Neurosecretory protein synthesis in relation to starvation and feeding in adult male Locusta migratoria

An immunological approach has been used to assess the effects of starvation and subsequent feeding upon the synthesis of neurosecretory protein in adult male Locusta migratoria. Starvation (for 5 days) did not reduce the incorporation of radioactive cystine into neurosecretory protein. However, the rate of neurosecretory transport in starved insects was approximately half that in normal fed insects.Within 1 hr after the start of feeding the rate of neurosecretory protein synthesis more than doubled and the rate at which newly synthesized protein was transported to the CC increased approximately three-fold. The incorporation of cystine into specific (i.e. neurosecretory) protein was always higher, even in starved insects, than into the non-specific proteins (i.e. proteins other than those from the A-cells) of the brain. This difference was maximal at 4 hr after feeding when the incorporation into specific protein was more than 5 times that into non-specific protein.     

SCHWANN CELL PROLIFERATION IN DEVELOPING MOUSE SCIATIC NERVE : A Radioautographic Study

Proliferation of Schwann cells in neonatal mouse sciatic nerve was studied radioautographically in 1-µ glycol methacrylate sections. 28 mice were injected with thymidine-³H, 4 µc/g, 48 hr after birth, and were killed serially over the next 4 days. For the cell cycle following injection, the generation time was approximately 24 hr as determined by grain-count halving data; the duration of synthesis phase was 8 hr as determined from a curve constructed from the per cent of mitotic figures containing label; and the labeling index was 9% at 2 hr after injection. With these estimates, the per cent of Schwann cells proliferating was calculated to be 27%. In addition, roughly 25% of dividing cells appeared to cease division during the cell cycle under study. The relationship of these findings to other events during maturation of nerve is discussed.     

Plastic responses of survival and fertility following heat stress in pupal and adult Drosophila virilis

The impact of rising global temperatures on survival and reproduction is putting many species at risk of extinction. In particular, it has recently been shown that thermal effects on reproduction, especially limits to male fertility, can underpin species distributions in insects. However, the physiological factors influencing fertility at high temperatures are poorly understood. Key factors that affect somatic thermal tolerance such as hardening, the ability to phenotypically increase thermal tolerance after a mild heat shock, and the differential impact of temperature on different life stages are largely unexplored for thermal fertility tolerance. Here, we examine the impact of high temperatures on male fertility in the cosmopolitan fruit fly Drosophila virilis. We first determined whether temperature stress at either the pupal or adult life history stage impacts fertility. We then tested the capacity for heat‐hardening to mitigate heat‐induced sterility. We found that thermal stress reduces fertility in different ways in pupae and adults. Pupal heat stress delays sexual maturity, whereas males heated as adults can reproduce initially following heat stress, but become sterile within seven days. We also found evidence that while heat‐hardening in D. virilis can improve high temperature survival, there is no significant protective impact of this same hardening treatment on fertility. These results suggest that males may be unable to prevent the costs of high temperature stress on fertility through heat‐hardening, which limits a species’ ability to quickly and effectively reduce fertility loss in the face of short‐term high temperature events.     

The role of antennae in the thermopreference and biting response of haematophagous bugs

Insects sense thermal cues mainly through thermoreceptors located in the antenna. To analyse the impact of antennectomy on the thermal behaviour of the haematophagous bug Triatomainfestans, we studied the distribution of intact and antennectomised bugs in an experimental arena where a temperature gradient was established, as well as the biting response of insects with and without antennae to objects at the temperature of a potential host. Antennectomy did not abolish thermopreference, but modified the temperature at which the insects preferred to stay. In the arena, antennectomised insects chose to remain at a higher temperature (ca. 3 °C higher in unfed bugs), and exhibited a larger dispersion around that preferred temperature, than intact bugs. In addition, ablated insects temporarily lost their ability to bite an object at the temperature of a potential host, but that ability was gradually recovered after the fifth day post-antennectomy. Results presented here show that thermoreceptors other than those located on the antennae can also guide thermal behaviours. We conclude that the function of antennal thermoreceptors can be taken over by other receptors located in different regions of the body. Those receptors have a different sensitivity and confer the insects with a different responsiveness.     

