The dark-colour-inducing neurohormone of locusts in relation to an albino mutant of Schistocerca gregaria

In the albino mutant of an Okinawa strain of Locusta migratoria (L.) (Orthoptera: Acrididae), albinism is caused by the absence of the dark‐colour‐inducing neurohormone (DCIN), which is present in the corpora cardiaca (CC) of normally coloured phenotypes. This study tests whether the absence of DCIN is responsible for albinism in an albino mutant of another locust, Schistocerca gregaria (Forsk.) (Orthoptera: Acrididae). This seemed feasible because a single Mendelian unit controls albinism in both species. However, implantation of CC, or injection of an extract of CC, from albino donors of S. gregaria, induce dark coloration in crowded nymph recipients of the Okinawa albino mutant of L. migratoria, as effectively as do implanted CC, or injections of extract of CC, from normal phenotype donors of S. gregaria. Therefore, DCIN is present in the albino mutant of S. gregaria, and consequently, the albinism in this mutant is not caused by its absence. Implantation of CC, or injection of extracts of CC, from albino donors of S. gregaria to conspecific albino nymphs does not induce darkening. Only extremely high doses of synthetic DCIN injected into albino nymphs of S. gregaria are effective, inducing some darkening. The dose to induce such darkening in albino nymphs of S. gregaria is 50 nmol, ≈ 5 × 10⁶ times higher than that (10 femtomol) needed to induce equivalent darkening in nymphs of the Okinawa albinos of L. migratoria. The results are discussed and some possible explanations of the observed effects outlined.     

Maturation of locust corpora allata during the reproductive cycle: Effect of reserpine on allatotropic activity,juvenile hormone-III biosynthesis,and oocyte development

Reserpine, at doses of 20–175 μg per g body weight, severely retards oogenesis in newly emerged adult female migratory locusts (Locusta migratoria migratorioides) but does not increase mortality during the first 9 days and only slightly delays somatic growth. Total protein, and hemolymph vitellogenin content particularly, are significantly reduced in reserpine-treated locusts. The synthesis of juvenile hormone III (JH-III) following adult emergence, essential for induction of vitellogenesis and subsequent oogenesis, is dependent on the maturation and activation of the corpora allata (CA). CA of 7- to 8-day-old female locusts, treated with reserpine at day 1 after adult emergence, are only marginally active in vitro and are only slightly stimulated by an allatotropic factor. The basal activity and response of CA from the reserpine-treated locusts resembles that of newly emerged locusts, suggesting that reserpine specifically retards the initial maturation of the locust CA. Recovery of basal CA activity is evident on days 12–13 in reserpine-treated locusts, but responsiveness to the allatotropic factor is not recovered. Starvation of newly emerged females for 3 days and subsequent feeding did not effect ooctye development or CA activity. Cerebral content of the allatotropic factor, assayed on days 7–8, is not reduced by the reserpine treatment.     

Partial purification and characterization of locust allatotropin I

Methanol extracts of locust brains, corpora cardiaca (CC), and suboesophageal ganglia (SOG) were separated by gradient and/or isocratic reverse-phase high-performance liquid chromatography (HPLC) and allatotropic activity monitored in the eluted fractions. A major peak of activity, separated by isocratic separation with 12% 2-propanol, designated allatotropin I, exhibited identical retention times in the three tissue extracts. Doseresponse curves of allatotropin I indicate similar content in brain and CC-equivalents, whereas optic lobes, similarly separated by isocratic HPLC, contain only one-tenth of this amount of allatotropin. Allatotropin I is resistant to boiling and is susceptible to tryptic and chymotryptic digestion. Methanol extracts of thoracic muscle, Malpighian tubules, fat body or ovaries, similarly prepared and boiled, did not exhibit allatotropic activity at high doses of tissue equivalents.     

The morphology of suboesophageal ganglion cells innervating the nervus corporis cardiaci III of the locust

