Leatherback turtles: the menace of plastic.

The leatherback, Dermochelyscoriacea, is a large sea turtle that feeds primarily on jellyfish. Floating plastic garbage could be mistaken for such prey. Autopsy records of 408 leatherback turtles, spanning 123 years (1885-2007), were studied for the presence or absence of plastic in the GI tract. Plastic was reported in 34% of these cases. If only cases from our first report (1968) of plastic were considered, the figure was 37%. Blockage of the gut by plastic was mentioned in some accounts. These findings are discussed in the context of removal of top predators from poorly understood food chains.

Divergent phenotypes of vision and accessory visual function in mice with visual cycle dysfunction (Rpe65 rd12) or retinal degeneration (rd/rd).

PURPOSE: Tests of vision for mice remain limited and the visual phenotype of some retinal disorders in mice remain poorly understood. A novel assay of vision was used to determine how the form and extent of retinal disease affects visual phenotype in mice. METHODS: Retinal histology, the suppression of locomotion by light and visual guidance of locomotion, were assessed in mice with progressive photoreceptor degeneration (rd/rd) or visual cycle dysfunction (Rpe65 rd12). RESULTS: In wild-type mice, there was visual guidance of locomotor activity in dim light and suppression of activity (negative masking) in bright light. In rd/rd mice, vision was sufficient to guide locomotion at postnatal day (P)34 but was lost from P46 onward. In bright light rd/rd mice had enhanced negative masking. Although Rpe65 rd12 mice had no dim light response, with high illumination, vision was sufficient to guide locomotion at all ages tested. CONCLUSIONS: A major concern for gene and cell replacement therapies is the development of visual pathways through which restored retinal function can connect to visual centers of the brain. The residual retinal response to high illumination in Rpe65 rd12 mice translates into useful vision, and visual pathways remain functional--a prerequisite for restoring vision in disorders of the retina. Similarly, useful vision in young rd/rd mice shows that there is visual pathway function before photoreceptor degeneration and suggests the potential for early therapy. Together, these findings recommend observation of masking responses in the assessment of gene and cell replacement therapies.

Negative and positive masking responses to light in retinal degenerate slow (rds/rds) mice during aging.

Bright light suppresses locomotor activity in mice (negative masking) but dim light augments activity (positive masking). Retinal degeneration slow mice (rds/rds) were tested for responses to light at 3 months, 1 and 2 years old. The suppressive effect of light increased between 1 and 2 years, but the positive response to dim lights was severely reduced at 1 year. No such effects occurred in aging wildtypes. The results indicate that enhancement of negative masking depends on the degree of degeneration of the classical photoreceptors, and that residual function in photoreceptors lacking outer segments is initially sufficient for positive masking.

Classical and melanopsin photoreception in irradiance detection: negative masking of locomotor activity by light.

Studies in mice lacking either classical or melanopsin photoreception have been useful in describing the photoreceptor contribution to irradiance detection in accessory visual responses. However, application of these findings to irradiance detection in intact animals is problematical because retinal degeneration or manipulation can induce secondary changes in the retina. Among responses dependent on irradiance detection, the suppression of activity by light (negative masking) has had limited study. To further understand the function of classical and melanopsin photoreceptors we studied irradiance and spectral sensitivity of masking by light, primarily in mice with intact retinae. The sensitivity of negative masking was equivalent for medium ( approximately 500 nm) and short wavelengths ( approximately 365 nm) in three strains of wild-type mice, identifying a marked short-wavelength-sensitive-cone input. At medium wavelengths, spectral sensitivity above 500 nm had closest fit to the nomogram for the medium-wavelength-sensitive-cone, but a combined input of cone and melanopsin photoreceptors in wild-type mice seems likely. Under white light a decompression of the irradiance range of masking in C3H rd/rd cl mice, lacking rods and cones, identified a functional deficiency presumably resulting from the absence of classical photoreceptor input. Together the evidence demonstrates a pronounced and sustained classical photoreceptor input to irradiance detection for negative masking, and suggests one role of classical photoreceptor input is to constrain dynamic range.

Non-photic phase resetting of Dexras1 deficient mice: a more complicated story.