Development ofLydina polidoides [Diptera: Tachinidae] in relation to its hostChrysoteuchia topiaria [Lepidoptera: Pyralidae]

Lydina polidoides (Townsend) andChrysoteuchia topiaria Zeller were studied in the laboratory. The rate of development ofC. topiaria from eclosion to the diapausing prepupae was 55 days when larvae were exposed to long daylenghts (16 hr light, 8 hr dark) and 43 days when exposed to short daylengths (12 hr light 12 hr dark.) The same insects exposed to long days emerged as adults in 124 days, whereas those exposed to short days emerged in 156 days because short days prepared the prepupae for diapause but failed to maintain the diapause. When caterpillars (prepupae) ofC. topiaria parasitized by young larvae ofL. polidoides and non-parasitized caterpillars were removed from the field in January and exposed to long days at 21°C,C. topiaria emerged as adults in an average 55 days andL. polidoides in 106 days.Lydina polidoides is therefore univoltine andC. topiaria is probably the primary host of this Tachinid parasite.     

Thermosensation and the TRPV channel in Rhodnius prolixus

The thermal sense of triatomine bugs, vectors of Chagas disease, is unique among insects. Not only do these bugs exhibit the highest sensitivity to heat known in any animal up to date, but they can also perceive the infrared radiation emitted by the body of their warm-blooded hosts. The sensory basis of this capacity has just started to be unravelled. To shed additional light on our understanding of thermosensation, we initiated an analysis of the genetic basis of the thermal sense in Rhodnius prolixus. We tested the hypothesis that a TRPV (transient receptor potential vanilloid) channel receptor is involved in the evaluation of heat in this species. Two different approaches were adopted. Initially, we analysed the expression of a TRPV candidate for this function, i.e., RproIav, in different tissues. Subsequently, we tested the effects of capsaicin and capsazepine, two molecules known to interact with mammal TRPV1, using three different behavioural protocols for evaluating thermal responses: (1) proboscis extension response (PER), (2) thermopreference in a temperature gradient and (3) spatial learning in an operant conditioning context. Bioinformatic analyses confirmed that the characteristic features typical of the TRPV channel subfamily are found in the RproIav protein sequence. Molecular analysis showed that RproIav is expressed in R. prolixus, not only in the antennae, but also in other body structures bearing sensory organs. Behavioural experiments consistently revealed that capsaicin treated insects are less responsive to heat stimuli and prefer lower temperatures than non-treated insects, and that they fail to orient in space. Conversely, capsazepine induces the opposite behaviours. The latter data suggest that triatomine thermoreception is based on the activation of a TRP channel, with a similar mechanism to that described for mammal TRPV1. The expression of RproIav in diverse sensory structures suggests that this receptor channel is potentially involved in bug thermoreception. This constitutes solid evidence that thermosensation could be based on the activation of TRP receptors that are expressed in different tissues in R. prolixus. Whether RproIav channel is a potential target for the compounds tested and whether it mediates the observed effects on behaviour still deserves to be confirmed by further research.     

Time-course for attainment and reversal of acclimation to constant temperature in two Ceratitis species

Acclimation in the thermal tolerance range of insects occurs when they are exposed to novel temperatures in the laboratory. In contrast to the large number of studies that have tested for the ability of insects to acclimate, relatively few have sought to determine the time-course for attainment and reversal of thermal acclimation. In this study the time required for the Mediterranean fruit fly, Ceratitis capitata Wiedemann, and the Natal fruit fly, Ceratitis rosa Karsch, to acclimate to a range of constant temperatures was tested by determining the chill-coma recovery time and heat knock-down time of flies that had been exposed to novel benign temperatures for different durations. The time required for reversal of acclimation for both Ceratitis species was also determined after flies had been returned to the control temperature. Acclimation to 31 °C for only one day significantly improved the heat knock-down time of C. capitata, but also led to slower recovery from chill-coma. Heat knock-down time indicated that acclimation was achieved after only one day in C. rosa, but it took three days for C. rosa to exhibit a significant acclimation response to a novel temperature of 33 °C when measured using chill-coma recovery time. Reversal of acclimation after return to initial temperature conditions was achieved after only one day in both C. capitata and C. rosa. Adult C. capitata held at 31.5 °C initially exhibited improved heat knock-down times but after 9 days the heat knock-down time of these flies had declined to levels not significantly different from that of control flies held at the baseline temperature of 24 °C. In both Ceratitis species, heat knock-down time declined with age whereas chill-coma recovery time increased with age, indicating an increased susceptibility to high and low temperatures, respectively.     

