Altered Rest-Activity Patterns Evolve via Circadian Independent Mechanisms in Cave Adapted Balitorid Loaches

Circadian rhythms and rest homeostasis are independent processes, each regulating important components of rest-activity patterns. Evolutionarily, the two are distinct from one another; total rest time is maintained unaffected even when circadian pacemaker cells are ablated. Throughout the animal kingdom, there exists a huge variation in rest-activity patterns, yet it is unclear how these behaviors have evolved. Here we show that four species of balitorid cavefish have greatly reduced rest times in comparison to rest times of their surface relatives. All four cave species retained biological rhythmicity, and in three of the four there is a pronounced 24-hour rhythm; in the fourth there is an altered rhythmicity of 38–40 hours. Thus, consistent changes in total rest have evolved in these species independent of circadian rhythmicity. Taken together, our data suggest that consistent reduction in total rest times were accomplished evolutionarily through alterations in rest homeostasis

Distinct genetic architecture underlies the emergence of sleep loss and prey-seeking behavior in the Mexican cavefish

Sleep is characterized by extended periods of quiescence and reduced responsiveness to sensory stimuli. Animals ranging from insects to mammals adapt to environments with limited food by suppressing sleep and enhancing their response to food cues, yet little is known about the genetic and evolutionary relationship between these processes. The blind Mexican cavefish, Astyanax mexicanus is a powerful model for elucidating the genetic mechanisms underlying behavioral evolution. A. mexicanus comprises an extant ancestral-type surface dwelling morph and at least five independently evolved cave populations. Evolutionary convergence on sleep loss and vibration attraction behavior, which is involved in prey seeking, have been documented in cavefish raising the possibility that enhanced sensory responsiveness underlies changes in sleep. We established a system to study sleep and vibration attraction behavior in adult A. mexicanus and used high coverage quantitative trait loci (QTL) mapping to investigate the functional and evolutionary relationship between these traits. Analysis of surface-cave F2 hybrid fish and an outbred cave population indicates that independent genetic factors underlie changes in sleep/locomotor activity and vibration attraction behavior. High-coverage QTL mapping with genotyping-by-sequencing technology identify two novel QTL intervals that associate with locomotor activity and include the narcolepsy-associated tp53 regulating kinase. These QTLs represent the first genomic localization of locomotor activity in cavefish and are distinct from two QTLs previously identified as associating with vibration attraction behavior. Taken together, these results localize genomic regions underlying sleep/locomotor and sensory changes in cavefish populations and provide evidence that sleep loss evolved independently from enhanced sensory responsiveness.https://doi.org/10.1186/s12915-015-0119-

Reduced total rest time in cave balitorids is primarily a function of reduced rest bout duration.

<p>Quantification of total rest (A.) indicates that surface fish spend an average of 999.1±64.4 min per 24 hr in a rest-like state. In strong contrast to the surface dwelling form, average total rest per 24-hour period was significantly less in all four of the cave species studied (<i>S. oedipus</i>: 128.8±62.04 min, p<0.001; <i>S. jaruthanini</i>: 130.4±27.38 min, p<0.001; <i>N. troglocataractus</i>: 202.3±122.1 min, p<0.001; <i>S. speisi</i>: 161.0±72.51 min, p<0.001). (B–C.) There were no significant differences between the number of rest bouts in surface and cave balitorids (B. Day - Epigean: 4.4±1.3; S. oedipus: 14.2±5.5; S. jaruthanini: 5.4±1.5; N. <i>troglocataractus</i>: 43.0±32.1; S. speisi: 10.7±3.5; Night - Epigean: 4.4±1.4; S. oedipus: 17.1±8.2; S. jaruthanini: 14.0±2.8; N. <i>troglocataractus</i>: 22.0±12.3; S. speisi: 47.3±29.3) thought the duration of each rest bout was significantly less in all cave populations studied compared to surface (Day - Epigean: 185.8±71.7; <i>S. oedipus</i>: 13.2±0.9, p<0.001; <i>S. jaruthanini</i>: 16.8±1.8, p<0.001; <i>N. troglocataractus</i>: 14.4±1.4, p<0.001; <i>S. speisi</i>: 12.7±0.6, p<0.001; Night - Epigean: 219.8±50.7; S. oedipus: 15.2±1.3, p<0.001; <i>S. jaruthanini</i>: 16.7±0.7, p<0.001; <i>N. troglocataractus</i>: 15.4±3.1, p<0.001; <i>S. speisi</i>: 12.5±0.1, p<0.001). For panel A., white denotes subjective day (CT0–CT12) and black denoted subjective night (CT12–CT24). For panels B. and C., Black denotes surface, dark-shade grey denotes <i>S. jaruthanini</i>, mid-shade grey denotes <i>S. spesei</i>, light-shade grey denotes <i>N. troglocataractus</i> and white denotes <i>S. oedipus</i>. All plots represent mean ± standard error of the mean (SEM). Asterisks represent significance relative to surface fish.</p

A protocol for whole-brain Ca2+ imaging in Astyanax mexicanus, a model of comparative evolution

Summary: In this protocol, we describe a comparative approach to study the evolution of brain function in the Mexican tetra, Astyanax mexicanus. We developed surface fish and two independent populations of cavefish with pan-neuronal expression of the Ca2+ sensor GCaMP6s. We describe a methodology to prepare samples and image activity across the optic tectum and olfactory bulb. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics

Changes in local interaction rules during ontogeny underlie the evolution of collective behavior

Summary: Collective motion emerges from individual interactions which produce group-wide patterns in behavior. While adaptive changes to collective motion are observed across animal species, how local interactions change when these collective behaviors evolve is poorly understood. Here, we use the Mexican tetra, Astyanax mexicanus, which exists as a schooling surface form and a non-schooling cave form, to study differences in how fish alter their swimming in response to neighbors across ontogeny and between evolutionarily diverged populations. We find that surface fish undergo a transition to schooling mediated by changes in the way fish modulate speed and turning relative to neighbors. This transition begins with the tendency to align to neighbors emerging by 28 days post-fertilization and ends with the emergence of robust attraction by 70 days post-fertilization. Cavefish exhibit neither alignment nor attraction at any stage of development. These results reveal how evolution alters local interactions to produce striking differences in collective behavior