Effects of Fruit Toxins on Intestinal and Microbial ß-Glucosidase Activities of Seed-Predating and Seed-Dispersing Rodents (Acomys spp.).

Plant secondary compounds (PSCs) have profound influence on the ecological interaction between plants and their consumers. Glycosides, a class of PSC, are inert in their intact form and become toxic on activation by either plant ß-glucosidase enzymes or endogenous ß-glucosidases produced by the intestine of the plant-predator or its microbiota. Many insect herbivores decrease activities of endogenous ß-glucosidases to limit toxin exposure. However, such an adaptation has never been investigated in nonmodel mammals. We studied three species of spiny mice (Acomys spp.) that vary in their feeding behavior of the glycoside-rich fruit of Ochradenus baccatus. Two species, the common (Acomys cahirinus) and Crete (Acomys minous) spiny mice, behaviorally avoid activating glycosides, while the golden spiny mouse (Acomys russatus) regularly consumes activated glycosides. We fed each species a nontoxic diet of inert glycosides or a toxic diet of activated fruit toxins and investigated the responses of intestinal and microbial ß-glucosidase activities. We found that individuals feeding on activated toxins had lower intestinal ß-glucosidase activity and that the species that behaviorally avoid activating glycosides also had lower intestinal ß-glucosidase activity regardless of treatment. The microbiota represented a larger source of toxin liberation, and the toxin-adapted species (golden spiny mouse) exhibited almost a fivefold increase in microbial ß-glucosidase when fed activated toxins, while other species showed slight decreases. These results are contrary to those in insects, where glycoside-adapted species have lower ß-glucosidase activity. The glycoside-adapted golden spiny mouse may have evolved tolerance mechanisms such as enhanced detoxification rather than avoidance mechanisms.

Geographic variation in thermal tolerance and strategies of heat shock protein expression in the land snail Theba pisana in relation to genetic structure.

Land snails are exposed to conditions of high ambient temperature and low humidity, and their survival depends on a suite of morphological, behavioral, physiological, and molecular adaptations to the specific microhabitat. We tested in six populations of the land snail Theba pisana whether adaptations to different habitats affect their ability to cope with thermal stress and their strategies of heat shock protein (HSP) expression. Levels of Hsp70 and Hsp90 in the foot tissue were measured in field-collected snails and after acclimation to laboratory conditions. Snails were also exposed to various temperatures (32 up to 54 °C) for 2 h and HSP messenger RNA (mRNA) levels were measured in the foot tissue and survival was determined. To test whether the physiological and molecular data are related to genetic parameters, we analyzed T. pisana populations using partial sequences of nuclear and mitochondrial DNA ribosomal RNA genes. We show that populations collected from warmer habitats were more thermotolerant and had higher constitutive levels of Hsp70 isoforms in the foot tissue. Quantitative real-time polymerase chain reaction (PCR) analysis indicated that hsp70 and hsp90 mRNA levels increased significantly in response to thermal stress, although the increase in hsp70 mRNA was larger compared to hsp90 and its induction continued up to higher temperatures. Generally, warm-adapted populations had higher temperatures of maximal induction of hsp70 mRNA synthesis and higher upper thermal limits to HSP mRNA synthesis. Our study suggests that Hsp70 in the foot tissue of T. pisana snails may have important roles in determining stress resistance, while Hsp90 is more likely implicated in signal transduction processes that are activated by stress. In the phylogenetic analysis, T. pisana haplotypes were principally divided into two major clades largely corresponding to the physiological ability to withstand stress, thus pointing to genetically fixed tolerance.

Small intestinal hydrolysis of plant glucosides: higher glucohydrolase activities in rodents than passerine birds.

Glycosides are a major group of plant secondary compounds characterized by one or more sugars conjugated to a lipophilic, possibly toxic aglycone, which is released upon hydrolysis. We compared small intestinal homogenate hydrolysis activity of three rodent and two avian species against four substrates: amygdalin and sinigrin, two plant-derived glucosides, the sugar lactose, whose hydrolysis models some activity against flavonoid and isoflavonoid glucosides, and the disaccharide sugar maltose (from starch), used as a comparator. Three new findings extend our understanding of physiological processing of plant glucosides: (1) the capacity of passerine birds to hydrolyze plant glucosides seems relatively low, compared with rodents; (2) in this first test of vertebrates' enzymic capacity to hydrolyze glucosinolates, sinigrin hydrolytic capacity seems low; (3) in laboratory mice, hydrolytic activity against lactose resides on the enterocytes' apical membrane facing the intestinal lumen, but activity against amygdalin seems to reside inside enterocytes.

