Spatial distribution of synapses onto thoracic motor neurones in locusts

Summary Aggregates of synaptic vesicles, stained black by the zinc iodide-osmium procedure, can be visualised with the light microscope in 1 m plastic sections. This allows the main branches of a neurone to be reconstructed relatively rapidly and the associated vesicle aggregates to be plotted. By resectioning, the identity of the vesicle aggregates has been confirmed with the electron microscope. Two flight motor neurones in the mesothoracic ganglion of the locust have been examined. One is identified as a dorsal longitudinal muscle motor neurone (muscle 112) and the other is probably a subalar neurone (muscle 99). Both have a large density of vesicle aggregates on the neuropilar segment, the widest part of the main neuronal axis, but few on the neurite within 250 m of the cell body. The larger branches arising from the neuropilar segment tend to have a lower density of aggregates than fine branches, which suggests that synapses to the branches may occur mainly on the distal twigs. These results are an important preliminary step in determining the integrative functions of such neurones and have immediate implications in the interpretation of microelectrode recordings.JSA is supported by grant KU 240/3 from the Deutsche Forschungsgemeinschaft to Dr. W. Kutsch. We thank Mrs. Christine Davies for valuable assistance with the resectioning technique.     

A broadly tuned odorant receptor in neurons of trichoid sensilla in locust,Locusta migratoria

Insects have evolved sophisticated olfactory reception systems to sense exogenous chemical signals. Odorant receptors (ORs) on the membrane of chemosensory neurons are believed to be key molecules in sensing exogenous chemical cues. ORs in different species of insects are diverse and should tune a species to its own specific semiochemicals relevant to their survival. The orthopteran insect, locust (Locusta migratoria), is a model hemimetabolous insect. There is very limited knowledge on the functions of locust ORs although many locust OR genes have been identified in genomic sequencing experiments. In this paper, a locust OR, LmigOR3 was localized to neurons housed in trichoid sensilla by in situ hybridization. LmigOR3 was expressed as a transgene in Drosophila trichoid olfactory neurons (aT1) lacking the endogenous receptor Or67d and the olfactory tuning curve and dose-response curves were established for this locust receptor. The results show that LmigOR3 sensitizes neurons to ketones, esters and heterocyclic compounds, indicating that LmigOR3 is a broadly tuned receptor. LmigOR3 is the first odorant receptor from Orthoptera that has been functionally analyzed in the Drosophila aT1 system. This work demonstrates the utility of the Drosophila aT1 system for functional analysis of locust odorant receptors and suggests that LmigOR3 may be involved in detecting food odorants, or perhaps locust body volatiles that may help us to develop new control methods for locusts.     

The modulation of skeletal muscle contraction by FMRFamide-related peptides of the locust

The external ventral protractor muscle of the VIIth abdominal segment, M234, is a skeletal muscle that possesses receptors that recognize a range of FMRFamide-related peptides and application of these peptides results in an increase in the amplitude of neurally evoked contractions with little or no effect on basal tonus. FLRFamide itself has the same biologic activity as the extended peptides, whereas truncation to LRFamide or RFamide results in a peptide with no biologic activity. The receptors recognizing these extended FLRFamides, which include SchistoFLRFamide, seem to be different from the SchistoFLRFamide receptors found on locust oviduct visceral muscle. SchistoFLRFamide and the non-peptide mimetic, benzethonium chloride (Bztc), increase the frequency and amplitude of miniature endplate potentials, increase the amplitude of neurally evoked excitatory junction potentials, and result in a hyperpolarisation of resting membrane potential. Bztc, however, also abolishes the active membrane response that may explain its ability to decrease neurally evoked contractions.     

Origin and development of unusual insect muscle tension receptors

This work describes the origin and development of about 200 tension receptor cells located around the anterior attachment site of the locust ovipositor muscle and their migration to their final position on the muscle fibres. The locust ovipositor muscle is the only insect system in which more than 100 tension receptor cells are associated with a single muscle. Neuronal precursors of tension receptors are first detectable by horseradish peroxidase immunohistochemistry in fourth instar larvae. Precursors consist of cell clusters (doublets, triplets and quadruplets) located on the anterior attachment site of the muscle. In the early fifth larval stage, cell clusters are absent, although a few sensory neurons that lie embedded between the muscle fibres are apparent. These neurons send their dendrites towards the anterior end of the muscle fibres and their axons posteriorly. By the fourth day of the fifth larval stage, a large number of cell clusters appears on the anterior muscle attachment site. In addition to these assemblies, cells have been identified that extend long processes running exactly along the lateral margin of the attachment site. These cells are thought to provide navigating cues for migrating tension receptors, since they are absent in later stages. By the end of the fifth larval stage, most of the clusters gradually disappear and increasing numbers of differentiated neurons embedded between the muscle fibres become visible. We conclude that the majority of tension receptors develop during the last larval stage from precursors situated on the muscle apodeme. They then migrate from the apodeme to their final place on the muscle fibres where they assume an appropriate orientation.     