Getting to the core: Internal body temperatures help reveal the ecological function and thermal implications of the lions’ mane

It has been proposed that there is a thermal cost of the mane to male lions, potentially leading to increased body surface temperatures (T_s), increased sperm abnormalities, and to lower food intake during hot summer months. To test whether a mane imposes thermal costs on males, we measured core body temperature (T_b) continuously for approximately 1 year in 18 free‐living lions. There was no difference in the 24‐hr maximum T_b of males (n = 12) and females (n = 6), and males had a 24‐hr mean T_b that was 0.2 ± 0.1°C lower than females after correcting for seasonal effects. Although feeding on a particular day increased 24‐hr mean and 24‐hr maximum T_b, this phenomenon was true of both male and female lions, and females had higher 24‐hr mean and 24‐hr maximum T_b than males, on both days when lions did not feed, and on days when lions did feed. Twenty‐four‐hour T_b was not influenced by mane length or color, and 24‐hr mean T_b was negatively correlated with mane length. These data contradict the suggestion that there exists a thermal cost to male lions in possessing a long dark mane, but do not preclude the possibility that males compensate for a mane with increased heat loss. The increased insulation caused by a mane does not necessarily have to impair heat loss by males, which in hot environments is primarily through respiratory evaporative cooling, nor does in necessarily lead to increased heat gain, as lions are nocturnal and seek shade during the day. The mane may even act as a heat shield by increasing insulation. However, dominant male lions frequent water points more than twice as often as females, raising the possibility that male lions are increasing water uptake to facilitate increased evaporative cooling. The question of whether male lions with manes compensate for a thermal cost to the mane remains unresolved, but male lions with access to water do not have higher T_b than females or males with smaller manes.     

Revisiting Trypanosoma rangeli Transmission Involving Susceptible and Non-Susceptible Hosts

Trypanosoma rangeli infects several triatomine and mammal species in South America. Its transmission is known to occur when a healthy insect feeds on an infected mammal or when an infected insect bites a healthy mammal. In the present study we evaluated the classic way of T. rangeli transmission started by the bite of a single infected triatomine, as well as alternative ways of circulation of this parasite among invertebrate hosts. The number of metacyclic trypomastigotes eliminated from salivary glands during a blood meal was quantified for unfed and recently fed nymphs. The quantification showed that ~50,000 parasites can be liberated during a single blood meal. The transmission of T. rangeli from mice to R. prolixus was evaluated using infections started through the bite of a single infected nymph. The mice that served as the blood source for single infected nymphs showed a high percentage of infection and efficiently transmitted the infection to new insects. Parasites were recovered by xenodiagnosis in insects fed on mice with infections that lasted approximately four months. Hemolymphagy and co-feeding were tested to evaluate insect-insect T. rangeli transmission. T. rangeli was not transmitted during hemolymphagy. However, insects that had co-fed on mice with infected conspecifics exhibited infection rates of approximately 80%. Surprisingly, 16% of the recipient nymphs became infected when pigeons were used as hosts. Our results show that T. rangeli is efficiently transmitted between the evaluated hosts. Not only are the insect-mouse-insect transmission rates high, but parasites can also be transmitted between insects while co-feeding on a living host. We show for the first time that birds can be part of the T. rangeli transmission cycle as we proved that insect-insect transmission is feasible during a co-feeding on these hosts.     

