Effect of inhibitors on chloride-dependent transmural potential in the rectal wall of Schistocerca gregaria

Previous experiments in vitro revealed a transmural potential difference (PD) and a short-circuit current (Isc) across the rectal wall of Schistocerca gregaria, which were dependent on chloride ions in lumen. The present report shows that anoxia, dinitrophenol and cyanide inhibit DP and Isc, proving that the required energy derives from oxidative metabolism. Acetazolamide also inhibits DP and Isc. Ouabain, when in haemocoele, has also an inhibitory effect, not ascribable to a Na⁺K⁺ pump blocking action. It is suggested that HCO₃^? plays an important rôle in the active transport of Cl^? from lumen to haemocoele and that ouabain may in some way inhibit chloride pumping.     

Allatectomy and sexual receptivity in females of Schistocerca gregaria

The removal of the corpora allata from immature females of Schistocerca gregaria causes the insects to enter a phase of permanent defence towards courting males. Operated control females copulate often, although they do not appear to show an active form of sexual display or attraction. Implanted corpora allata fail to restore sexual receptivity or normal ovarial development in the allatectomized females due to the inactivity of the denervated glands. The observations are discussed in relation to findings in other locusts, grasshoppers, and crickets.     

The germ-free culture of desert locusts, Schistocerca gregaria

A technique is described for rearing germ-free desert locusts. Axenic insects appeared to develop and reproduce normally, therefore it is concluded that the gut bacterial flora does not contribute significantly to locust nutrition. However, it is suggested that the gut flora may benefit its host by contributing to the locust's defences against attack by pathogenic microorganisms. The results of morphometric measurements are discussed in terms of the bacterial origin of the so called phase pheromone, locustol. No evidence was found to support the hypothesis that microbial activity restricts gut carbohydrase.     

The embryonic apolyses of Schistocerca gregaria (Orthoptera)

The origin and differentiation of the ecdysial glands and the appearance of the cuticles were studied in embryos of Schistocerca gregaria at corresponding stages. The development of the cuticle was also studied in headless embryos explanted in vitro.     

The metabolism of glycerol in the locust Schistocerca gregaria during flight

The concentration of glycerol in locust haemolymph increases 10-fold during 1 hr flight but decreases rapidly when flight ceases. [¹⁴C]Glycerol is rapidly metabolized by locusts in vivo. Trehalose and diacyl glycerol are the main products to appear in the haemolymph but the proportion of diacyl glycerol is increased in flown insects or when adipokinetic hormone is injected. Trehalose and diacyl glycerol are also the main products formed when isolated fat body is incubated with [¹⁴C]glycerol. Adipokinetic hormone increases the proportion of diacyl glycerol formed.It is proposed that during flight glycerol is produced by hydrolysis of diacyl glycerol in the flight muscles. It is then transported to fat body for esterification with fatty acid produced during conversion of triacyl glycerol stores to diacyl glycerol.     

Control of melanization in first instar larvae of Schistocerca gregaria

Melanization in first-instar larvae of Schistocerca is controlled by a hormone released from neurosecretory axon terminals of the fine nerves posterior to the metathoracic ganglion. The hormone is not detectable in the haemolymph before the embryonic ecdysis but is present within seconds after the ecdysis has started. It is suggested that horizontal displacement of the embryonic cuticle is the trigger for the release of the hormone and that the prothoracic ganglion forms part of the neural pathway between the sensory input caused by ecdysis and the release of the hormone.     

Ecdysteroids in the developing eggs of the desert locust, Schistocerca gregaria

Ecdysteroid levels in the developing eggs of Schistocerca gregaria were determined, at daily intervals, using gas chromatography and electron capture detection of ecdysteroid derivatives. Ecdysone and 20-hydroxyecdysone were present both as the free ecdysteroid and as polar conjugates. Total ecdysteroids reached a maximum of 40 ng/egg with ecdysone contributing the greater part.     