Natural variation in resistance to desiccation and heat shock protein expression in the land snail Theba pisana along a climatic gradient.

Land snails frequently encounter desiccating conditions, and their survival depends on a suite of morphological, physiological, and molecular adaptations to the specific microhabitat. Strategies of survival can be determined by integrating information from various levels of biological organization. In this study, we used a combination of physiological parameters related to water economy and molecular factors (stress protein expression) to investigate the strategies of survival adopted by seven populations of the Mediterranean-type land snail Theba pisana from different habitats. We analyzed water compartmentalization during aestivation and used experimental desiccation to compare desiccation resistance. We also measured the endogenous levels of heat shock proteins (HSPs) Hsp72, Hsp74, and Hsp90 under nonstress conditions and analyzed the HSP response to desiccation in two populations that differed mostly in their resistance to desiccation. We revealed significant intraspecific differences in resistance to desiccation that seem to be determined by the speed of recruitment of the water-preserving mechanisms. The ability to cope with desiccating conditions was correlated with habitat temperature but not with the rainfall gradient, implying that in the coastal region, temperature is likely to have a major impact on desiccation resistance rather than precipitation. Also, higher desiccation resistance was correlated with higher constitutive levels of Hsp74 in the foot tissue. HSPs were upregulated during desiccation, but the response was delayed and was milder in the most resistant population compared to the most susceptible one. Our study suggests that T. pisana populations from warmer habitats were more resistant to desiccation and developed distinct strategies of HSP expression for survival, namely, the maintenance of high constitutive levels of Hsp70 together with a delayed and limited response to stress.

Taste and physiological responses to glucosinolates: seed predator versus seed disperser.

In contrast to most other plant tissues, fleshy fruits are meant to be eaten in order to facilitate seed dispersal. Although fleshy fruits attract consumers, they may also contain toxic secondary metabolites. However, studies that link the effect of fruit toxins with seed dispersal and predation are scarce. Glucosinolates (GLSs) are a family of bitter-tasting compounds. The fleshy fruit pulp of Ochradenus baccatus was previously found to harbor high concentrations of GLSs, whereas the myrosinase enzyme, which breaks down GLSs to produce foul tasting chemicals, was found only in the seeds. Here we show the differential behavioral and physiological responses of three rodent species to high dose (80%) Ochradenus' fruits diets. Acomys russatus, a predator of Ochradenus' seeds, was the least sensitive to the taste of the fruit and the only rodent to exhibit taste-related physiological adaptations to deal with the fruits' toxins. In contrast, Acomys cahirinus, an Ochradenus seed disperser, was more sensitive to a diet containing the hydrolyzed products of the GLSs. A third rodent (Mus musculus) was deterred from Ochradenus fruits consumption by the GLSs and their hydrolyzed products. We were able to alter M. musculus avoidance of whole fruit consumption by soaking Ochradenus fruits in a water solution containing 1% adenosine monophosphate, which blocks the bitter taste receptor in mice. The observed differential responses of these three rodent species may be due to evolutionary pressures that have enhanced or reduced their sensitivity to the taste of GLSs.

Physiological and behavioural effects of fruit toxins on seed-predating versus seed-dispersing congeneric rodents.