Recently, it has been reported that mice deficient for Dexras1 have a diminished phase-shifting response to photic stimuli but an enhanced response to non-photic stimuli; the latter is of additional interest in that mice generally show relatively weak and unreliable responses to non-photic events. Therefore, in situations in which both photic and non-photic stimuli are present, control of circadian rhythms, relative to wild-types, should tip toward non-photic stimuli in Dexras1(-/-) mice. However, we detected no differences in an experiment in which photic and non-photic entraining agents were presented 180 degrees out of phase, i.e. were in conflict with each other. Furthermore, Dexras1(-/-) and wild-type mice did not differ in non-photic phase shifting to a pulse of confinement in a novel running wheel. Suppression of locomotion by light (masking effect) did not differ between the genotypes, indicating that the photoreceptor input to the non-image forming system is intact. The circadian phenotype of Dexras1(-/-) mice appears to be more complicated than previously thought.

Inter-seasonal maintenance of individual nest site preferences in hawksbill sea turtles.

Within a single population of hawksbill sea turtles (Eretmochelys imbricata), we found a behavioral polymorphism for maternal nest site choice with respect to beach microhabitat characteristics. Some females preferred to nest in littoral forest and in places with overstory vegetation cover, and others preferred to nest in more open, deforested areas. Nest site choice was consistent within and between nesting seasons two years apart. This was not a result of females simply returning to the same location along the shoreline; beach sections used by individual turtles varied between seasons. Nest site choice was not influenced by changes in beach environment (e.g., beach width and foliage cover) or by changes in females' reproductive output (e.g., clutch size), suggesting that fidelity to particular microhabitats is a major determinant of the observed nesting patterns. Because hawksbills exhibit temperature-dependent sex determination, if the behavioral polymorphism in nest site choice has a genetic basis, as is plausible, then this would have implications for sex ratio evolution and offspring survival. By taking an individual-based approach to the study of maternal behavior we reveal previously overlooked individual variation and hope to provide some impetus for more detailed studies of nest site choice.

Distorting gene pools by conservation: Assessing the case of doomed turtle eggs.

Sea turtles have a high reproductive output and high mortality at early stages of the life cycle. In particular, many nests are laid below or close to high tide lines, and subsequently large numbers of eggs may be inundated and destroyed. A common conservation procedure is to relocate such doomed eggs to higher ground. This article examines this practice in the light of recent data revealing that some individual turtles tend to nest relatively near the water and others relatively higher up the beach. Discussion is focused on the question of why apparently poor placement of nests has not been selected against. Comparison between the ecology of leatherback and hawksbill turtle nesting beaches suggests that predictability of environmental conditions on the nesting beaches has an important influence on patterns of nest-site selection. Options are outlined for the management of nesting beaches where a high proportion of turtle eggs is subject to destruction by flooding.

Deforestation: risk of sex ratio distortion in hawksbill sea turtles.

Phenotypic sex in sea turtles is determined by nest incubation temperatures, with warmer temperatures producing females and cooler temperatures producing males. The common finding of highly skewed female-biased hatchling sex ratios in sea turtle populations could have serious repercussions for the long-term survival of these species and prompted us to examine the thermal profile of a relatively pristine hawksbill nesting beach in Guadeloupe, French West Indies. Data loggers placed at nest depth revealed that temperatures in the forested areas were significantly cooler than temperatures in the more open, deforested areas. Using these temperatures as a predictor of sex ratio, we were able to assess the relative contributions of the different beach zones to the primary sex ratio: significantly more males were likely to be produced in the forested areas. Coastal forests are therefore important male-producing areas for the hawksbill sea turtle, and this has urgent conservation implications. On Guadeloupe, as on many Caribbean islands, deforestation rates are high and show few signs of slowing, as there is continual pressure to develop beachfront areas. The destruction of coastal forest could have serious consequences both in terms of local nesting behavior and of regional demography through the effects on population sex ratios. Human alterations to nesting habitat in other reptile taxa have been shown to modify the thermal properties of nest sites in ways that can disrupt their ecology by allowing parasite transmission, increasing vulnerability to climate change, or rendering existing habitat unsuitable.

Scheduled wheel access during daytime: A method for studying conflicting zeitgebers.

It is often stated that light is the primary environmental cue (zeitgeber) for entrainment of circadian clocks. Here, we use a new conflict test design in Syrian hamsters comparing the strength of a photic zeitgeber to that of a non-photic cue, i.e. wheel availability. Re-entrainment to an inverted LD cycle was significantly slowed down in the nocturnal hamster by restricting wheel access to the light phase of the inverted LD cycle. This effect is more pronounced if the illuminance level of the entraining lights is 0.1 lx compared to 6 lx. In this conflict design, the hamsters did not re-entrain to an inverted LD cycle for up to four weeks (when the experiment ended), but voluntarily ran during the light phase. This approximates the situation in people subjected to shift work or jet lag.

