Genome-wide association mapping of ethanol sensitivity in the Diversity Outbred mouse population

BackgroundA strong predictor for the development of alcohol use disorders (AUDs) is altered sensitivity to the intoxicating effects of alcohol. Individual differences in the initial sensitivity to alcohol are controlled in part by genetic factors. Mice offer a powerful tool for elucidating the genetic basis of behavioral and physiological traits relevant to AUDs; but conventional experimental crosses have only been able to identify large chromosomal regions rather than specific genes. Genetically diverse, highly recombinant mouse populations allow for the opportunity to observe a wider range of phenotypic variation, offer greater mapping precision, and thus increase the potential for efficient gene identification.MethodsWe have taken advantage of the Diversity Outbred (DO) mouse population to identify and precisely map quantitative trait loci (QTL) associated with ethanol sensitivity. We phenotyped 798 male J:DO mice for three measures of ethanol sensitivity: ataxia, hypothermia, and loss of the righting response. We used high density MEGAMuga and GIGAMuga arrays to obtain genotypes ranging from 77,808 – 143,259 SNPs. In addition, we performed RNA sequencing in striatum to map expression QTLs and to identify gene expression-trait correlations.ResultsWe then applied a systems genetic strategy to identify narrow QTLs and construct the network of correlations that exist between DNA sequence, gene expression values and ethanol-related phenotypes to prioritize our list of positional candidate genes.ConclusionsOur results can be used to identify alleles that contribute to AUDs in humans, elucidate causative biological mechanisms, or assist in the development of novel therapeutic interventions.

The “right” side of sleeping: laterality in resting behaviour of Aldabra giant tortoises (Aldabrachelys gigantea)

Although some studies investigated lateralization in reptiles, little research has been done on chelonians, focusing only on few behaviours such as righting response and escape preference. The aim of this study was to investigate lateralization in Aldabra giant tortoises (Aldabrachelys gigantea), focusing on asymmetrical positioning of the limbs and the head during resting behaviour, called sleep-like behaviour, involving both wild tortoises and individuals under human care. Subjects of the study were 67 adult Aldabra tortoises (54 free ranging on Curieuse, 13 under human care in Mahè Botanical Garden). For each tortoise observed during sleep-like behaviour, we recorded the position of the head (on the left, on the right or in line with the body midline) and we collected which forelimb and hindlimb were kept forward. Moreover, the number of subjects in which limbs were in a symmetrical position during the sleep-like behaviour was recorded. Based on our results, the number of tortoises with asymmetrical position of head and limb was higher (head: 63%; forelimbs: 88%; hindlimbs: 70%) than the number of tortoises with symmetrical position of the head and the limb. Regarding the head, throughout the subjects found with the asymmetrical position of the head during sleep-like behaviour, tortoises positioning the head on the right (42%) were more than those sleeping with the head on the left (21%). We found a relationship between the position of the forelimbs and hindlimbs during sleep-like behaviour. We reported no differences between Mahè (under human care) and Curieuse (wild) tortoises. Findings of this preliminary study underlined traces of group-level lateralization in head positioning during the sleep-like behaviour, possibly due to a left-eye/right-hemisphere involvement in anti-predatory responses and threatening stimuli as reported in reptiles and other vertebrates. This study aims at adding data on brain lateralization, often linked to lateralized behaviours, in reptiles, especially in chelonians.

The costs of being a good dad: egg-carrying and clutch size impair locomotor performance in male midwife toads (Alytes obstetricans)

Parental care is widespread across the animal kingdom. Parental behaviours are beneficial by increasing offspring survival but induce significant costs to the parents. Because parental care is far more common in females, the associated reproductive costs have been largely studied in this sex. Although male parental care is likely to involve significant costs, it has been markedly less well investigated. We studied the costs of egg-carrying on locomotor performance in an amphibian species (Alytes obstetricans) with male parental care. We examined complementary parameters including hopping performance, righting response, hindleg muscle response to egg burden, and homing time in males carrying or not carrying eggs. We found that carrying males showed altered locomotor performance for most traits. In addition, alteration of performance was closely related to relative clutch size. Clutch desertion occurred in smaller individuals carrying larger relative clutch mass, and performance after desertion was similar to that of non-reproductive individuals. Overall, our study demonstrates that carrying eggs significantly alters male mobility and that performance–clutch size trade-offs are relevant in understanding the evolution of paternal care.

