Effect of β-cyano-l-alanine on the water balance of Locusta migratoria

Administration of β-cyano-l-alanine (BCNA) to adult locusts results in a significant decrease in haemolymph volume within one day. Dehydration becomes acute within 2 to 3 days and causes death within 5 days. This syndrome is attributed to impaired ability of BCNA-treated locusts to reabsorb water into the haemolymph from the rectal lumen. Extracts of brains and last abdominal ganglia, or haemolymph of water deprived locusts have no effect on rectal water resorption in BCNA-treated locusts. The initial symptoms of BCNA toxicity can be temporarily alleviated by injection of water into the haemocoel. Chronic BCNA toxicity irreversibly disrupts the control of water balance in locusts.     

The role of the glandular lobes of the corpora cardiaca during flight in Locusta

ABSTRACT. Flight performance in Locusta is reduced following severance of the major afferent nerves to the corpora cardiaca or removal of the glandular lobes of the corpora cardiaca. These operations prevent the release of adipokinetic hormone and the consequent mobilization of stored lipid. However, locusts deprived of about 90% of their glandular lobe tissue, while flying poorly, did mobilize lipid. It is suggested that the remaining glandular parenchyma cells are capable of secreting enough hormone to stimulate lipid mobilization, but that the concentration may be inadequate to encourage lipid utilization. After removal of all the glandular lobe parenchyma, the blood carbohydrate concentration was temporarily depressed. Nevertheless flight performance was equally poor, both when haemolymph carbohydrate levels were low and when they had returned to normal. After the injection of trehalose into operated control locusts and locusts deprived of their glandular lobes, flight was still markedly poorer in the operated insects, even though the injection of trehalose prevented adipokinetic hormone release in the intact locust. It seems that the poor flight performance of locusts deprived of their glandular lobes cannot be fully explained by the simple absence of adipokinetic hormone.     

Water storage and osmotic pressure influences on the water relations of a dicotyledonous desert succulent

Abstract Water storage and nocturnal increases in osmotic pressure affect the water relations of the desert succulent Ferocactus acanthodes, which was studied using an electrical circuit analog based on the anatomy and morphology of a representative individual. Transpiration rates and osmotic pressures over a 24-h period were used as input variables. The model predicted water potential, turgor pressure and water flow for various tissues. Plant capacitances, storage resistances and nocturnal increases in osmotic pressure were varied to determine their role in the water relations of this dicotyledonous succulent. Water coming from storage tissues contributed about one-third of the water transpired at night: the majority of this water came from the nonphotosynthetic, water storage parenchyma of the stem. Time lags of 4 h were predicted between maximum transpiration and maximum water uptake from the soil. Varying the capacitance of the plant caused proportional changes in osmotically driven water movement but changes in storage resistance had only minor effects. Turgor pressure in the chlorenchyma depended on osmotic pressure, but was fairly insensitive to doubling or halving of the capacitance or storage resistance of the plant. Water uptake from the soil was only slightly affected by osmotic pressure changes in the chlorenchyma. For this stem succulent, the movement of water from the chlorenchyma to the xylem and the internal redistribution of water among stem tissues were dominated by nocturnal changes in chlorenchyma osmotic pressure, not by transpiration.     

Water balance in Collembola and its relation to habitat selection: Water content,haemolymph osmotic pressure and transpiration during an instar

In the two collembolan species Orchesella cincta and Tomocerus minor the water content, haemolymph osmotic pressure and transpiration rate fluctuate with the feeding rhythm during each instar. The changes in water content, however, are due to changes in dry weight, because the absolute water weight stays constant during the instar.The intake of food is probably the cause of the increase in haemolymph osmotic pressure. Increase of osmotically active substances in the blood and/or blood volume reduction may be responsible for the rise in osmotic pressure. This change in osmotic pressure in turn may affect the responsiveness of the animals to water as well as their feeding behaviour.Changes in the epicuticle and in epidermal cell membranes may cause changes in the rate of transpiration. The high rate observed during ecdysis and during the mid-instar may explain the behaviour of the animals in varied water conditions.Dehydration during the instar causes an equivalent rise in osmotic pressure for both Tomocerus minor and Orchesella cincta. The water loss appears to involve the haemolymph. The physiological state of the animal does not influence the rise in osmotic pressure. There are no signs of any osmoregulation in the two species.     

