Behavioral analysis of polarization vision in tethered flying locusts

For spatial navigation many insects rely on compass information derived from the polarization pattern of the sky. We demonstrate that tethered flying desert locusts (Schistocerca gregaria) show e-vector-dependent yaw-torque responses to polarized light presented from above. A slowly rotating polarizer (5.3° s^–1) induced periodic changes in yaw torque corresponding to the 180° periodicity of the stimulus. Control experiments with a rotating diffuser, a weak intensity pattern, and a stationary polarizer showed that the response is not induced by intensity gradients in the stimulus. Polarotaxis was abolished after painting the dorsal rim areas of the compound eyes black, but remained unchanged after painting the eyes except the dorsal rim areas. During rotation of the polarizer, two e-vectors (preferred and avoided e-vector) induced no turning responses: they were broadly distributed from 0 to 180° but, for a given animal, were perpendicular to each other. The data demonstrate polarization vision in the desert locust, as shown previously for bees, flies, crickets, and ants. Polarized light is perceived through the dorsal rim area of the compound eye, suggesting that polarization vision plays a role in compass navigation of the locust.     

Octopaminergic modulation of locust motor neurones

The effect of octopamine on the fast extensor and the flexor tibiae motor neurones in the locust (Schistocerca gregaria) metathoracic ganglion, and also on synaptic transmission from the fast extensor to the flexor motor neurones, was examined. Bath application or ionophoresis of octopamine depolarized and increased the excitability of the flexor tibiae motor neurones. 1 mM octopamine reduced the amplitude of the fast extensor-evoked EPSP in the slow but not the fast flexor motor neurones, whereas 10 mM octopamine could reduce the EPSP amplitude in both. Octopamine broadened the fast extensor action potential and reduced the amplitude of the afterhyperpolarization, the modulation requiring feedback resulting from movement of the tibia. Octopamine also increased the frequency of synaptic inputs onto the tibial motor neurones, and could cause rhythmic activity in the flexor motor neurones, and reciprocal activity in flexor and extensor motor neurones. Octopamine also increased the frequency of spontaneous spiking in the octopaminergic dorsal unpaired median neurones. Repetitive stimulation of unidentified dorsal unpaired median neurones could mimic some of the effects of octopamine. However, no synaptic connections were found between dorsal unpaired median neurones and the tibial motor neurones. The diverse effects of octopamine support its role in mediating arousal.     

Evolution and structural conservation of the control region of insect mitochondrial DNA

The control regions of mitochondrial DNA of two insects, Schistocerca gregaria and Chorthippus parallelus, have been isolated and sequenced. Their sizes are 752 by and 1,512 bp, respectively, with the presence of a tandem repeat in C. parallelus. (The sequences of the two repeats are highly conserved, having a homology of 97.5%.) Comparison of their nucleotide sequences revealed the presence of several conserved sequence blocks dispersed through the whole control region, showing a different evolutionary pattern of this region in these insects as compared to that in Drosophila. A highly conserved secondary structure, located in the 3 region near the small rRNA gene, has been identified. Sequences immediately flanking this hairpin structure rather than the sequences of this structure themselves are conserved between S. gregaria/C. parallelus and Drosophila, having a sequence consensus of TATA at 5 and GAA(A)T at 3. The motif G(A)nT is also present in the 3 flanking sequences of mammalian, amphibian, and fish mitochondrial L-strand replication origins and a potential plant mitochondrial second-strand-replication origin, indicating its universal conservation and functional importance related to replication origins. The stem-and-loop structure in S. gregaria/C. parallelus appears to be closely related to that found in Drosophila despite occupying a different position, and may be potentially associated with a second-strand-replication origin. This in turn suggests that such a secondary structure might be widely conserved across invertebrates while their location in the control region may be variable. We have looked for such a conserved structure in the control regions of two other insects, G. firmus and A. mellifera, whose DNA sequences have been published, and their possible presence is discussed.Mitochondrial control regions characterized to date in five different insect taxa (Drosophila, G. firmus, A. mellifera, S. gregaria, and C. parallelus) may be classed into two distinct groups having different evolutionary patterns. It is observed that tandem repetition of regions containing a probable replication origin occurred in some species from disjunct lineages in both groups, which would be the result of convergent evolution. We also discuss the possibility of a mechanism of parahomologous recombination by unequal crossing-over in mitochondria, which can explain the generation of such tandemly repeated sequences (especially the first critical repetition) in the control region of mtDNA, and also their convergent evolution in disjunct biological lineages during evolution. Correspondence to: G.M. Hewitt     

Adjustment of flight speed of gregarious desert locusts (Orthoptera: Acrididae) flying side by side