Impacts of water temperature on the physiology and behaviours of the sea urchins Heliocidaris crassispina and Mesocentrotus nudus that reflect their range extension and disappearance in the Oga Peninsula, northern Honshu, Japan

Ocean warming has facilitated the extension of Heliocidaris crassispina to Oga Peninsula, Japan, where the native species Mesocentrotus nudus has disappeared. To verify the temperature impacts on the physiology and behaviour of the two species, we reared small sea urchins at the increasing/decreasing temperature rate of 2.5°C week-1. The righting response, lantern reflex, gonad and gut carbon (C) and nitrogen (N) contents, and feeding rate were investigated. The high and low temperature limits of H. crassispina were 33.3°C and 3.9°C, respectively, which were higher than those of M. nudus. The optimal temperature ranges for behaviour and feeding in H. crassispina were 10.3–31.0°C and 10.3–33.4°C, respectively, which were higher than those in M. nudus. Feeding rates decreased significantly in both species when the temperature approached the high or low temperature limit, but the gut C and N contents of were not greatly affected. At 26–31°C, the feeding rate significantly decreased in M. nudus but not in H. crassispina, which may explain the replacement of M. nudus by H. crassispina in the Oga Peninsula.

Multi-stressor Extremes Found on a Tropical Coral Reef Impair Performance

Global change has resulted in oceans that are warmer, more acidic, and lower in oxygen. Individually any one of these stressors can have numerous negative impacts on marine organisms, and in combination they are likely to be particularly detrimental. Understanding the interactions between these factors is important as they often covary, with warming promoting hypoxia, and hypoxia co-occurring with acidification. Few studies have examined how all three factors interact to affect organismal performance, and information is particularly sparse for tropical organisms. Here we documented a strong relationship between high temperatures, low dissolved oxygen (DO), and low pH in and around a tropical bay. We used these field values to inform two multi-stressor experiments. Each experimental factor had two levels, one representing current average conditions and the other representing current extreme conditions experienced in the area. We used sea urchin righting response as a measure of organismal performance for an important reef herbivore. In the first experiment 2-h exposures to a fully factorial combination of temperature, DO, and pH showed that righting success was significantly depressed under low oxygen. To more fully understand the impacts of pH, we acclimated sea urchins to control and low pH for 7 days and subsequently exposed them to the same experimental conditions. Sea urchins acclimated to control pH had significantly reduced righting success compared to animals acclimated to low pH, and righting success was significantly depressed under hypoxia and high temperature, compared to normoxia and ambient temperature. These results show that short, 2 h exposures to the temperature and DO extremes that are already experienced periodically by these animals have measurable detrimental effects on their performance. The positive impact of reduced pH is evident only over longer, 7 days durations, which are not currently experienced in this area.

Effects of Paternal Preconception Vapor Alcohol Exposure Paradigms on Behavioral Responses in Offspring

We and others previously reported that paternal preconception chronic ethanol exposure leads to molecular, physiological, and behavioral changes in offspring including reduced ethanol consumption and preference relative to controls. The goal of the present study was to further explore the impact of paternal ethanol exposure on a wide variety of basal and drug-induced behavioral responses in first generation offspring. Adult male mice were exposed to chronic intermittent vapor ethanol or control conditions for 5–6 weeks before being mated with ethanol-naïve females to produce ethanol (E)- and control (C)-sired offspring. E-sired male offspring showed stress hyporesponsivity in a stress-induced hyperthermia assay and E-sired female offspring had reduced binge-like ethanol consumption in a drinking in the dark assay compared to C-sired offspring. E-sired offspring also showed altered sensitivity to a sedative/hypnotic dose of the GABAergic drug midazolam, but not ketamine or ethanol, in a loss of the righting response assay. E-sired offspring did not differ from controls in marble burying, novel object location, novel object recognition, social interaction, bottle-brush, novelty suppressed feeding, prepulse inhibition, every-other-day ethanol drinking, or home cage activity assays. This study adds to a growing body of literature suggesting that like in utero alcohol exposure, paternal preconception alcohol exposure can also have effects that persist and impact behavior of offspring.