Changes in haemolymph octopamine levels associated with food deprivation in the locust, Schistocerca gregaria

ABSTRACT. Levels of octopamine in the haemolymph and locomotor activity have been measured over 24 h in fed and food-deprived adult male locusts, Schistocerca gregaria (Forskål). Locusts are predominantly diurnal, the locomotor activity of individuals with continuous access to food was higher during the photophase. The basal level of octopamine in haemolymph sampled during the first quarter of the light cycle was 7.8±1.1 pg/μl (51 nm). Depriving locusts of food at the start of the photophase for 9 h caused an increase in both the speed of movement and total activity compared to fed individuals. There was a corresponding increase in the haemolymph octopamine concentration which continued for as long as the insects were unable to feed, reaching a concentration of 20.7 ± 3.7 pg/μl (135 nM). While declining, the levels of octopamine remained higher in fed locusts for up to 4 h after replacing the food. The results are discussed in relation to the regulation of feeding behaviour and a proposed neurohormonal role of octopamine in insects.     

The effect of Metarhizium anisopliae var acridum on haemolymph energy reserves and flight capability in the desert locust, Schistocerca gregaria

Adult desert locusts, Schistocerca gregaria , 3 days after inoculation with the entomopathogenic fungus Metarhizium anisopliae var acridum , had significantly less carbohydrate and lipid in the haemolymph than controls. This was not due to reduced food intake as 3 days of complete starvation had no effect on haemolymph titres of energy reserves in controls. Furthermore injection of an extract of the corpora cardiaca (the source of adipokinetic hormone, AKH) caused a large significant increase in haemolymph lipid in mycosed locusts, indicating the availability of significant quantities of lipid in the fat body, the target for AKH. Haemolymph carbohydrate declined significantly during tethered flight of control locusts but not in mycosed individuals. An injected supplement of trehalose significantly boosted flight performance of mycosed insects but not controls. The results are discussed in the light of the hypothesis that the poor flight capability of mycosed locusts is due in part to a fungus-induced reduction in mobile energy reserves.     

Feeding causes the appearance of a factor in the haemolymph that stimulates protein synthesis

Soon after a locust (Locusta migratoria) begins to feed, an increase in protein synthesis can be detected in the animal. Isolation of fat body shows that this tissue synthesizes protein at a faster rate in recently fed animals than it does in fasting insects. Fasting locusts injected with haemolymph from fed insects increased protein synthesis when compared with locusts injected with haemolymph from fasting locusts. The factor causing this increase was present in the haemolymph within 5 min of feeding. Feeding or direct contact with the food was not essential to increase protein synthesis. Exposure of fasting locusts to feeding insects was sufficient to elevate the rates of protein synthesis in the fasting animals.The increase inprotein synthesis was not a result of general excitation or an increase in the concentration of tryptophan or isoleucine in the haemolymph. Ecdysteroid titres were uniformly low during the first ten days of adult life. Gel filtration of the fed haemolymph revealed a low molecular weight fraction (about 600 daltons) which stimulated protein synthesis upon injection into fasting locusts.     

The post-prandial rest in Locusta migratoria nymphs and its hormonal regulation

Doppler radar actographs have been used to measure the fall in locomotor activity following feeding in mid-fifth instar nymphs of Locusta migratoria. Filling the crops of the insects with agar via a cannula, reduced activity in a similar manner. Haemolymph from recently-fed donors injected into insects which had been deprived of food, likewise reduced locomotor activity compared with haemolymph from deprived donors. Injections of nutrients were ineffective, but homogenates of the storage lobes of the copora cardiaca from food-deprived donors proved effective compared with similar homogenates from newly-fed donors. It is suggested that neurosecretion released from the corpora cardiaca, known to occur following crop-filling, leads to a reduction in locomotor activity.     

Some problems of homeostasis of haemolymph metabolites in the locust

Sectioning of the afferent nerves (NCCI and II) to the locust corpora cardiaca, glandular lobe removal, cardiacectomy, or removal of the median neurosecretory cells of the brain, have no long-term effect on blood lipid concentration. After removal of the glandular lobe, haemolymph carbohydrate concentration is lowered and remains significantly so from the second to the sixth day after the operation but returns to normal within 10 to 15 days. Severance of the afferent nerves to the corpora cardiaca does not, however, affect blood carbohydrate concentration. The injection of a concentrated extract of glandular lobes into locusts deprived of their glandular lobes does not elicit a hyperglycaemic effect even when blood carbohydrate levels are low. Cauterization of the median neurosecretory cells of the brain, sectioning of the NCCI and II, or removal of the glandular lobes of the corpora cardiaca have no effect on haemolymph protein.After dilution of haemolymph constituents by the injection of water, carbohydrate and protein concentrations are not rapidly restored to their initial values. The lipid concentration, however, rapidly returns to its pre-injection level due to the mobilization of 16:16, 16:18, and 18:18 diglycerides. This occurs even in glandular lobe deprived, median neurosecretory cell cauterized, or headless locusts. These diglycerides are mobilized following the injection of solutions containing lipid, carbohydrate, and/or protein, and are the same diglycerides that are released from the fat body in response to adipokinetic hormone. It is concluded that the injection of large volumes of fluid causes lipid mobilization but adipokinetic hormone does not apear to be involved, and the mechanism of blood lipid homeostasis in the resting locust is not clear.     