In tethered flying locusts, optomotor thrust responses induced by translatory pattern motion within the lateral visual fields were studied under closed-loop conditions. By modulating thrust in a compensatory manner, locusts counteracted a bias motion superposed on the thrust-related motion. This way, pattern speed was kept at 0° s^–1, indicating the set point of the respective optomotor control circuit. Though the quality of bias compensation varied greatly, it was largely independent from pattern characteristics. It might indicate that the gain of behavior not only is controlled by an automatic mechanism but also is affected by spontaneous modulations. Compensation of bias motion was critically dependent on the relation between self- and bias-generated motion: Locusts did not take control over pattern motion if self- and bias-generated motion differed greatly. Instead, locusts adopted a constant, supposingly preferred, thrust value. Therefore, flight speed is assumed to be controlled by two systems: the optomotor and a preferred thrust system. In free flight, an equalization of the flight speed of locusts within a swarm might result from similar behavior. In combination with a presumed coordination of the locusts' course direction, this may explain the continued cohesion of swarms in the field.     

GABA-like immunoreactivity in the suboesophageal ganglion of the locust Schistocerca gregaria

Summary Neurones in the suboesophageal ganglion of the locust Schistocerca gregaria were stained with an antiserum raised against gamma amino butyric acid (GABA). This ganglion consists of the fused mandibular, maxillary and labial neuromeres. Immunoreactive cell bodies of similar size and distribution occur in the lateral, ventral and middorsal regions of all three neuromeres. Approximately 200 cell bodies stain in both the mandibular and maxillary neuromeres and 270 in the labial neuromere. A few distinctly larger cells occur in the ventral groups and one large pair occurs in the lateral group of the maxillary neuromere. Dorsal commissures DCIV and DCV are composed mainly of stained fibres, while DCI–DCIII are largely unstained. A ventral commissure also stains in the maxillary neuromere. All longitudinal tracts contain both stained and unstained fibres. Many processes within the neuropil are also immunoreactive. A stained axon is found in the posterior tritocerebral commissure which enters the anterior dorsal region of the mandibular neuromere. The salivary branch of the 7th nerve contains one stained axon and two axons stain in nerve 8 which innervates neck muscles.     

Ultrastructure and immunocytochemistry of endocrine cells in the midgut of the desert locust,Schistocerca gregaria (Forskal)

Summary The endocrine cells of the midgut epithelium of the desert locust are found dispersed among the digestive cells and are similar to those of the vertebrate gut. According to their reactivity to silver impregnation techniques and the ultrastructural features of the secretory granules (shape, electron-density, size, and structure) 10 types of endocrine cell have been identified, of which seven are located in the main segment of the midgut or in the enteric caeca, and the other three seem to be present only in the ampullae through which the Malpighian tubules drain into the gut. The endocrine cells have a slender cytoplasmic process that reaches the gut lumen, a feature that supports the receptosecretory nature postulated for this cellular type in insects as well as vertebrates. Antisera directed against mammalian gastrin, CCK, insulin, pancreatic polypeptide and bombesin reacted with some of the endocrine cells. This is the first time that insulin- and bombesin-like immunoreactive cells have been described in the midgut of an insect.     

Purification and characterization of pyruvate carboxylase from the honeybee and some properties of related biotin-containing proteins in other insects

Our discovery of a M_r 128 k biotin-containing protein in an extract of the white-fly was an unexpected outcome of the use of an avidin-biotin-peroxidase method to visualize a western blot. This major biotin-containing protein was shown to be present in several tissues of 10 different species of insects by doing a western blot and staining it with streptavidin-linked peroxidase. The amount of this protein in the thorax of the mosquito, Aedes aegypti, increased during development. The non-flying grasshopper, Barytettix psolus, had reduced amounts of this protein in their thoraces compared to a flying grass-hopper, Schistocerca americana. The major biotin-containing protein was purified from the thoraces of honeybee, Apis mellifera, using an avidin Sepharose affinity column. The purified biotin-containing protein was shown to be pyruvate carboxylase, an enzyme that catalyzes the specific transfer of a carboxyl group to pyruvate, yielding oxaloacetate. The purified honeybee pyruvate carboxylase was characterized enzymatically and structurally. This protein had a single subunit of M_r 128 k and formed a native molecule of about M_r 500 k consisting of four of these subunits. The amino acid composition of the protein was also obtained. The enzymatic activity of this protein required acetyl-CoA, ATP, and Mg²⁺. The K_ms of the enzyme for bicarbonate and pyruvate were similar to pyruvate carboxylase from other oganisms. The biotin-containing protein was also partially purified from mosquito thoraces using the same methods and was shown to be pyruvate carboxylase. The comparison between insect pyruvate carboxylase and that of other organisms is provided and the possible physiological role of the pyruvate carboxylase in the thoracic muscles of insects is discussed.     