Comparison between the sclerotization of adult and larval cuticle in Schistocerca gregaria

Femur cuticle from fifth instar larvae of the desert locust, Schistocerca gregaria, has been characterized with respect to composition, rate of deposition, and rate of sclerotization. The results are compared with those from adult cuticle of the same species. The protein compositions of the two types of cuticle are very similar, but the rates of deposition of both protein and chitin are different. The main difference is, however, that sclerotization is restricted to the first day after ecdysis in larval cuticle, whereas in adult cuticle sclerotization continues for at least a couple of weeks. The result is that the endocuticle remains untanned in the larvae, whereas in the adults the whole cuticle becomes tanned.     

Componental analysis of the ocellar electroretinogram of the locust, Schistocerca gregaria

Ruck's componental analysis of the ocellar electroretinogram (ERG) has been reappraised using techniques of signal averaging and waveform subtraction. Components (1), (3), and (4) can readily be isolated in the locust ocellus but component (2) as recorded in the locust ocellus is probably an artefact. Component (1), produced by the receptor cells, only contributes significantly to the total ERG at higher light intensities and it is this contribution which changes most with the degree of light and dark adaptation employed in these experiments. Component (3), the response of the second-order neurones, indicates that the majority of second-order neurones hyperpolarize on illumination of the ocellus. Component (4), the afferent activity of the second-order cells, indicates that more than one afferent axon is involved in the production of off spikes in the locust ocellus.     

Studies on the moulting hormones of the desert locust, Schistocerca gregaria

A chemical investigation of the desert locust (Schistocerca gregaria) using an isolated locust abdomen assay has led to the identification of 20-hydroxyecdysone as one of the hormones controlling moulting, but the evidence presented does not favour the prothoracic gland (PTG) as the site of its production. Preliminary information indicates two other active substances are present in higher concentration in the PTG which affect apolysis. Determination of 20-hydroxyecdysone titre at daily intervals in the fifth instar and adult show highest concentrations on the day of ecdysis. Ecdysone was not detected. Histological examination of cuticle suggests that PTG extracts cause growth in epidermal cells, rather than increased cell division.     

Some aspects of water absorption by the developing egg of Schistocerca gregaria

Schistocerca gregaria eggs absorb water over their entire surface. Water entry is facilitated by changes in the permeability of cuticular membranes covering the egg. The shell is markedly more permeable during the period of rapid water uptake and much less so on the other days. The changes in permeability are due to the differing amounts of lipoid material in the shell at different developmental periods. The absorption appears to be an active process requiring oxidative energy, although osmosis may also be partly involved. Water entry is mediated by the serosa and is not influenced by the developing embryo.     

Experimental analysis of the locomotor behaviour of Schistocerca gregaria induced by odour

The locomotor behaviour of the desert locust in response to volatile chemicals released from grass as a food plant was investigated in a wind tunnel. When bruised grass was at the windward end of the tunnel the insects responded by moving upwind towards the grass. The response was maximal for locusts starved for 4 hr or longer. Feeding just prior to an experiment inhibited the response; the degree of inhibition of the response, as estimated by the proportion of larvae responding, increased with the duration of the feeding period. All the larval instars and adults reacted in essentially the same way, but the response varied in the course of any one instar, there being more response in the first half of the instar—that is, the period preceding peak food intake—than later.     

Development of the accessory reproductive glands and genital ducts in female Schistocerca gregaria

The accessory reproductive glands of female S. gregaria are tubular extensions of the paired genital ducts, which in the mature female contain large amounts of a proteinaceous secretion used in the formation of the egg pod. In the 4th and 5th-instar female the glands are indistinguishable from the remainder of the mesodermally derived genital ducts. Towards the end of the 5th stadium, however, the accessory gland region only acquires characteristic convolutions which persist throughout the adult stages. At this time the epithelium of the entire ducts becomes reorganized into a unicellular epithelium. Only one cell type occurs throughout the length of the glands, and also in the egg calyces and lateral oviducts. The cells are inactive immediately after final ecdysis and remain in this state until the level of juvenile hormone in the haemolymph rises. The hormone acts directly on the cells triggering a rapid proliferation of organelles associated with protein secretion, and thereby increasing the volume of apical cytoplasm. Microvilli develop at the luminar plasma membrane, while irregular infoldings form at the base of the cells. As the gland matures the major organelle, the rough endoplasmic reticulum, changes from the lamellar to the vesicular form. Secretion is released into the lumen by the ‘microapocrine’ method.     