Heat stress is associated with disruption of ion balance in the migratory locust,Locusta migratoria

Thermal tolerance is important in determining the spatial and temporal distributions of insects but the mechanisms which determine upper thermal limits remain poorly understood. In terrestrial insects heat tolerance is unlikely to be limited by oxygen supply but in some arthropods, heat stress has been shown to cause haemolymph hyperkalaemia which is known to have detrimental effects on neuromuscular excitability. It is however unresolved if heat-induced hyperkalemia is the cause or the result of cellular heat injury. To address the putative role of heat-induced hyperkalemia we quantified changes in ion and water balance in haemolymph and muscle tissue of the migratory locust during exposure to two static temperatures clustered around the CT_max (48 °C and 50 °C). We show that heat stress caused a loss of ion balance and severe haemolymph hyperkalaemia which coincided with the onset of heat stupor. Locusts were able to maintain their haemolymph volume throughout exposure, suggesting it is unlikely that osmoregulatory failure is responsible for haemolymph hyperkalaemia. When locusts were allowed to recover from heat stupor, they recovered ion balance quickly but were still unable to function optimally after 24 h. The results therefore indicate that both the haemolymph hyperkalaemia and associated depression of muscular function (heat stupor) are secondary results of cellular heat injury and that the cause of heat stupor most be sought elsewhere.     

Some effects of amputation of the antennae on pigmentation, growth and development in the locust, Schistocerca gregaria

ABSTRACT. Removal of the antennae from Schistocerca gregaria early in the third or fourth instar resulted in fifth instar nymphs with a green haemolymph and cuticle. Antennectomized adults had a lower somatic dry weight and a smaller fat body than operated control insects. In addition, their rate of sexual development increased and some of their morphometric parameters were altered. Implantation of corpora allata into fourth instar nymphs induced a green coloration of the haemolymph after a few days. It is suggested that antennectomy of Schistocerca nymphs resulted in an increased effectiveness of the corpus allatum hormone with its concomitant effects on nymphal pigmentation, growth and development, and adult morphometries. It is further suggested that antennectomy of crowded locusts may mimic, for them, the uncrowded condition, by reducing sensory input.     

Circulating angiotensin II mediates sodium appetite in adrenalectomized rats

We investigated the role of circulating ANG II in sodium appetite after adrenalectomy. Adrenalectomized rats deprived of their main access to sodium (0.3 M NaCl) for 9 h drank 14.1 +/- 1.5 ml of the concentrated saline solution in 2 h of access. Intravenous infusion of captopril (2.5 mg/h) during the last 5 h of sodium restriction reduced sodium intake by 77 +/- 12% (n = 5) without affecting the degree of sodium depletion and hypovolemia incurred during deprivation. Functional evidence indicates that this dose of captopril blocked production of ANG II in the peripheral circulation, but not in the brain; that is, injection of ANG I into the lateral brain ventricle stimulated intake of both water and 0.3 M NaCl. Intravenous infusion of ANG II (starting 10-15 min before 0.3 M NaCl became available) in adrenalectomized, captopril-treated rats restored both sodium intake and blood pressure to values seen in rats not treated with captopril. Longer (20 h) infusions of captopril in 22-h sodium-restricted rats also blocked sodium appetite, but reduced or prevented sodium depletion. Intravenous infusion of ANG II after these long captopril infusions stimulated sodium intake, but intake was less than in controls not treated with captopril. These results indicate that most or all of the sodium appetite of adrenalectomized rats is mediated by circulating ANG II.     