Quantitative studies on anti-allatin and lethal effects of a synthetic precocene in different larval instars of the desert locust

The lethal and anti-allatin effects of various doses of 7-ethoxy-precocene II (PIII), topically applied to crowdedSchistocerca gregaria nymphs, 0–27 h after the previous moult, were investigated for the five nymphal instars. The LD (lethal dose) probit lines were parallel for the different instars. The ED (effective anti-allatin dose) probit lines were also parallel. But for each instar the LD and ED probit lines seemed to be convergent toward the higher doses. The LD₅₀ and the ED₅₀ (g PIII/g fresh weight) were not widely separated; the LD₅₀/ED₅₀ ratio was instar dependent and varied between 1.26 (Vth instar) and 5.35 (IIIrd instar). Thus, separation between lethal effect, anti-allatin effect and no effect was rather poor; indeed in many instances all these types of results were obtained in a group of hoppers treated with the same dose in the same instar. A low precocene susceptibility was observed for the Ist instar and for each subsequent instar the susceptibility further decreased; the LD₅₀ and the ED₅₀ were respectively 886 g/g and 328 g/g for the Ist instar, but increased to 14 307 g/g and 11 367 g/g for the Vth instar. The anti-allatin effect of the precocene induced various morphogenetic responses: (1) permanent adultiform in the instar next to that treated; (2) death in the moult from the next to the second consecutive instar; (3) permanent adultiform in the second consecutive instar. The occurrence and/or distribution of these responses were instar dependent and for the first three instars also limitedly dose dependent. The anti-allatin effect of the precocene applied to Vth (last) instar nymphs became overt by sterility and lack of yellow colour in the adult. An overt anti-allatin effect was always coupled with a response dependent increase of the duration of the instar treated and/or of the next instar.

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Etudes quantitatives des effets létaux ou antiallates d'un précocène de synthèse sur différents stades larvaires du criquet pélerin

Résumé L'étude a porté sur les effets létaux ou antiallates de différentes doses de 7-éthoxy-précocène II (PIII), déposées de 0 à 27 heures après la mue sur la surface dorsale de l'abdomen de larves groupées de chacun des 5 stades larvaires deSchistocerca gregaria. Les courbes des doses létales (LD) ont été parallèles pour les différents stades. Les courbes pour les doses antiallates (ED) étaient elles aussi parallèles. Cependent pour chaque stade, ces deux courbes semblaient converger et même se croiser vers les doses les plus fortes. Les LD₅₀ et ED₅₀ (en g de PIII par gramme de poids frais) n'étaient pas nettement séparées; le rapport LD₅₀/ED₅₀ dépendait du stade et variait entre 1,26 (5è stade) et 5,35 (3è stade). Ainsi la distinction entre effet létal, effet antiallate et absence d'effet était plutôt faible; à la vérité, dans de nombreux cas, tous ces effets ont été obtenus dans un groupe de criquets traités avec la même dose au même stade. Une sensibilité assez faible au précocène a été observée, même au ler stade, et pendant chaque stade suivant la sensibilité ultérieure diminuait; LD₅₀ et ED₅₀ ont été respectivement 886 g/g et 328 g/g pour le ler stade, mais ont augmenté à 14 307 g/g et 11 367 g/g pour le 5è stade. Les effets antiallates ont induit différentes conséquences morphologiques: (1) des adultiformes permanents lors du stade suivant immédiatement le stade traité; (2) mort pendant la mue du ler ou 2nd stade suivant le traitement; (3) des adultiformes permanents dans le 2nd stade après le traitement. L'apparition et/ou la distribution de ces réponses dépendaient du stade, et pour les 3 premiers stades étaient partiellement dépendantes de la dose. L'effet antiallate du précocène au 5è stade (dernier) devenait évident avec la stérilité et la perte de la coloration jaune de l'adulte. Un effet antiallate manifeste était toujours associé à un accroissement d dépendant de la dose de la durée du stade traité et/ou du stade suivant.

     

Discharge patterns of locust visual interneurones

The spike discharges of the descending contralateral movement detector (DCMD) neurone, and of some smaller visual interneurones (S-units), were recorded in the ventral nerve cord of adult Schistocerca gregaria Forsk., in response to a stationary disc (25^o) or a small spot (0.2^o) stimulus. The discharge rate of each neurone was plotted over a period of 5 s exposure; the total number of spikes in this period was also noted. The DCMD response to the 25^o disc was a high-frequency burst falling off quickly to a low rate; the 0.2^o spot evoked a prolonged discharge with an early peak in rate. In S-units the discharge was prolonged with both targets; the rate rose to an early peak in each case, with a much higher rate for the larger disc. For the DCMD the total number of spikes per stimulus (5 s) was greater for the 0.2^o spot; for S-units it was greater for the 25^o disc. Thus an increase of about 30-fold in the number of ommatidia stimulated resulted in a fall in the total DCMD response to about one-quarter; a similar increase evoked a rise of about 6-fold in the S-unit response. When the 25^o disc was presented at progressively reduced intensities the total spike response of the DCMD rose steadily to a maximum at about 2.9 μ W m^-2; using the same procedure the spike output of S-units, initially high, declined monotonically. The role of inhibition in these results is discussed.