Context dependency and generality of fever in insects.

Fever can reduce mortality in infected animals. Yet, despite its fitness-enhancing qualities, fever often varies among animals. We used several approaches to examine this variation in insects. Texas field crickets (Gryllus texensis) exhibited a modest fever (1 °C increase in preferred body temperature, T pref) after injection of prostaglandin, which putatively mediates fever in both vertebrates and invertebrates, but they did not exhibit fever during chronic exposure to heat-killed bacteria. Further, chronic food limitation and mating status did not affect T pref or the expression of behavioural fever, suggesting limited context dependency of fever in G. texensis. Our meta-analysis of behavioural fever studies indicated that behavioural fever occurs in many insects, but it is not ubiquitous. Thus, both empirical and meta-analytical results suggest that the fever response in insects 'is widespread, although certainly not inevitable' (Moore 2002). We highlight the need for future work focusing on standardizing an experimental protocol to measure behavioural fever, understanding the specific mechanism(s) underlying fever in insects, and examining whether ecological or physiological costs often outweigh the benefits of fever and can explain the sporadic nature of fever in insects.

Why should an immune response activate the stress response? Insights from the insects (the cricket Gryllus texensis).

Mediators of the stress response (e.g. glucocorticoids and norepinephrine) can be immunosuppressive. Nevertheless, immune challenge leads to the release of these compounds in vertebrates. To resolve this paradox, it has been suggested that stress hormones help restore immune homeostasis, preventing self-damage. A comparative approach may provide additional hypotheses as to why an immune challenge induces the release of stress hormones/neurohormones. Octopamine, a neurohormonal mediator of the stress response in the cricket Gryllus texensis, increased in concentration in the hemolymph during an immune challenge. Therefore, the release of stress hormones during an immune response occurs in animals across phyla. Octopamine induced an increase in lipid concentration in the hemolymph. After an acute stress (flying or running) the total number of hemocytes in the hemolymph increased. Injections of octopamine had the same effect, suggesting that it may enhance hemocyte-dependent immune functions. On the other hand, octopamine decreased lysozyme-like activity in vitro, suggesting that it inhibits some immune functions. However, lysozyme-like activity was increased by the presence of heat-killed bacteria in vitro and this increase was significantly augmented by the presence of octopamine. Therefore, the effect of octopamine on immune function differed depending on the presence of pathogens. Stress hormones may help shift immune function into the most optimal configuration depending on the physiological context.

Immune lysis of lipid vesicles containing myelin basic protein or glycolipid antigens by multiple sclerosis and normal sera.

We have compared the reactivity of sera from 34 multiple sclerosis (MS) patients and 32 normal (N) individuals with lipid vesicles containing myelin basic protein (BP) and several glycolipids reconstituted into a membrane environment. The ability of the sera to cause complement-mediated lysis of lipid vesicles containing these antigens was determined by measuring the release of a water-soluble spin label, tempocholine chloride, from the height of its electron spin resonance spectrum. Only 4 MS sera caused lysis of BP-containing vesicles which was comparable to that produced by specific antibody to BP. A number of both MS and N sera caused significant lysis of vesicles containing GM1 ganglioside or digalactosyldiglyceride. A few MS and N sera also caused significant lysis of vesicles containing GM2, GT1 and GD1a gangliosides. However, in no case was there a statistically significant difference between the mean lysis produced by MS and N sera. There was some overlap between the specific MS and N sera reactive to vesicles containing BP, GM1, GM2, and DGDG while a completely different group of MS and N sera were reactive to GT1 and GD1a gangliosides. This suggested that there was either antigenic cross reactivity between the two groups of glycolipids or two different origins of the immune response to the two groups of antigens. It was concluded that antibody-dependent complement fixation by these particular antigens, in the kind of lipid environment used, is not characteristic of or specific to MS.

The specificity of behavioral fever in the cricket Acheta domesticus.