Diurnal mice (Mus musculus) and other examples of temporal niche switching.

Examples are presented of nocturnal animals becoming diurnal or vice versa as a result of mutations, genetic manipulations, or brain lesions. Understanding these cases could give insight into mechanisms employed when switches of temporal niche occur as part of the life cycle, or in response to circumstances such as availability of food. A two-process account of niche switching is advocated, involving both a change in clock-controlled outputs and a change in the direct response to light (i.e. masking). An emerging theme from this review is the suggestion that retinal inputs have a greater role in switching than suspected previously.

Masking in waved-2 mice: EGF receptor control of locomotion questioned.

It has been suggested that epidermal growth factors (EGF) are responsible for the inhibition of locomotion by light (i.e., masking) in nocturnal rodents (Kramer et al., 2001). The poor masking response of waved-2 (Egfr(wa2)) mutant mice, with reduced EGF receptor activity, was adduced in support of this idea. In the present work, we studied the responses to light over a large range in illumination levels, in a variety of tests, with pulses of light and with ultradian light-dark cycles in Egfr(wa2) mutant mice. No evidence suggested that normal functioning of epidermal growth factor receptors was required, or even involved, in masking.

The circadian Clock mutant mouse: impaired masking response to light.

Synchronization of an internal clock (entrainment) and a direct response to light (masking) are complementary ways of restricting activity of an animal to day or night. The protein CLOCK has an important role in the oscillatory mechanism of mammalian pacemakers. Our data show that it is also involved in masking responses. Mice with the Clock/Clock mutation reduced their wheel running less than wildtypes when given 1-h light pulses of light (2-1,600 lx) in the night. With dimmer lights (<2 lx), there were no significant differences between mutant and wildtype mice. Impaired masking responses to light in Clock/Clock mice were confirmed in tests with ultradian light-dark cycles (3.5:3.5 h and 1:1 h). Tests with pulses of light longer than 1 h revealed that, although the mutants responded more slowly to light, they sustained the suppression of activity over the course of the 3-h tests better than wildtypes.

Nocturnal activity in a diurnal rodent (Arvicanthis niloticus): the importance of masking.

It is known that day-active Nile grass rats, Arvicanthis niloticus, increase the amount of activity in the night relative to that in the day when provided with running wheels. This was confirmed in the present study. Animals without a wheel displayed 69.0% of their general activity in the L phase of a 12:12 h light-dark cycle; animals provided with wheels had only 48.6% of their wheel revolutions in the light. The contribution of direct (masking) responses to light to the increased nocturnality of animals with wheels was examined in two experiments. In experiment 1, masking was tested by exposing the animals to repeated cycles of 30 min of entraining light and 30 min of a different, usually dimmer light, during the L phase of a 12:12 h light-dark cycle. For animals with wheels, there was more running during the 30-min pulses of dim light or darkness than during the 30-min periods of entraining light. In contrast, for animals without wheels, there was more general activity during the 30-min periods of entraining light than during the 30-min pulses of dim light or darkness. In experiment 2, the animals were first exposed to a 12:12 h light-dark cycle and then put on a 1:10:1:12 h LDLD skeleton photoperiod. Animals with wheels increased their running during the subjective day of the skeleton photoperiod compared to that in the actual day of the 12:12 h light-dark cycle. Animals without wheels showed similar levels of general activity during the subjective day of the skeleton photoperiod and the actual day of the 12:12 h cycle. These experiments demonstrate that when Nile rats have running wheels, their increased nocturnal activity is associated with an increased suppression of locomotion in direct response to light. It is possible that changes in masking responses to light may be an essential and integral component of switching between diurnal and nocturnal activity profiles.

Impaired masking responses to light in melanopsin-knockout mice.