Summary The nervus corporis cardiaci III (NCC III) of the locust Locust migratoria was investigated with intracellular and extracellular cobalt staining techniques in order to elucidate the morphology of neurons within the suboesophageal ganglion, which send axons into this nerve. Six neurons have many features in common with the dorsal, unpaired, median (DUM) neurons of thoracic and abdominal ganglia. Three other cells have cell bodies contralateral to their axons (contralateral neuron 1–3; CN 1–3). Two of these neurons (CN2 and CN3) appear to degenerate after imaginal ecdysis. CN3 innervates pharyngeal dilator muscles via its anterior axon in the NCC III, and a neck muscle via an additional posterior axon within the intersegmental nerve between the suboesophageal and prothoracic ganglia. A large cell with a ventral posterior cell body is located close to the sagittal plane of the ganglion (ventral, posterior, median neuron; VPMN). Staining of the NCC III towards the periphery reveals that the branching pattern of this nerve is extremely variable. It innervates the retrocerebral glandular complex, the antennal heart and pharyngeal dilator muscles, and has a connection to the frontal ganglion.Abbreviations AH antennal heart - AN antennal nerves - AO aorta - AV antennal vessel - CA corpus allatum - CC corpus cardiacum - CN1, CN2, CN3 contralateral neuron 1–3 - DIT dorsal intermediate tract - DMT dorsal median tract - DUM dorsal, unpaired, median - FC frontal connective - FG frontal ganglion - HG hypocerebral ganglion - LDT lateral dorsal tract - LMN, LSN labral motor and sensory nerves - LN+FC common root of labral nerves and frontal connective - LO lateral ocellus - MDT median dorsal tract - MDVR ventral root of mandibular nerve - MVT median ventral tract - NCA I, II nervus corporis allati I, II - NCC I, II, III nervus corporis cardiaci I, III - NR nervus recurrens - NTD nervus tegumentarius dorsalis - N8 nerve 8 of SOG - OE oesophagus - OEN oesophageal nerve - PH pharynx - SOG suboesophageal ganglion - T tentorium - TVN tritocerebral ventral nerve - VLT ventral lateral tract - VIT ventral intermediate tract - VMT ventral median tract - VPMN ventral, posterior, median neuron - 1–7 peripheral nerves of the SOG - 36, 37, 40–45 pharyngeal dilator muscles     

Immunocytochemical localization of lipophorins in the flight muscles of the migratory locust (Locusts migratoria) at rest and during flight

Summary Locust lipoproteins (lipophorins) were localized by indirect immunofluorescence- and immunogold labelling in cryosections of dorsolongitudinal flight muscles. Immunolabelling was performed with monoclonal antibodies against apolipoprotein epitopes that are exposed at the surfaces of the lipophorin particles. Both at rest and during flight, lipophorins were located only in the wider spaces of the extracellular matrix, in the basement membranes of the individual muscle fibers and in the extracellular spaces that surround interfibrillar tracheoles. No internalization of lipophorins by the flight muscle cells was observed. Our results indicate that the unloading of lipophorins at the flight muscles is an extracellular event. Similarities with the vertebrate system of chylomicron and very-low-density lipoprotein degradation are discussed.     

Ultrastructural study of cholecystokinin-like immunoreactivity in endocrine cells of the insect midgut

Summary Electron-microscopic immunocytochemistry for the demonstration of CCK-like material in basal endocrine cells of the midgut of Aeschna cyanea, Locusta migratoria, Carausius morosus and Periplaneta americana was performed by use of the peroxidase-antiperoxidase procedure and the colloidal gold method. Immunoreactive cells appeared scattered among digestive and regenerative cells of the epithelium. Immunoreactivity was specifically detected over round to oval electron-dense granules whose size appeared rather different from species to species. Thus, the average size of 30% (d30) of the largest granules ranged from 312 nm in Periplaneta americana to 159 nm in Carausius morosus with intermediate values in Aeschna cyanea (d30=195 nm) and Locusta migratoria (d30=225 nm).     

Ageing adipokinetic cells in Locusta migratoria: an ultrastructural morphometric study

Summary A morphometric study was made of the ultrastructure of adipokinetic cells in resting adults of Locusta migratoria at 3, 23, and 43 days after imaginal ecdysis. The nucleus, rough endoplasmic reticulum, and Golgi apparatus enlarge with age, which indicates that the synthesis and packaging of secretory substances increases during ageing. The size of the storage compartment, consisting of secretory and ergastoplasmic granules, does not increase earlier than 23–43 days after imaginal ecdysis. The lysosomal compartment markedly enlarges between 3 and 23 days; later on, the growth of this compartment, especially of autophagosomes, is less prominent. This suggests that lysosomal destruction initially compensates for the production of new secretory granules, assuming that exocytosis of secretory granules by adipokinetic cells is insignificant in resting locusts. Afterwards, lysosomal destruction may no longer be sufficient to prevent over-production of secretory granules, as is suggested by the increase in the number of these granules between 23 and 43 days. This coincides with the appearance of a considerable number of large ergastoplasmic granules, which represent a spatially more efficient form of storage of secretory material than the much smaller secretory granules. The increase with age in the amount of secretory products indicates that the biosynthetic activity of the adipokinetic cells is not (finely) tuned to their releasing activity.     