Thermal stress responses of two sympatric crayfishes in Louisiana, Procambarus clarkii Girard, 1852 and Procambarus zonangulus Hobbs & Hobbs, 1990 (Decapoda: Astacoidea: Cambaridae)

Water temperature is an important abiotic component in farmed and wild crayfish habitats as it influences individual and population characteristics including growth, metabolic activity, and overall population vigor. Thermal limits, however, have not been established for the two commercially important crayfishes in Louisiana, the red swamp crayfish Procambarus clarkii (Girard, 1852) and the southern white river crayfish P. zonangulus (Hobbs & Hobbs, 1990). We aimed to determine the thermal stress responses for the two species. Thermal stress was examined by the critical thermal maxima (CTmax) for crayfish acclimated to 24, 27, or 30 °C for two weeks prior to experimentation. Water temperature in all acclimation trials was increased 1 °C h–1 and the end-point temperature for an individual was determined by the failure of righting response within 30 seconds. CTmax values (±SD) for P. clarkii acclimated at 24, 27, and 30 °C were 39.5 ± 0.5, 39.1 ± 0.3, and 39.4 ± 0.7 °C respectively, and 39.8 ± 0.8, 38.8 ± 0.8, and 39.0 ± 0.6 °C for P. zonangulus. CTmax did not differ significantly between species in any of the acclimation treatments. Thermal stress was also examined by measuring P. clarkii and P. zonangulus hemolymph glucose concentration every two hours for crayfish acclimated at 26 °C for two weeks and water temperature increased 1 °C h–1. Hemolymph glucose concentration began to increase at 36 °C in P. zonangulus and 38 °C in P. clarkii. Large increases in mean hemolymph glucose concentration (±SE) were observed in both P. clarkii (67.19 mg–dl ± 24.55) and P. zonangulus (74.11 mg–dl ± 18.04) at 40 °C.

Effects of flow velocity on fitness-related behaviours of the sea urchin Mesocentrotus nudus: new information on stock enhancement

The effects of flow velocity on the fitness-related behaviours of Mesocentrotus nudus remain largely unknown, greatly hampering the efficiency of stock enhancement. To explore the appropriate velocities for stock enhancement, we investigated dislodgement and immobilization velocities up to 90 cm s−1. The experimental results showed that M. nudus (test diameter of ~30 mm) were dislodged at 73.50 ± 7.7 cm s−1 and that M. nudus movement occurred only when the flow velocity was less than 33.40 ± 2.7 cm s−1. Three flow velocities less than 33.40 ± 2.7 cm s−1 (2, 10 and 20 cm s−1) were subsequently used to study the effects of flow velocities on covering behaviour and the righting response time of M. nudus. The downstream movement velocity of M. nudus was significantly larger than that upstream at 2 cm s−1 (P = 0.016) and 10 cm s−1 (P = 0.008), but not at 20 cm s−1 (P = 0.222). The righting response time of M. nudus was significantly longer at 20 cm s−1 than that at 2 cm s−1 (P = 0.015). The present study indicates that a flow velocity less than 20 cm s−1, preferably 2–10 cm s−1, is probably appropriate for the stock enhancement of M. nudus. Notably, the current study is a laboratory investigation without considering the hydrographic complexity in the field. Further studies should be carried out to investigate the long-term effects of water flow on feeding and growth of M. nudus both in the laboratory and the field.

Heat, health and hatchlings: associations of in situ nest temperatures with morphological and physiological characteristics of loggerhead sea turtle hatchlings from Florida

Incubation temperatures, in addition to an embryo’s genetic makeup, are critical in many aspects of adequate sea turtle embryonic development. The effects of high and low incubation temperatures on hatchling quality have been previously examined; however, many of these studies were conducted on relocated or laboratory-reared nests, which do not accurately reflect natural nest temperature fluctuations. To observe the impacts of varying in situ incubation temperatures on loggerhead sea turtle (Caretta caretta) hatchling morphology, various health variables and locomotor performance, temperature data loggers were deployed in 15 loggerhead nests on Juno Beach, Florida, between May and July 2018. Over the course of the study period, 10 morphological traits were measured, blood analytes and heart rate were assessed for the establishment of reference intervals and the self-righting response in seawater was evaluated. Warmer months were associated with smaller body size and higher body condition index, larger umbilical scar size, slower righting time, lower heart rates and higher packed cell volume, hemoglobin, total solids, total white blood cell count, absolute heterophils and absolute basophils. These findings provide evidence that higher in situ incubation temperatures have the potential to adversely affect hatchlings from warmer nests due to increased risk of predation from smaller body sizes, decreased physical responses and overall fitness, altered hemodynamic balance (e.g. dehydration) and potential inflammation and/or stress. With rising temperatures, we predict sea turtle hatchlings may have increasing risks of developing suboptimal physiological features affecting overall fitness and ultimately survival. These results demonstrate that rising environmental temperatures can negatively impact sea turtle hatchlings, thus representing additional stress on sea turtle populations and contributing to our understanding of potential pathophysiological effects of climate change on the delicate life-stage class of the sea turtle hatchling. This information will be useful for formulating effective future sea turtle management plans.