Fleshy, ripe fruits attract seed dispersers but also seed predators. Although many fruit consumers (legitimate seed dispersers as well as seed predators) are clearly exposed to plant secondary compounds (PSCs), their impact on the consumers' physiology and foraging behaviour has been largely overlooked. Here, we document the divergent behavioural and physiological responses to fruit consumption of three congeneric rodent species in the Middle East, representing both seed dispersers and seed predators. The fruit pulp of the desert plant Ochradenus baccatus contains high concentrations of glucosinolates (GLSs). These GLSs are hydrolyzed into active toxic compounds upon contact with the myrosinase enzyme released from seeds crushed during fruit consumption. Acomys russatus and A. cahirinus share a desert habitat. Acomys russatus acts as an O. baccatus seed predator, and A. cahirinus circumvents the activation of the GLSs by orally expelling vital seeds. We found that between the three species examined, A. russatus was physiologically most tolerant to whole fruit consumption and even A. minous, which is evolutionarily naïve to O. baccatus, exhibits greater tolerance to whole fruit consumption than A. cahirinus. However, like A. cahirinus, A. minous may also behaviourally avoid the activation of the GLSs by making a hole in the pulp and consuming only the seeds. Our findings demonstrate that seed predators have a higher physiological tolerance than seed dispersers when consuming fruits containing toxic PSCs. The findings also demonstrate the extreme ecological/evolutionary lability of this plant-animal symbiosis to shift from predation to mutualism and vice versa.

Intraspecific directed deterrence by the mustard oil bomb in a desert plant.

Plant secondary metabolites (SMs) acting as defensive chemicals in reproductive organs such as fruit tissues play roles in both mutualistic and antagonistic interactions between plants and seed dispersers/predators. The directed-deterrence hypothesis states that SMs in ripe fruits deter seed predators but have little or no effect on seed dispersers. Indeed, studies have demonstrated that birds are able to cope with fruit SMs whereas rodents are deterred by them. However, this mechanism was only demonstrated at the class level, i.e., between birds and mammals, based on differences in the vanilloid receptors. Here we present experimental and behavioral data demonstrating the use of the broad-range, class-independent "mustard oil bomb" mechanism in Ochradenus baccatus fruits to force a behavioral change at an ecological timescale, converting rodents from seed predators to seed dispersers. This is achieved by a unique compartmentalization of the mustard oil bomb, causing activation of the system only upon seed and pulp coconsumption, encouraging seed dispersal via seed spitting by rodents. Our findings demonstrate the power of SMs to shift the animal-plant relationship from predation to mutualism and provide support for the directed-deterrence hypothesis at the intraspecific level, in addition to the interspecific level.

The heat shock response in congeneric land snails (Sphincterochila) from different habitats.

Land snails are subject to daily and seasonal variations in temperature and in water availability, and use heat shock proteins (HSPs) as part of their survival strategy. We used experimental heat stress to test whether adaptation to different habitats affects HSP expression in two closely related Sphincterochila snail species, a desert species, Sphincterochila zonata, and a Mediterranean-type species, Sphincterochila cariosa. Our findings show that in S. cariosa, heat stress caused rapid induction of Hsp70 proteins and Hsp90 in the foot and kidney tissues, whereas the desert-inhabiting species S. zonata displayed delayed induction of Hsp70 proteins in the foot and upregulation of Hsp90 alone in the kidney. Our study suggests that Sphincterochila species use HSPs as part of their survival strategy following heat stress and that adaptation to different habitats results in the development of distinct strategies of HSP expression in response to heat, namely the reduced induction of HSPs in the desert-dwelling species. We suggest that the desert species S. zonata relies on mechanisms and adaptations other than HSP induction, thus avoiding the fitness consequences of continuous HSP upregulation.

Heat shock proteins and survival strategies in congeneric land snails (Sphincterochila) from different habitats.

Polmunate land snails are subject to stress conditions in their terrestrial habitat, and depend on a range of behavioural, physiological and biochemical adaptations for coping with problems of maintaining water, ionic and thermal balance. The involvement of the heat shock protein (HSP) machinery in land snails was demonstrated following short-term experimental aestivation and heat stress, suggesting that land snails use HSPs as part of their survival strategy. As climatic variation was found to be associated with HSP expression, we tested whether adaptation of land snails to different habitats affects HSP expression in two closely related Sphincterochila snail species, a desert species Sphincterochila zonata and a Mediterranean-type species Sphincterochila cariosa. Our study suggests that Sphincterochila species use HSPs as part of their survival strategy following desiccation and heat stress, and as part of the natural annual cycle of activity and aestivation. Our studies also indicate that adaptation to different habitats results in the development of distinct strategies of HSP expression in response to stress, namely the reduced expression of HSPs in the desert-inhabiting species. We suggest that these different strategies reflect the difference in heat and aridity encountered in the natural habitats, and that the desert species S. zonata relies on mechanisms and adaptations other than HSP induction thus avoiding the fitness consequences of continuous HSP upregulation.