There are two ways in which an animal can confine its behavior to a nocturnal or diurnal niche. One is to synchronize an endogenous clock that in turn controls the sleep-wake cycle. The other is to respond directly to illumination with changes in activity. In mice, high illumination levels suppress locomotion (negative masking) and low illumination levels enhance locomotion (positive masking). To investigate the role of the newly discovered opsin-like protein melanopsin in masking, we used 1 h and 3 h pulses of light given in the night, and also a 3.5:3.5 h light-dark (LD) cycle. Mice lacking the melanopsin gene had normal enhancement of locomotion in the presence of dim lights but an impaired suppression of locomotion in the presence of bright light. This impairment was evident only with lights in the order of 10 lux or brighter. This suggests that melanopsin in retinal ganglion cells is involved in masking, as it is in pupil contraction and phase shifts. Melanopsin is especially important in maintaining masking responses over long periods.

Period gene expression in the suprachiasmatic nucleus of behaviorally decoupled hamsters.

Circadian rhythms in locomotor activity in nocturnal animals typically show activity during the night followed by quiescence during the day. In hamsters, this activity pattern can be altered by confining them to novel running wheels for a few hours during the day, thereby inducing them to be active. After several days of induced activity, two bouts of activity occur spontaneously, one during the day, and the other at night. This phenomenon is known as behavioral decoupling. In the present study we examined the effects of behavioral decoupling on the pattern of SCN Per expression. Our results show robust expression of Period genes in the SCN of animals killed during either of the two inactive phases within a 24-h period, and weak expression of those genes in the SCN of animals killed during the two active phases. This contrasts with the circadian rhythm of Per expression typically seen in the SCN of entrained or free running animals. Our data support the idea that behavioral decoupling reorganizes the circadian system in a manner different from that produced by constant light-induced splitting and constant dark splitting.

Melanopsin and rod-cone photoreceptive systems account for all major accessory visual functions in mice.

In the mammalian retina, besides the conventional rod-cone system, a melanopsin-associated photoreceptive system exists that conveys photic information for accessory visual functions such as pupillary light reflex and circadian photo-entrainment. On ablation of the melanopsin gene, retinal ganglion cells that normally express melanopsin are no longer intrinsically photosensitive. Furthermore, pupil reflex, light-induced phase delays of the circadian clock and period lengthening of the circadian rhythm in constant light are all partially impaired. Here, we investigated whether additional photoreceptive systems participate in these responses. Using mice lacking rods and cones, we measured the action spectrum for phase-shifting the circadian rhythm of locomotor behaviour. This spectrum matches that for the pupillary light reflex in mice of the same genotype, and that for the intrinsic photosensitivity of the melanopsin-expressing retinal ganglion cells. We have also generated mice lacking melanopsin coupled with disabled rod and cone phototransduction mechanisms. These animals have an intact retina but fail to show any significant pupil reflex, to entrain to light/dark cycles, and to show any masking response to light. Thus, the rod-cone and melanopsin systems together seem to provide all of the photic input for these accessory visual functions.

Behavioral arousal blocks light-induced phase advances in locomotor rhythmicity but not light-induced Per1 and Fos expression in the hamster suprachiasmatic nucleus.

Both photic and nonphotic stimuli entrain circadian rhythms. Although the adaptive significance of nonphotic clock resetting is unknown, one possibility is that nonphotic cues modulate circadian responses to light. Results of studies on the interaction between photic and nonphotic stimuli support this idea. During the day, light blocks the effects of nonphotic stimuli on the phase of locomotor rhythms and on expression of clock genes in suprachiasmatic nucleus (SCN) neurons. At night, novelty-induced activity prior to and during exposure to light attenuates the phase-shifting response to that light, but the effects of this manipulation on clock gene expression are unknown. The present experiments explore the interaction between behavioral state and response to light at the molecular level. We show that confining hamsters to novel wheels immediately after a light pulse during the late subjective night attenuates light-induced phase advances of wheel-running rhythms and the transient effects on circadian period. In contrast to the striking effect of novelty-induced activity on behavioral responses to light, Fos protein and Per1 mRNA were robustly expressed in the SCN of all light-pulsed animals, regardless of behavioral treatment. Our results are inconsistent with the idea that light and nonphotic stimuli block each other's effects on phase shifts by inducing or attenuating transcription of Per1. Photic regulation of clock genes and spontaneous rhythmic expression of clock genes are probably mediated by different mechanisms.

Beyond the suprachiasmatic nucleus.

The suprachiasmatic nucleus is the master oscillator controlling circadian rhythms in mammals. Yet extensive temporal restructuring of behavior can occur without participation of the suprachiasmatic nucleus. This raises questions about current thinking about how to cope with jet lag and shift work.