during which they remain motionless, we define ''sleep'' as that condition during which the ...