The hormonal control of accessory reproductive gland development in female Schistocerca gregaria

Allatectomy of adult female Schistocerca gregaria prevents the normal development of the accessory reproductive glands and no secretion is produced. Development of the glands can be restored by the administration of synthetic juvenile hormone and the response is dose-dependent. A continuous supply of hormone is required for maintenance of secretory activity. In the normal developmental sequence the total protein content of the glands remains constant until the time at which vitellogenesis occurs in the terminal oöcytes. As maturation proceeds there is a linear increase in protein content of the glands. The initial increase occurs as a result of cellular changes in the glands and is then followed by an increase due to an accumulation of secretion in the lumina.     

Ecdysteroid levels throughout the life cycle of the desert locust, Schistocerca gregaria

Moulting hormone levels for all stages of the life cycle of the desert locust, Schistocerca gregaria, have been determined using gas chromatography with electron capture detection of the trimethylsilylated hormones. During larval development, the major hormone detected is 20-hydroxyecdysone with smaller quantities of ecdysone present. In mature adult females the major ecdysteroid observed is a polar conjugate of ecdysone, with smaller quantities of conjugated 20-hydroxyecdysone also present. During embryonic development the pattern changes from a high proportion of conjugated ecdysone in the early stages to give more free hormone and a higher proportion of 20-hydroxyecdysone in later stages. The highest titre of 20-hydroxyecdysone found in this insect is during the 5th larval instar. Maximal levels of ecdysteroid per insect are found in mature females just before oviposition, while the highest level of ecdysteroid per g of tissue is found in the eggs.     

The need for high levels of phenylalanine in the diet of Schistocerca gregaria nymphs

It has been demonstrated that an apparently good, nutrient-rich plant can be relatively short of phenylalanine for fifth-instar nymphs of Schistocerca gregaria. Supplementary phenylalanine invariably increased the efficiency of conversion of ingested food to body substance, showing that this was the limiting nutrient. Relatively large quantities of phenylalanine are required for cuticle protein over a few days in the first half of the instar, which is when this amino acid is at a premium.     

The pattern of ecdysone levels during development in the desert locust, Schistocerca gregaria

A simple chemical assay, sensitive to picogram amounts (10^?12 g) of ecdysones in crude biological materials is described. By a simple, rapid purification procedure followed by conversion to their trimethylsilyl ethers, ecdysones are determined quantitatively by gas chromatography with an electron capture detector. The method has been applied to the determination of ecdysones in the fourth and fifth instars of the desert locust. 20-Hydroxyecdysone is always present in higher concentration than ecdysone and the former rises to a sharp maximum 1 to 2 days before ecdysis.     

A re-investigation on the ecdysteroids during embryogenesis in the desert locust, Schistocerca gregaria

Comparison of gas chromatography and high-pressure liquid chromatography methods for determination of ecdysteroids in insect eggs revealed that published values for Schistocerca gregaria were very low. The error has been traced to incomplete hydrolysis of conjugates. Revised values for the levels of ecdysone, 20-hydroxy-ecdysone and 2-deoxy-ecdysone are given here. The same general pattern of falling ecdysteroid titre during early stages of embryo development followed by an increase and a final decrease is observed. Experiments have been made on the effect of phosphate, ionic strength and a specific β-glucuronidase competitive inhibitor on the hydrolysis of the polar ecdysteroid conjugates, in an attempt to provide more evidence on the chemical nature of these compounds.     

Juvenile hormone activity in larvae and adult females of the locust, Schistocerca gregaria

The isolation and purification of fractions with juvenile hormone activity from whole animal extracts of larvae, and from extracts of haemolymph from larvae and adults is described. Using the Galleria bioassay three such fractions were isolated from both third and fourth instar hoppers and from adult females. The chromatographic behaviour of these fractions indicates that one contains JH III, but the other two contain unknown juvenilizing compounds.     

Changes in the first instar cuticle of Schistocerca gregaria before and associated with hatching

Size increases in the first instar cuticle are described in the pharate stage, at hatching and ecdysis, and in the hour following ecdysis. The ways in which the cuticle expands are described. Changes in colour and the mechanical properties of cuticle are described and related to changes in the different protein fractions extracted from the cuticle. A blood-borne factor is shown to affect darkening in the first instar.