Flow increases tolerance of heat and hypoxia of an aquatic insect

Recent experiments support the idea that upper thermal limits of aquatic insects arise, at least in part, from a lack of sufficient oxygen: rising temperatures typically stimulate metabolic demand for oxygen more than they increase rates of oxygen supply from the environment. Consequently, factors influencing oxygen supply, like water flow, should also affect thermal and hypoxia tolerance. We tested this hypothesis by measuring the effects of experimentally manipulated flows on the heat and hypoxia tolerance of aquatic nymphs of the giant salmonfly (Plecoptera: Pteronarcys californica), a common stonefly in western North America. As predicted, stoneflies in flowing water (10 cm s⁻¹) tolerated water that was approximately 4°C warmer and that contained approximately 15% less oxygen than did those in standing water. Our results imply that the impacts of climate change on streamflow, such as changes in patterns of precipitation and decreased snowpack, will magnify the threats to aquatic insects from warmer water temperatures and lower oxygen levels.     

The initiation of melanogenesis in the chick retinal pigment epithelium.

Observations have been made on chick pigment retinal epithelium between 2 and 5 days of development. 2-Thiouracil has been demonstrated to be an effective agent for measuring the rate of melanin synthesis.Using [³H]thymidine and colcimid, we have found that the cells undergo a marked withdrawal from the cell cycle between 3 and 3.5 days of incubation in ovo, indicating that a majority of the population is synchronized. This withdrawal is followed, approximately 24 hr later, by a rapid rise in melanin synthesis from the basal level which first appears at approximately 3 days.5-Bromodeoxyuridine (BUdR) has been used to determine the time at which melanin synthesis is initiated. When BUdR is administered as early as 2 days in ovo, it is incapable of blocking the appearance of basal levels of melanin even though the cells divide at least three times in the presence of this thymidine analog. However, BUdR is capable of delaying the rapid rise in the rate of melanin synthesis first observed at 4.5 days. This delay has been found to correlate, using [³H]BUdR, with a delay in the withdrawal of the cells from the division cycle.In pursuing the idea of a correlation between withdrawal and the rapid increase of melanin formation, 5-fluorodeoxyuridine (FUdR) was used. Histological and biochemical evidence suggests that those cells which have been prevented from dividing by FUdR increase their rate of melanin synthesis to the high level of the postmitotic control cells described above.Therefore, it seems that (1), in light of work done by others, the initial decision to make melanin is made prior to 2 days in ovo, and (2) the mechanism by which cells shift their synthetic capabilities to high levels of melanin production is withdrawal mediated.     

Control of the circadian rhythm of locomotor activity in the house cricket

A detailed analysis was made of the locomotor activity of Acheta domesticus under conditions of 12 hr light and 12 hr darkness (LD 12 : 12) and of continuous darkness (DD). Under LD 12 : 12 it was found that there are three types of insects: (1) those beginning the period of increased locomotor activity immediately after darkness falls, (2) considerably before this time, and (3) considerably after this time. Under DD conditions the greater amount of the insects have a free-running rhythm shorter than 24 hr, while only a small percentage have a rhythm of more than 24 hr.Destruction of the neurosecretory cells of the pars intercerebralis by means of radio waves leads to the formation of hyperactivity and loss of locomotor activity rhythm when more than half of these cells are destroyed.Injection of reserpine into the insect's haemolymph with doses of 10 μg/g of body weight results in a reduction in locomotor activity and produces arrhythmicity for 2 to 3 days under LD 12 : 12 conditions. Under DD conditions, however, this same dose results in a total and irretrievable loss of free-running rhythm. Histological studies of the brain of crickets following injection of reserpine show a large degree of accumulation of neurosecretion in the cells of the pars intercerebralis as compared with control insects.A hypothesis is put forward as to the way in which the brain centres regulating locomotor rhythm act in crickets.     

Thymidine metabolism and the monitoring of DNA synthesis in insects

The thymidine degradation pathway established for other organisms is confirmed in insects. When ³H-TdR is used as a marker for DNA synthesis in developing silkmoths, some is incorporated into DNA and some degraded to compounds not incorporated into DNA. After a single injection, ³H-TdR is rapidly cleared from haemolymph and other tissue, resulting in, at most, a 4 hr pulse. In wing tissue, detection of DNA synthesis is possible for a maximum of 4 hr after injection of precursor and for 6 hr in vitro. Continuous monitoring of DNA synthesis can be attained by perfusion, which maintains high levels of circulating ³H-TdR.     