Capacity for absorption of water-soluble secondary metabolites greater in birds than in rodents.

Plant secondary metabolites (SMs) are pervasive in animal foods and potentially influence feeding behavior, interspecies interactions, and the distribution and abundance of animals. Some of the major classes of naturally occurring SMs in plants include many water-soluble compounds in the molecular size range that could cross the intestinal epithelium via the paracellular space by diffusion or solvent drag. There are differences among species in paracellular permeability. Using Middle Eastern rodent and avian consumers of fruits containing SMs, we tested the hypothesis that avian species would have significantly higher paracellular permeability than rodent species. Permeability in intact animals was assessed using standard pharmacological methodology to measure absorption of two radiolabeled, inert, neutral water-soluble probes that do not interact with intestinal nutrient transporters, L-arabinose (M(r) = 150.1 Da) and lactulose (M(r) = 342.3 Da). We also measured absorption of labeled 3-O-methyl-D-glucose (3OMD-glucose; M(r) = 194.2 Da), which is a nonmetabolized analogue of D-glucose that is passively absorbed through the paracellular space but also transported across the enterocyte membranes. Most glucose was absorbed by all species, but arabinose fractional absorption (f) was nearly three times higher in birds (1.03±0.17, n = 15 in two species) compared to rodents (0.37±0.06, n = 10 in two species) (P<0.001). Surprisingly, the apparent rates of absorption in birds of arabinose exceeded those of 3OMD-glucose. Our findings are in agreement with previous work showing that the paracellular pathway is more prominent in birds relative to nonflying mammals, and suggests that birds may be challenged by greater absorption of water-soluble, dietary SMs. The increased expression of the paracellular pathway in birds hints at a tradeoff: the free energy birds gain by absorbing water-soluble nutrients passively may be offset by the metabolic demands placed on them to eliminate concomitantly absorbed SMs.

Heat shock protein expression in relation to reproductive cycle in land snails: Implications for survival.

Land snails are subject to daily and seasonal variations in temperature and in water availability and use heat shock proteins (HSPs) as part of their survival strategy. We tested whether the reproductive cycle of land snails affects the endogenous levels of HSPs, and their involvement in the reproductive process. We examined HSP levels in the foot tissue of two Sphincterochila species, S. cariosa and S. zonata, before and after laying eggs, and analyzed the albumen gland (reproductive organ) of both species and eggs of S. cariosa for the presence and quantity of various HSPs. Our study shows reduction in the expression level of Hsp70 isoforms and Hsp90 in S. zonata foot and of Hsp74 in S. cariosa foot during the period preceding egg laying compared to the post-reproductive stage. Hsp70 isoforms and Hsp25 were highly expressed in both large albumen glands and in freshly laid eggs of S. cariosa, whereas large albumen glands of S. zonata expressed mainly Hsp70 isoforms. We conclude that a trade-off between survival and fertility is responsible for the expression level of HSPs in the foot tissue of Sphincterochila snails. Our study shows that HSPs are involved in the reproductive process. We propose that parental provision of HSPs may be part of a "be prepared" strategy of Sphincterochila snails, and that HSPs may play important roles in the survival strategy of land snails during the early life stages. Our observations also highlight the importance of the reproductive status in study of whole organisms, especially when assessing the HSP response to stress.

Natural annual cycle of heat shock protein expression in land snails: desert versus Mediterranean species of Sphincterochila.