Water Uptake and Protein Synthesis in Germinating Wheat Embryos under the Osmotic Stress of Polyethylene Glycol

The effect of osmotic stress on wheat-seed germination was testedby imbibition in aqueous polyethylene glycol solutions at differentconcentrations. The experiments were designed to allow blockingand the subsequent recovery of germination by 12 h or 24 h pre-imbibitionof seeds in osmoticum, followed by transfer to water. Seedswere alternatively presoaked in water for 12 or 24 h, then transferredto polyethylene-glycol solutions to study the induced blockingof germination. Water content and [³H]leucine incorporationinto embryo tissues (as a measure of in vivo protein synthesis)were determined over a 48-h imbibition period. A close relationshipwas established overall between hydration status and proteinsynthesis rate. Osmotic stress seems to have a strong influenceupon the quantitative synthesis of proteins, suggesting thatthis biochemical activity is associated with the regulationof the germination process. Triticum durum, embryo, osmotic stress, water uptake, protein synthesis     

Quantitative determination of the juvenile hormones in the haemolymph of Locusta migratoria during normal development and after implantation of corpora allata

Simultaneous quantitative determination of the three naturally occurring juvenile hormones in insects (JH-I, JH-II and JH-III) was performed on haemolymph samples of both normally developing locusts and locusts implanted with active corpora allata, using capillary gas chromatography with electron capture detection.In fourth instar female larvae, 24–48 hr after the third ecdysis, as well as in adult females, 18 days after the imaginal ecdysis, only JH-III was detected. In fifth instar female larvae JH-III was present in very low concentrations, if at all.After implantation of four pairs of corpora allata taken from young fourth instar female larvae or one pair or corpora allata taken from adult females into fifth instar female larvae 0–24 hr after ecdysis, an elevation of the JH-III titre was observed. Neither JH-I nor JH-II could be detected. The amount of JH-III, already elevated 2 hr after implantation, remained high for several days in comparison to that of control insects. On the third day after the subsequent moult the JH-III level was comparable to that of normally developing fifth instar larvae. Factors involved in the achievement of the haemolymph JH-titre are discussed.     

Ionic and osmotic regulation of the haemolymph of the dragonfly, Libellula quadrimaculata (Odonata : Libellulidae)

Larvae of the widespread dragonfly, Libellula quadrimaculata, were adapted to a series of salt solutions, and the osmotic pressure, and sodium, potassium and chloride concentrations in the haemolymph measured. The regulation of potassium is extremely efficient over the range 0–50 m-mole/l. external concentration. Above this, larvae die. Sodium and chloride are regulated to a lesser extent, the larvae being able to withstand considerable changes in the concentration of these ions in the haemolymph. However, at higher external concentrations, the haemolymph concentration of these ions is maintained below that of the external medium. The osmotic pressure is regulated in parallel with sodium concentration over most of the range tested. However, in higher salinities, the osmotic pressure of the haemolymph does not fall below that of the external medium. This is seen as a strategy to limit the amount of drinking in saline media. Overall, the osmoregulatory system of L. quadrimaculata resembles that of brackish-water insects, rather than that of the more strictly freshwater dragonflies that have been studied.     

Hindbrain serotonin and the rapid induction of sodium appetite

Both systemically administered furosemide and isoproterenol produce water intake (i.e., thirst). Curiously, however, in light of the endocrine and hemodynamic effects produced by these treatments, they are remarkably ineffective in eliciting intake of hypertonic saline solutions (i.e., operationally defined as sodium appetite). Recent work indicates that bilateral injections of the serotonin receptor antagonist methysergide into the lateral parabrachial nuclei (LPBN) markedly enhance a preexisting sodium appetite. The present studies establish that a de novo sodium appetite can be induced with LPBN-methysergide treatment under experimental conditions in which only water is typically ingested. The effects of bilateral LPBN injections of methysergide were studied on the intake of water and 0. 3 M NaCl following acute (beginning 1 h after treatment) diuretic (furosemide)-induced sodium and water depletion and following subcutaneous isoproterenol treatment. With vehicle injected into the LPBN, furosemide treatment and isoproterenol injection both caused water drinking but essentially no intake of hypertonic saline. In contrast, bilateral treatment of the LPBN with methysergide induced the intake of 0.3 M NaCl after subcutaneous furosemide and isoproterenol. Water intake induced by subcutaneous furosemide or isoproterenol was not changed by LPBN-methysergide injections. The results indicate that blockade of LPBN-serotonin receptors produces a marked intake of hypertonic NaCl (i.e., a de novo sodium appetite) after furosemide treatment as well as subcutaneous isoproterenol.     