Ovarian and fat body protein concentrations in Ips sexdentatus (Coleoptera: Scolytidae) parasitized by nematodes

The protein concentrations of fat body and ovaries in Ips sexdentatus either uninfected or infected by Parasitaphelenchus sp., P. sexdentati, or Contortylenchus diplogaster were measured at various stages of insect development, from preswarming maturation to the first oviposition (24 hr after mating). Weight variations of the fat body and ovaries in insects infected by C. diplogaster show the same evolution as those observed in uninfected insects, but at a much lower level. Fat body proteins in uninfected insects reach their minimum level during swarming, but they remain fairly constant throughout the maturation of the first egg. After dropping shortly after swarming, the ovarian protein level in such insects increases in two stages during ovarian maturation. The first stage, which corresponds to a slow protein incorporation, takes place during the first 12 hr after mating. During the second stage, i.e., beyond 12 hr, a significant level of proteins is rapidly incorporated into the ovaries. In insects infected by Parasitaphelenchus fat body proteins are reduced and protein incorporation into the ovaries is reduced; Parasitaphelenchus would thus affect at least some proteins required for ovarian maturation in their host. Fat body protein levels are even more affected by C. diplogaster than by Parasitaphelenchus, while incorporation into ovaries seems to be less affected in spite of slower ovarian growth. C. diplogaster might thus essentially act both upon proteins which are not required for the ovarian maturation of their host and upon nonproteinaceous substances that are required for such maturation. Results are discussed in relation to the possible mode of action of parasitic nematodes.     

Thermal acclimation in a complex life cycle: The effects of larval and adult thermal conditions on metabolic rate and heat resistance in Culex pipiens (Diptera: Culicidae)

It has now been well established that insects can respond to variation in their environment via acclimation, yet the extent of the response varies among populations and environmental characteristics. One under-investigated theme which may contribute to this variation concerns acclimation effects across the life cycle. The present study explores how acclimation in the larval stage of Culex pipiens affects thermal relations in the adult stage. Mosquitoes were reared in a full factorial design at 18 or 26 °C as larvae and adults, then critical thermal maxima (CT_max) and metabolic rate–temperature relationships (MR–T) were determined for all 4 treatments. CT_max was positively affected by both larval and adult acclimation treatments. MR–T slope was significantly affected only by adult treatment: warm acclimated adults had on average shallower slopes and higher y-intercepts than cool acclimated ones. These results demonstrate that larval acclimation effects can alter adult phenotypes in a species whose life cycle includes two drastically different environments, an aquatic and a terrestrial stage. Studying insects with complex life cycles, especially those with aquatic or subterranean larval stages, can provide valuable information on the effects of thermal variability and predictability on phenotypic plasticity.     

Composition,synthetic and cytolytic activities of Galleria mellonella silk glands during the last-larval instar under the action of juvenile hormone

Within the first 48 hr of the last-larval instar of Galleria mellonella the silk glands grow but silk production is restrained. This ‘preparatory phase’ of the glands is probably maintained by juvenile hormone. Silk production and accumulation are stimulated in the ‘accumulation phase’ between 60 and 132 hr by unknown factors in the absence of juvenile hormone. The rate of RNA synthesis culminates at 84 hr but the RNA content increases until the end of cocoon spinning at 144 hr. In the following ‘regression phase’ (144–160 hr), when the glands exhibit high activities of acid and alkaline DN-ases and of acid phosphatase, the RNA and protein contents rapidly decrease, but that of DNA remains high. This phase is typical of moulting insects, is independent of juvenile hormone, and seems to be caused either by an increase in ecdysteroids or by lack of nutrients. The following ‘degeneration phase’ occurs when the surge of ecdysteroids terminates the larval-pupal transformation. Disintegration of silk glands by autolysis and phagocytosis is completed after pupal ecdysis (180 hr). Treatment of larvae with a juvenoid (ZR 512) at 48 or 132 hr in the last instar dramatically alter the composition, synthetic and cytolytic activities of silk glands. At the next ecdysis the glands attain a state very similar to that of the preparatory phase. They are capable of intensive silk production and completion of developmental cycle when the supernumerary larvae prepare for pupation. The results indicate that juvenile hormone can reverse the development of the silk glands.     