Land snails are subject to daily and seasonal variations in temperature and in water availability, and have evolved annual cycles of activity and aestivation as part of their survival strategy. We tested in the field whether adaptation to different habitats affects the endogenous levels of heat shock proteins (HSPs) in two closely related Sphincterochila snail species, a desiccation-resistant desert species, Sphincterochila zonata, and a Mediterranean-type, desiccation-sensitive species, S. cariosa. We examined HSP levels in various tissues of snails during aestivation and after resumption of activity. Our study shows that, during aestivation, S. cariosa had higher standing stocks of Hsp70 in the foot and the hepatopancreas, and of small HSPs (sHSPs) in all the examined tissues, whereas S. zonata had higher stocks of Hsp70 in the kidney and of Hsp90 in the kidney and in the hepatopancreas. Arousal induced a general upregulation of HSPs, except for Hsp90, the expression of which in the foot was higher during aestivation. We suggest that the stress protein machinery is upregulated during arousal in anticipation of possible oxidative stress ensuing from the accelerating metabolic rate and the exit from the deep hypometabolic state. Our findings support the concept that, in land snails, aestivation and activity represent two distinct physiological states, and suggest that land snails use HSPs as important components of the aestivation mechanism, and as part of their survival strategy during and after arousal. Our study also indicates that adaptation to different habitats results in the development of distinct strategies of HSP expression with likely consequences for the ecology and distribution of land snails.

Heat shock proteins and resistance to desiccation in congeneric land snails.

Land snails are subject to daily and seasonal variations in temperature and in water availability and depend on a range of behavioral and physiological adaptations for coping with problems of maintaining water, ionic, and thermal balance. Heat shock proteins (HSPs) are a multigene family of proteins whose expression is induced by a variety of stress agents. We used experimental desiccation to test whether adaptation to different habitats affects HSP expression in two closely related Sphincterochila snail species, a desiccation-resistant, desert species Sphincterochila zonata, and a Mediterranean-type, desiccation-sensitive species Sphincterochila cariosa. We examined the HSP response in the foot, hepatopancreas, and kidney tissues of snails exposed to normothermic desiccation. Our findings show variations in the HSP response in both timing and magnitude between the two species. The levels of endogenous Hsp72 in S. cariosa were higher in all the examined tissues, and the induction of Hsp72, Hsp74, and Hsp90 developed earlier than in S. zonata. In contrary, the induction of sHSPs (Hsp25 and Hsp30) was more pronounced in S. zonata compared to S. cariosa. Our results suggest that land snails use HSPs as part of their survival strategy during desiccation and as important components of the aestivation mechanism in the transition from activity to dormancy. Our study underscores the distinct strategy of HSP expression in response to desiccation, namely the delayed induction of Hsp70 and Hsp90 together with enhanced induction of sHSPs in the desert-dwelling species, and suggests that evolution in harsh environments will result in selection for reduced Hsp70 expression.

Consequences of buoyancy to the maneuvering capabilities of a foot-propelled aquatic predator, the great cormorant (Phalcrocorax carbo sinensis).

Great cormorants are foot-propelled aquatic divers utilizing a region of the water column where their underwater foraging behavior is affected by their buoyancy. While swimming horizontally underwater, cormorants use downward lift forces generated by their body and tail to overcome their buoyancy. Here we assess the potential of this swimming strategy for controlling maneuvers in the vertical plane. We recorded the birds swimming through a submerged obstacle course and analyzed their maneuvers. The birds reduced swimming speed by only 12% to maneuver and were able to turn upward and then downward in the sagittal plane at a minimal turning radius of 32+/-4 cm (40% body length). Using a quasi-steady approach, we estimated the time-line for hydrodynamic forces and the force-moments produced while maneuvering. We found that the tail is responsible for the pitch of the body while motions of the body, tail, neck and feet generate forces normal (vertically) to the swimming direction that interact with buoyancy to change the birds' trajectory. Vertical maneuvers in cormorants are asymmetric in energy cost. When turning upward, the birds use their buoyancy but they must work harder to turn downward. Lift forces generated by the body were always directed ventrally. Propulsion improves the ability to make tight turns when the center of the turn is ventral to the birds. The neck produced only a small portion (10%) of the normal vertical forces but its length may allow prey capture at the end of pursuit, within the minimum turning radius.

Prey detection by great cormorant (Phalacrocorax carbo sinensis) in clear and in turbid water.