When infected, some insects can raise their body temperature by moving to warmer areas. This behavioral fever response can help the host overcome infection. However, not all parasites and pathogens are equally susceptible to increases in host temperature. Elevating the temperature of the cricket Acheta domesticus from room temperature (22 C) to 33 C did not reduce the survival of parasitoid flies or reduce the number of gregarine gut protozoans, and crickets infested with these parasites showed no increase in their temperature preference. Warmer temperatures (33 C) did not increase the survival of crickets infected with the bacterium Serratia marcescens, and infected crickets did not prefer warmer temperatures. However crickets infected with the intracellular parasite Rickettsiella grylli were more likely to survive when the host was exposed to warmer temperatures. Crickets infected with R. grylli increased their preferred temperature from 26 C to 32 C. In A. domesticus, behavioral fever may be a specific response induced by relatively few pathogens and parasites. Behavioral fever in insects may differ in this respect from fever in mammals that can be elicited by a wide variety of parasites and pathogens.

Competition between immune function and lipid transport for the protein apolipophorin III leads to stress-induced immunosuppression in crickets.

Intense physical activity results in transient immunosuppression in a wide range of animals. We tested the hypothesis that competition between immune function and lipid transport for the protein apolipophorin III (apoLpIII) can cause transient immunosuppression in crickets. Both flying, an energetically demanding behavior, and an immune challenge reduced the amount of monomeric (free) apoLpIII in the hemolymph of crickets. Because both immune function and flying depleted free apoLpIII, these two phenomena could be in competition for this protein. We showed that immune function was sensitive to the amount of free apoLpIII in the hemolymph. Reducing the amount of free apoLpIII in the hemolymph using adipokinetic hormone produced immunosuppression. Increasing apoLpIII levels after flight by pre-loading animals with trehalose reduced immunosuppression. Increasing post-flight apoLpIII levels by injecting purified apoLpIII also reduced flight-induced immunosuppression. These results show that competition between lipid transport and immune function for the same protein can produce transient immunosuppression after flight-or-fight behavior. Intertwined physiological systems can produce unexpected trade-offs.

"Central arousal" and sexual responsiveness in the snail, Helix aspersa.

In molluscs, a "central arousal" system is thought to positively modulate both an animal's level of activity and its behavioral responsiveness. This hypothesis is examined in Helix aspersa by testing the relationships between activity, feeding, and sexual behavior. Activity, feeding, and mating exhibit parallel daily rhythms. Snails are most active, and eat and mate most frequently, during scotophase and the first 3 h of photophase. Handling and injections of serotonin (5 x 10(-7) m/kg body wt) increase general activity. Inducing activity during late photophase increases food consumption, but it does not induce sexual activity. Moreover, serotonin and handling have no effect on sexual arousal; treated snails show no increase in genital eversion stage and require the same length of courtship as controls. If snails are sexually deprived, increasing activity in late photophase does increase copulation frequency, but snails do not copulate more frequently than do unhandled snails tested during early photophase. These results suggest that while manipulations thought to increase central arousal do increase activity and responsiveness to food stimuli, they do not directly affect sexual proclivity or sexual arousal. Moreover, the relationship between feeding and sexual behavior differs between Helix and Aplysia. This difference between the two species may be related to the differences in their reproductive strategies.

Dissociation of sexual arousal and sexual proclivity in the garden snail, Helix aspersa.

Sexual arousal (intensity of courtship) and sexual proclivity (tendency to court) in Helix aspersa can be reliably measured using externally observable correlates. Snails with sexual proclivity are significantly more likely to turn toward an anesthetized conspecific after contacting it than are sexually unreceptive snails. Sexual arousal can be inferred from the stage of a snail's genital eversion, which appears only during courtship. The higher the stage of the eversion, the shorter the time required to complete introductory courtship behavior and the higher the rate of successful copulation, the fewer the number of breaks and pauses during courtship, and the longer the time a snail will spend in contact with an anesthetized conspecific. Sexual proclivity has no effect on feeding or locomotory behavior; however, sexual arousal inhibits feeding and increases locomotor activity. Snails that were allowed daily contact with conspecifics required less time to complete introductory courtship behavior relative to snails that were isolated from conspecifics for 1 week. This suggests that daily contact increases sexual arousal. A greater percentage of isolated snails exhibited courtship behavior than did snails which had experienced daily conspecific contact. This suggests that isolation increases sexual proclivity. These differences indicate that sexual arousal is not merely due to an increase in sexual proclivity.

