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. 2024 Dec;341(10):1121-1129.
doi: 10.1002/jez.2846. Epub 2024 Jul 3.

An electrophysiological correlate of sleep in a shark

Affiliations

An electrophysiological correlate of sleep in a shark

John A Lesku et al. J Exp Zool A Ecol Integr Physiol. 2024 Dec.

Abstract

Sleep is a prominent physiological state observed across the animal kingdom. Yet, for some animals, our ability to identify sleep can be masked by behaviors otherwise associated with being awake, such as for some sharks that must swim continuously to push oxygenated seawater over their gills to breathe. We know that sleep in buccal pumping sharks with clear rest/activity cycles, such as draughtsboard sharks (Cephaloscyllium isabellum, Bonnaterre, 1788), manifests as a behavioral shutdown, postural relaxation, reduced responsiveness, and a lowered metabolic rate. However, these features of sleep do not lend themselves well to animals that swim nonstop. In addition to video and accelerometry recordings, we tried to explore the electrophysiological correlates of sleep in draughtsboard sharks using electroencephalography (EEG), electromyography, and electrooculography, while monitoring brain temperature. The seven channels of EEG activity had a surprising level of (apparent) instability when animals were swimming, but also when sleeping. The amount of stable EEG signals was too low for replication within- and across individuals. Eye movements were not measurable, owing to instability of the reference electrode. Based on an established behavioral characterization of sleep in draughtsboard sharks, we offer the original finding that muscle tone was strongest during active wakefulness, lower in quietly awake sharks, and lowest in sleeping sharks. We also offer several critical suggestions on how to improve techniques for characterizing sleep electrophysiology in future studies on elasmobranchs, particularly for those that swim continuously. Ultimately, these approaches will provide important insights into the evolutionary confluence of behaviors typically associated with wakefulness and sleep.

Keywords: biologging; cartilaginous fish; elasmobranch; electroencephalogram; electromyography; electrooculogram; sleep evolution.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
(a) Top‐down view of the acrylic stereotaxic holder. The flattened white tube near the top of the panel delivered buffered MS‐222‐infused seawater into the buccal cavity and out over the gills; the hole at the other end of the surgery tank housed an adjustable pipe for overflow drainage to the sump (when fitted). (b) An anesthetized draughtsboard shark secured in the stereotaxic holder by “ear bars” and nose clamp. (c) At the end of the surgery, the surgical site was closed and the data logger sutured in place. (d) Animals were filmed from above; they appeared as black silhouettes owing to the diffuse, near‐infrared lighting beneath.
Figure 2
Figure 2
Segmented images from computed tomography (CT) scan for two of the three experimental draughtsboard shark brains from lateral (top row) and dorsal (bottom row) viewpoints. Imaging of the third shark was not successful. Shown are referential electrodes for the telencephalon, and cerebellum or optic tectum (green), reference screws (blue), thermistor (red), and anchor screws (yellow). In all images, the brainstem is on the left. Owing to tissue shrinkage during fixation, it is possible that electrodes seen above the brain here, were touching the brain in vivo. Cb, cerebellum; D, diencephalon; Md, medulla oblongata; Ot, optic tectum; Tel, telencephalon.
Figure 3
Figure 3
Exemplar raw‐data electrophysiological traces from a shark actively awake (green), quietly awake (blue), and asleep (red). Signals reflect our attempt to record skeletomuscular tone (electromyogram; high‐pass filter: 10 Hz, scale: 40 µV), movement (accelerometry in X, Y, and Z axes; scale: 1 G), ocular activity (electrooculogram; low‐pass filter: 10 Hz, scale: 50 µV) followed by signals from the left (L) or right (R) anterior (A) or posterior (P) telencephalon, cerebellum, and diencephalon (all scaled at 20 µV). Using the computed tomography (CT) scans, it appears as though the diencephalon and telencephalon electrodes were in contact with the surface of the brain, but the RP electrode penetrated the dura. Both cerebellar electrodes (unintentionally) also penetrated the dura. These traces show that brain activity was obscured in swimming sharks. Yet, even in stationary sharks, signals were unstable. X‐axis scale bars denote 5 s, such that each trace is 20 s.
Figure 4
Figure 4
Processed accelerometry and electromyogram signals over a representative 24‐h day from one shark housed under a 12:12 light:dark photoperiod. Gray shading denotes the 12‐h night, starting at 1900 h. These plots show the strong alignment between movement and muscle tone during active (green) and quiet (blue) wakefulness, and sleep (red).
Figure 5
Figure 5
The degree to which sharks moved (top) and their level of muscle tone (bottom) depended on whether they were actively or quietly awake, or asleep. Muscle tone was highest during active wakefulness and lowest during sleep. Datapoints represent 24‐h means per shark, with 1–6 days per shark. Significance is marked by asterisks (p = 0.03*; p < 0.001***).

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