The scattering and absorption of light by water molecules and by suspended and dissolved matter (turbidity) degrade image transmission and, thus, underwater perception. We tested the effects on visual detection of prey size and distance (affecting apparent prey size) and of low-level water turbidity in hand-reared great cormorants (Phalacrocorax carbo sinensis) diving for natural prey (fish) in a forced-choice situation. The cormorants' detection of underwater prey relied on vision. The minimal tested subtending visual angle of the prey at detection ranged between approximately 34.2' (prey size constant; distance varied) and 9.5' (distance constant; prey size varied). For all tested distances (0.8-3.1 m) the mean detection success was significantly higher than the chance level. The probability of a correct choice declined significantly with increased distance, with Detection success=-0.034D+1.021 (where D is distance, r(2)=0.5, N=70, P<0.001). The combined effect of turbidity and distance on the probability of detection success was significant, with both variables having a negative effect: Detection success=-0.286D-0.224Tu+1.691 (where Tu is turbidity, r(2)=0.68, N=144, P<0.001). At prey detection threshold, the relationship between distance and turbidity was: D=3.79e(-4.55Tu). It is concluded that (i) the subtending angle of natural prey at detection was lower than that of resolution of square-wave, high-contrast grating and (ii) turbidity, at levels significantly lower than commonly used in behavioural experiments, had a pronounced effect on visually mediated behaviour patterns.

Dietary protein influences the rate of 15N incorporation in blood cells and plasma of Yellow-vented bulbuls (Pycnonotus xanthopygos).

The rate at which an animal's tissues incorporate the isotopic composition of food determines the time window during which ecologists can discern diet changes. We investigated the effect of protein content in the diet on the incorporation rate of (15)N into the plasma proteins and blood cells of Yellow-vented bulbuls (Pycnonotus xanthopygos). Using model comparison analyses, we found that one-compartment models described incorporation data better than two-compartment models. Dietary protein content had a significant effect on the residence time of (15)N in plasma proteins and blood cells. The diet with the highest protein content led to a (15)N retention time of 21 and 5 days for cells and plasma, respectively. In contrast, average (15)N retention time in the cells and plasma of birds fed on the diet with the lowest protein was 31 and 7 days, respectively. The isotopic discrimination factor Delta(15)N=delta(15)N(tissues)-delta(15)N(diet) was also dependent on dietary protein content, and was lowest in birds fed the diet with the highest protein content. Blood, plasma and excreta were enriched in (15)N relative to diet. In contrast, ureteral urine was either significantly depleted of (15)N in birds fed the diet with the lowest protein content or did not differ in delta(15)N from the diets with the intermediate and high protein content. Thus, isotopic incorporation rates and tissue-to-diet discrimination factors cannot be considered fixed, as they depend on diet composition.

The relationship between uric acid and its oxidative product allantoin: a potential indicator for the evaluation of oxidative stress in birds.

Uric acid is the main nitrogenous waste product in birds but it is also known to be a potent antioxidant. Hominoid primates and birds lack the enzyme urate oxidase, which oxidizes uric acid to allantoin. Consequently, the presence of allantoin in their plasma results from non-enzymatic oxidation. In humans, the allantoin to uric acid ratio in plasma increases during oxidative stress, thus this ratio has been suggested to be an in vivo marker for oxidative stress in humans. We measured the concentrations of uric acid and allantoin in the plasma and ureteral urine of white-crowned sparrows (Zonotrichia leucophrys gambelii) at rest, immediately after 30 min of exercise in a hop/hover wheel, and after 1 h of recovery. The plasma allantoin concentration and the allantoin to uric acid ratio did not increase during exercise but we found a positive relationship between the concentrations of uric acid and allantoin in the plasma and in the ureteral urine in the three activity phases. In the plasma, the slope of the regression describing the above positive relationships was significantly higher immediately after activity. We suggest that the slope indicates the rate of uric acid oxidation and that during activity this rate increases as a result of higher production of free radicals. The present study demonstrates that allantoin is present in the plasma and in the ureteral urine of white-crowned sparrows and therefore might be useful as an indicator of oxidative stress in birds.

Reciprocal activation of HSF1 and HSF3 in brain and blood tissues: is redundancy developmentally related?