Food selection by locusts: The rôle of learning in rejection behaviour

The food selection behaviour of male fifth instar nymphs of Locusta migratoria was monitored on the host plant wheat and on the non-host plants Senecio vulgaris, S. jacobaea and Brassica oleracea. The non-hosts were rejected, but the mode of rejection altered with time in a way which suggested associative learning. This hypothesis was tested and the results discussed in relation to classical theories of learning.

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La sélection des aliments chez les criquets: Le rôle de l'apprentissage dans le comportement de rejet

Résumé Des larves mâles de cinquième stade de Locusta migratoria ont été placées dans des cages avec des végétaux qui pouvaient être changées sans troubler les insectes. Le contact avec des pieds de blé entraînait généralement une palpation suivie de morsures et alors d'alimentation continue. Les plantes non-hôtes Senecio vulgaris, S. jacobaea et Brassica oleracea étaient rejetées, d'abord après morsures suivant la palpation, mais ultérieurement par palpation seule. Nous avons vérifié l'hypothèse que la sensation obtenue par palpation était initialement inadéquate pour provoquer le rejet, ce qui était assuré par les morsures ultérieures mais qu'avec l'expérience l'insecte apprenait à lier la sensation de palpation avec celle de morsures et progressivement rejetait après palpation seule. Ainsi les 8 premiers contacts avec des aliments désagréables ont été observés, soit quant S. vulgaris était présenté continuellement, soit quand S. vulgaris était remplacé par une autre plante désagréable après 4 contacts. Une analyse mathématique des résultats révèle une tendance au rejet par palpation seule à travers les contacts successifs, et que cette tendance est interrompue et se restaure quand l'espace végétal est changé. Ces résultats sond discutés dans le contexte des théories de l'apprentissage et on en a conclu que le phénomène observé est le résultat d'un apprentissage associatif.

     

The association of proctolin with a ventral abdominal muscle of Locusta migratoria

The association of proctolin with the external ventral protractor muscle of the VIIth abdominal segment (M234) of Locusta migratoria was investigated using immunohistochemistry and RP-HPLC in conjunction with the sensitive locust oviduct bioassay. Immunohistochemistry of whole-mount tissues revealed two proctolin-like immunoreactive axons in N2B_2b1 (the nerve branch which innervates M234) as well as immunoreactive processes and varicosities on the surface of M234. Immunogold staining of M234 demonstrated that the proctolin-like immunoreactivity was present in electron-dense granules in its motor terminals. A material indistinguishable from proctolin and with proctolin-like bioactivity co-eluted with authentic proctolin on two different RP-HPLC systems. Bath application of proctolin at concentrations greater than 10^-11 M resulted in a dose-dependent increase in the neurally-evoked fast twitch amplitude and duration of M234. Concentrations greater than 10^-9 M resulted in a dose-dependent increase in basal tonus of M234. These results indicate that proctolin, or a peptide very similar to proctolin, is present in the motor innervation of M234 and acts as a cotransmitter and/or neuromodulator at this typical fast skeletal muscle.Abbreviations M234 external ventral protractor muscle of the Vllth abdominal segment - RP-HPLC reverse-phase high pressure liquid chromatography - TFA trifluoroacetic acid     

Separating food and water deprivation in locusts: effects on the patterns of consumption, locomotion and growth

Abstract. In a factorial experiment, fifth-instar Locusta migratoria (L.) (Orthoptera: Acrididae) were given either dry food (lyophilized grass) and drinking water, food only, water only, or neither food nor water.Food consumption and insect weight were measured daily, and the behaviour of each locust was recorded for 5 h on each of four consecutive days and for 2.5 h on the fifth.Consumption declined progressively in locusts given food only, and those given water only were not observed to drink after the first day of food deprivation.The decline in food consumption on the first day was accounted for by a decrease in the average duration of feeds, which remained constant thereafter.The further decline in consumption over subsequent days was due to a progressive decline in the number of feeds.Although food availability did not slow weight loss relative to locusts given neither food nor water, the availability of water without food did.The proportion of time locomoting increased in all deprivation treatments, but the pattern of change across the five observation days differed markedly between treatments.Locusts given food but no water increased locomotion from 20% of the time budget (the value for controls) to 30% on the first day of deprivation, and by the second day had reached a plateau of approximately 65%.which was maintained until the experiment was terminated on day 5.In contrast, locusts given water but no food approached the 65% level of locomotion on the first day, which was stutistically greater than the 55% observed in those deprived of both food and water.This increase was due both to an increase in the number of locomotion bouts initiated and an increase in the average duration of locomotion bouts.On the second and third days, all deprivation treatments maintained locomotion at around 65%.By day 4, locomotion had decreased to approximately 15% in locusts deprived of both food and water, but not in those deprived of food only or water only.Unlike those given only food, locusts given only water showed a reduction in locomotion of c. 15% on the fifth day.