The interactions of courtship, feeding, and locomotion in the behavioral hierarchy of the snail Helix aspersa.

Starvation both increased the number of snails that exhibited feeding (i.e., increased feeding proclivity) and decreased the latencies of response to a food stimulus (i.e., increased food arousal). Feeding inhibited locomotion, although starvation itself had no effect. Sexual arousal (the intensity of courtship) increased locomotion, but only in the absence of a mating partner. Sexual proclivity (the likelihood of a snail to court with conspecifics) had no effect on locomotion. In general, proclivity and arousal had different effects on behavior. Food deprivation did not alter the preference of sexually aroused snails for sexual stimuli over food stimuli Both starved and fed courting pairs responded to a food stimulus only during periods of low sexual arousal, although when not sexually aroused, starved snails usually increased the amount of time they spent in contact with a food stimulus. These results suggest that courtship can suppress feeding and that the expression of feeding behavior is dependent upon the occurrence of low levels of sexual arousal (time sharing).

The role of neurohormonal octopamine during 'fight or flight' behaviour in the field cricket Gryllus bimaculatus.

Octopamine has been called the 'fight or flight' hormone of insects. We tested this hypothesis by measuring octopamine levels in the haemolymph of field crickets after fighting, flying, courting and escape behaviours. Octopamine levels in the cricket Gryllus bimaculatus increased during aggressive (agonistic) behaviour from baseline levels of 4.5 +/- 2.1 pg microliters-1 haemolymph to 24.3 +/- 15.2 pg microliters-1 haemolymph, regardless of whether the cricket won or lost the encounter. Octopamine levels also increased after 5 min of flying (to 44.6 +/- 22.3 pg microliters-1) and during courtship. However, crickets did not exhibit an increase in their haemolymph octopamine levels after performing an escape run. Therefore, neurohormonal octopamine shows some, but not all, of the characteristics that would be expected if it were a component of a nonspecific 'arousal' system. Rather, octopamine may be released as a neurohormone to prepare the animal for a period of extended activity or to assist the animal in recovering from a period of increased energy demand. Antennal contact with conspecifics may provide a sensory cue that results in the release of octopamine into the haemolymph.

Segmental specialization of neuronal connectivity in the leech.

1. Every segmental ganglion of the leech Hirudo medicinalis contains two serotonergic Retzius cells. However, Retzius cells in the two segmental ganglia associated with reproductive function are morphologically distinct from Retzius cells elsewhere. This suggested that these Retzius cells might be physiologically distinct as well. 2. The degree of electrical coupling between Retzius cells distinguishes the reproductive Retzius cells; all Retzius cells are coupled in a non-rectifying manner, but reproductive Retzius cells are less strongly coupled. 3. Retzius cells in standard ganglia depolarize following swim motor pattern initiation or mechanosensory stimulation while Retzius cells in reproductive ganglia either do not respond or hyperpolarize. 4. In standard Retzius cells the depolarizing response caused by pressure mechanosensory neurons has fixed latency and one-to-one correspondence between the mechanosensory neuron action potentials and Retzius cell EPSPs. However, the latency is longer than for most known monosynaptic connections in the leech. 5. Raising the concentration of divalent cations in the bathing solution to increase thresholds abolishes the mechanosensory neuron-evoked EPSP in standard Retzius cells. This suggests that generation of action potentials in an interneuron is required for production of the EPSP, and therefore that the pathway from mechanosensory neuron to Retzius cell is polysynaptic. 6. P cells in reproductive segments have opposite effects on reproductive Retzius cells and standard Retzius cells in adjacent ganglia. Thus the difference in the pathway from P to Retzius is not localized specifically in the P cell, but elsewhere in the pathway, possibly in the type of receptor expressed by the Retzius cells.