Water flow and water storage in Agave deserti: osmotic implications of crassulacean acid metabolism

Abstract Water flow and water storage were investigated for Agave deserti, a desert succulent showing crassulacean acid metabolism (CAM). The anatomy and water relations of the peripheral chlorenchyma, where CAM occurs, and the central water-storage parenchyma were investigated for its massive leaves so that these tissues could be incorporated as discrete elements into an electrical-circuit analogue of the whole plant. The daily cycling of osmotic pressure was represented by voltage sources in series with the storage capacitors. With soil water potential and leaf transpiration rate as input variables, axial water flow through the vascular bundles and radial flows into and out of storage during the day/night cycle were determined. The predominantly nocturnal transpiration was coincident with increases in cell osmotic pressure and in titratable acid of the leaf chlorenchyma. In the outer layers of the chlorenchyma, water potential was most negative at the beginning of the night when transpiration was maximum, while the water-storage parenchyma reached its minimal water potential 9 h later. The roots plus stem contributed 7% and the leaves contributed 50% to the total water flow during maximal transpiration; peak water flow from the soil to the roots occurred at dawn and was only 58% of the maximal transpiration rate. Over each 24-h period, 39% of the water lost from the plant was derived from storage, with flow into storage occurring mainly during the daytime. Simulations showed that the acid accumulation rhythm of CAM had little impact on water uptake from the soil under the conditions employed. In the outer chlorenchyma, water potential and water flows were more sensitive to the day/night changes in transpiration than in osmotic pressure. Nevertheless, cell osmotic pressure had a large influence on turgor pressure in this tissue and determined the extent to which storage was recharged during the latter part of the night.     

The nature of the hyperglycaemic factor from the glandular lobe of the corpus cardiacum of Locusta migratoria

Extracts of the glandular lobe of the corpus cardiacum increased haemolymph carbohydrate levels in neck-ligated adults of Locusta migratoria. No change was observed in normal locusts. Synthetic AKH had no effect on haemolymph carbohydrate in neck-ligated insects. Fractionation of the glandular lobe extract by gel filtration yielded a fraction which possessed both hyperglycaemic and hyperlipaemic activity in neck-ligated insects. Crustacean red-pigment-concentrating hormone possessed hyperglycaemic activity.It is surmised that the ‘second peptide’ (AKH II) functions, at least in part, to regulate blood sugar levels.     

Nitric oxide modulates sodium taste via a cGMP-independent pathway

Insects, like other animals, require sodium chloride (NaCl) as part of their normal diet and detect it with contact chemoreceptors on the body surface. By adjusting the responsiveness of the chemosensory neurons within these receptors insects can modify the intake of salt and other nutrients, and it has been hypothesized that the responsiveness of chemosensory neurons is regulated by nitric oxide (NO). To identify potential sources of NO in the periphery, the authors applied the NO-sensitive fluorescent probe 4,5-diaminofluorescein and the universal NO synthase antibody, and found that in locusts NO is synthesized within one particular class of cells of the epidermis, the glandular cells, from where it may diffuse to neighboring chemosensory neurons. The effects of NO on chemosensory neurons were investigated by recording from contact chemoreceptors on the leg while perfusing it with drugs that interfere with NO signaling. Results showed that both endogenous and exogenous NO decreased the frequency of action potentials in chemosensory neurons in response to stimulation with NaCl by acting via a cyclic guanosine monophosphate-independent pathway. Variation of the NaCl concentration in the perfusion solution demonstrated that the synthesis of NO in glandular cells depends on the NaCl concentration in the hemolymph. By contrast NO increased the frequency of action potentials in chemosensory neurons in response to sucrose stimulation. The authors suggest that NO released from glandular cells modulates the responsiveness of chemosensory neurons to regulate NaCl intake, and hypothesize that NO may play a key role in the signaling of salt and sugars.     

Behavioural correlates of phenotypic plasticity in mouthpart chemoreceptor numbers in locusts

Rearing locusts in an impoverished chemosensory environment leads to fewer chemoreceptors developing on the mouthparts and antennae as adults but the behavioural relevance of these changes remains unknown. To address this question, locusts were reared for the final two larval stadia on either a single, nutritionally near-optimal synthetic food ('plain' pretreatment), or a diet comprising two nutritionally complementary foods containing two added flavours ('mix' pretreatment). Insects reared on the 'mix' diet had a mean 20% more chemosensilla on the maxillary palps than those fed on the 'plain' diet. Following an equilibration period, when all newly moulted adults could feed on two nutritionally complementary foods, insects were food deprived for 2 or 6 h, and then given a test meal of a single balanced food at one of two dilutions whilst their behaviour was recorded. 'Mix'-pretreated locusts had a shorter latency to feed and were more likely to reject the test food upon first contact if deprived for only 2 h; but if they did take a meal it lasted longer and contained fewer pauses. Using sensilla number as a covariate removed the statistical significance of pretreatment regime, indicating that sensilla number, or some close correlate of it, can largely account for the variation in behaviour. This suggests that sensilla numbers are behaviourally relevant; particularly where locusts are not greatly food deprived and faced with marginally acceptable foods.