Transcriptional induction of heat-shock genes in response to temperature elevation and other stresses is mediated by heat-shock transcription factors (HSFs). Avian cells express two redundant heat-shock responsive factors, HSF1 and HSF3, which differ in their activation kinetics and threshold induction temperature. Unlike the ubiquitous activation of HSF1, the DNA-binding activity of HSF3 is restricted to undifferentiated avian cells and embryonic tissues. Herein, we report a reciprocal activation of HSF1 and HSF3 in vivo. Whereas HSF1 mediates transcriptional activity only in the brain upon severe heat shock, HSF3 is exclusively activated in blood cells upon light, moderate, and severe heat shock, promoting induction of heat-shock genes. Although not activated, HSF1 is expressed in blood cell nuclei in a granular appearance, suggesting regulation of genes other than heat-shock genes. Intraspecific comparison of heat-sensitive and heat-resistant fowl strains indicates that the unique activation pattern of HSF3 in blood tissue is a general phenomenon, not related to thermal history. Taken together, HSF1 and HSF3 mediate transcriptional activity of adult tissues and differentiated cells in a nonredundant manner. Instead, an exclusive, tissue-specific activation is observed, implying that redundancy may be developmentally related. The physiological and developmental implications are discussed.

Submerged swimming of the great cormorant Phalacrocorax carbo sinensis is a variant of the burst-and-glide gait.

Cormorants are water birds that forage by submerged swimming in search and pursuit of fish. Underwater they swim by paddling with both feet simultaneously in a gait that includes long glides between consecutive strokes. At shallow swimming depths the birds are highly buoyant as a consequence of their aerial lifestyle. To counter this buoyancy cormorants swim underwater with their body at an angle to the swimming direction. This mechanical solution for foraging at shallow depth is expected to increase the cost of swimming by increasing the drag of the birds. We used kinematic analysis of video sequences of cormorants swimming underwater at shallow depth in a controlled research setup to analyze the swimming gait and estimate the resultant drag of the birds during the entire paddling cycle. The gliding drag of the birds was estimated from swimming speed deceleration during the glide stage while the drag during active paddling was estimated using a mathematical ;burst-and-glide' model. The model was originally developed to estimate the energetic saving from combining glides with burst swimming and we used this fact to test whether the paddling gait of cormorants has similar advantages. We found that swimming speed was correlated with paddling frequency (r=0.56, P<0.001, N=95) where the increase in paddling frequency was achieved mainly by shortening the glide stage (r=-0.86, P<0.001, N=95). The drag coefficient of the birds during paddling was higher on average by two- to threefold than during gliding. However, the magnitude of the drag coefficient during the glide was positively correlated with the tilt of the body (r=0.5, P<0.003, N=35) and negatively correlated with swimming speed (r=-0.65, P<0.001, N=35), while the drag coefficient during the stroke was not correlated with tilt of the body (r=-0.11, P>0.5, N=35) and was positively correlated with swimming speed (r=0.41, P<0.015, N=35). Therefore, the difference between the drag coefficient during the glide and during propulsion diminished at lower speeds and larger tilt. The mean drag of the birds for a single paddling cycle at an average swimming speed of 1.5 m s(-1) was 5.5+/-0.68 N. The burst-and-glide model predicts that energy saving from using burst-and-glide in the paddling cycle is limited to relatively fast swimming speeds (>1.5 m s(-1)), but that as the birds dive deeper (>1 m where buoyancy is reduced), the burst-and-glide gait may become beneficial even at lower speeds.

Ectothermy and endothermy: evolutionary perspectives of thermoprotection by HSPs.

Living organisms respond to heat exposure by selectively expressing heat shock proteins (HSPs). Accumulation of HSPs confers thermotolerance in cell cultures and in ectotherms and is an important component of the heat shock response. This response, however, has not been directly examined in relation to different ;thermal states', namely ectothermy vs endothermy. By using avian development as a model system for transition from ectothermy to endothermy, we show that, in contrast to the ectothermic state, in the endothermic state the organism is more resistant to heat but relies less on HSPs as a first-line thermoprotective mechanism. Moreover, intraspecific, real-time, in vivo measurements in genetically diverse fowl strains relate improvement of thermoresistance in endotherms to improved body temperature (Tb) regulation, with a concomitant delay in the expression of HSPs. The time course of this delay and the Tb at which it occurs imply that the ontogenetic and evolutionary pathways leading to improved thermoresistance may have followed two, apparently non-related, parallel routes--cellular and peripheral (non-cellular). In search of other cellular components that differentially participate in the heat shock response, we revealed a significant expression of fatty acid synthase (FAS) in heat-exposed endotherms but not in ectotherms.