Phenoptosis in arthropods and immortality of social insects

In general, there are no drastic differences in phenoptosis patterns in plant and animal organisms. However, there are some specific features characteristic for insects and other arthropods: 1) their development includes metamorphosis with different biochemical laws at consecutive developmental stages; 2) arthropods can reduce or stop development and aging when in a state of diapause or temporal cold immobility; 3) their life cycle often correlates with seasonal changes of surroundings; 4) polymorphism is widespread — conspecifics differ by their lifespans and phenoptosis features; 5) lifespan-related sexual dimorphism is common; 6) significant situational plasticity of life cycle organization is an important feature; for example, the German wasp (Paravespula germanica) is obligatorily univoltine in the temperate zone, while in tropical regions its lifespan increases and leads to repeated reproduction; 7) life cycles of closely related species may differ significantly, for example, in contrast to German wasp, some tropical hornets (Vespa) have only one reproduction period. Surprisingly, many insect species have been shown to be subjected to gradual aging and phenoptosis, like the highest mammals. However, queens of social insects and some long-lived arachnids can apparently be considered non-aging organisms. In some species, lifespan is limited to one season, while others live much longer or shorter. Cases of one-time reproduction are rather rare. Aphagia is common in insects (over 10,000 species). Cannibalism is an important mortality factor in insects as well as in spiders. In social insects, which exist only in colonies (families), the lifetime of a colony can be virtually unlimited. However, in case of some species the developmental cycle and death of a colony after its completion are predetermined. Most likely, natural selection in insects does not lengthen individual lifespan, but favors increase in reproduction efficiency based on fast succession of generations leading to increased evolvability.     

Heat shock response of Dictyostelium

In response to a shift from 22 to 30°C the relative rate of synthesis of a small number of proteins is dramatically increased in Dictyostelium discoideum. The cells neither grow nor develop at this temperature but die slowly with a half-life of 18 hr. The major protein synthesized in response to a heat shock to 30°C in either growing cells or developing cells has an apparent molecular weight of 70,000 (70K). An increase in the relative rate of synthesis of 70K can be seen as early as 20 min following heat shock. Synthesis of 70K remains high for 4 hr at 30°C and then decreases. Similar kinetics of 70K synthesis occur during recovery at 22°C following a 1-hr heat shock. RNA synthesis during the first half-hour of heat shock is essential for the high rate of 70K measured 2 hr later. By isoelectric focusing the 70K protein can be separated into two spots, one of which overlaps one of the major heat shock proteins of Drosophila melanogaster. The relative rate of synthesis of several other proteins (82K, 60K, 43K) increases less dramatically in Dictyostelium during heat shock at 30°C. A heat shock to 34°C results in rapid synthesis of these proteins but not of 70K. The relative rates of synthesis of most other proteins made at 22°C decreases, most notably that of actin. Synthesis of heat shock proteins at 30°C does not significantly affect viability at 30°C but dramatically prolongs the period of time the cells can survive at 34°C. Thus, 30°C appears to be a stasis condition for Dictyostelium which elicits a response essential for protection from lethal temperatures. The similarity of the heat shock response in Dictyostelium to that in Drosophila and vertebrate cells suggests that certain aspects of the response may be universal in eukaryotes.     

Male accessory gland substance of Melanoplus sanguinipes: An oviposition stimulant under the control of the corpus allatum

The accessory glands of male Melanoplus sanguinipes contain an oviposition stimulant. Injection of gland extracts from mature males induces oviposition in 75 per cent of capable virgin females within 24 hr. Injection of gland extracts from allatectomized males produces no stimulatory effect. Gland extracts from mature males contain two antigens which cannot be detected in gland extracts from allatectomized males. However, both antigens can be detected in gland extracts from allatectomized males 3 days after treatment with juvenile hormone.Anion-exchange chromatography of mature gland extracts yielded two fractions which, when injected into virgin females, induced oviposition in 71 per cent of capable insects within 24 hr. These two fractions are immunologically identical to the two antigens which are absent from gland extracts of allatectomized males. We suggest that synthesis of the oviposition stimulant in Melanoplus is controlled by the corpus allatum.Injection of brain extract also induces oviposition in 100 per cent of capable virgin females within 24 hr. A possible rôle for the brain in the oviposition process is discussed.