Production of α-galactosidase by a novel actinomycete <Emphasis Type="Italic">Streptomyces griseoloalbus</Emphasis> and its application in soymilk hydrolysis

α-Galactosidase production by a newly isolated actinomycete Streptomyces griseoloalbus under submerged fermentation was investigated. The influence of initial pH of medium, incubation temperature, inoculum age and inoculum size on α-galactosidase formation was studied. Various carbon sources were supplemented in the medium to study their effect on enzyme production. The influence of the concentration of locust bean gum on enzyme production also was optimized. Optimization of process parameters resulted in a highest α-galactosidase activity of 20.4 U/ml. The highest α-galactosidase activity was obtained when the fermentation medium with initial pH 6.0 and containing 1% locust bean gum as growth substrate was inoculated with 10% (v/v) of 72 h grown inoculum and incubated at 30°C. The hydrolysis of flatulence-causing oligosaccharides in soymilk by the enzyme was also investigated. Thin layer chromatographic analysis of enzyme-treated soymilk samples showed the complete hydrolysis of soy oligosaccharides liberating galactose, the final product.     

蝗虫减数分裂的实验

目的：探讨经COL处理后蝗虫减数分裂的动态变化和温湿度的环境变化与细胞分裂的关系。方法：①分别在蝗虫腹腔注入不同浓度(1．2mg／kg,2．4mg／kg,3．6mg／kg,4．8mg/kg)的COL,5h后制片观察减数分裂各时相的细胞动态变化。②记录研究一年中5-11月份温湿度变化与体重、细胞分裂的相关关系。结果：①经COL处理的各剂量组中,蚯虫减数分裂粗线期后各时相细胞数目比对照组均有不同程度的差异(P＜0．01),其中以4．8mg／kg剂量的处理组效果较理想。②蝗虫体重低于0．5g时,细胞分裂数较少。当体重在0．5g以上时,减数分裂各时期和有丝分裂期细胞数均有明显的增加。③温度增高,蝗虫细胞分裂数增加,且以7月和11月份细胞分裂指数为最多。结论：蝗虫经COL处理后减数分裂各分裂时期细胞数有明显增加;当蝗虫体重增加,环境温度升高,细胞分裂各个时期的细胞数目也增多。     

A Novel Spore Wall Protein from Antonospora locustae (Microsporidia: Nosematidae) Contributes to Sporulation

Microsporidia are obligate intracellular parasites, existing in a wide variety of animal hosts. Here, we reported AlocSWP2, a novel protein identified from the spore wall of Antonospora locustae (formerly, Nosema locustae, and synonym, Paranosema locustae), containing four cysteines that are conserved among the homologues of several Microspodian pathogens in insects and mammals. AlocSWP2 was detected in the wall of mature spores via indirect immunofluorescence assay. In addition, immunocytochemistry localization experiments showed that the protein was observed in the wall of sporoblasts, sporonts, and meronts during sporulation within the host body, also in the wall of mature spores. AlocSWP2 was not detected in the fat body of infected locust until the 9th day after inoculating spores via RT‐PCR experiments. Furthermore, the survival percentage of infected locusts injected with dsRNA of AlocSWP2 on the 15th, 16th, and 17th days after inoculation with microsporidian were significantly higher than those of infected locusts without dsRNA treatment. Conversely, the amount of spores in locusts infected with A. locustae after treated with RNAi AlocSWP2 was significantly lower than those of infected locusts without RNAi of this gene. This novel spore wall protein from A. locustae may be involved in sporulation, thus contributing to host mortality.     

Mechanosensitive potassium channels in locust muscle membrane

Patch clamp recordings have been made from adult locust (Schistocerca gregaria) muscle membrane to study the mechanosensitivity of potassium channels (BK and IK) in cell-attached patches by transiently applying measured pressures to the contents of the patch pipettes. The aim of the investigations was to demonstrate a novel gating behaviour by pressure of the BK channel in contrast to the familiar behaviour of the IK channel. The open probability (p ₀) of the IK channel increased rapidly in response to a pressure step and monotonically during a pressure ramp. This gating was readily repeatable and rapidly reversible. The relationship between ln[p ₀/(1–p ₀)] and transmembrane pressure was linear. In comparison, p ₀ for the BK channel was also increased by pressure, but its gating was delayed, cumulative, and hysteretic. Received: 12 July 1998 / Revised version: 7 October 1998 / Accepted: 7 October 1998     

Morphology of the central projections of physiologically characterised neurones from the locust metathoracic femoral chordotonal organ

Summary The metathoracic femoral chordotonal organ of the locust (Locusta migratoria) is an internal proprioceptor composed of mechanosensory neurones which respond to tibial position, velocity, or acceleration, or to combinations of these parameters. Discriminant function analyses confirmed the visual observation that neurones with different responses to tibial movements had different central branching patterns. Some aspects of the projections were consistent for all neurones (e.g., the path taken by the main neurite through the metathoracic ganglion), whereas other regions of branches were consistently reduced or missing in some response classes. Some position-and-acceleration receptors had no main branches off the main neurite, and must therefore make relatively restricted contact with motor neurones and interneurones. Phasic or tonic neurones which responded in ranges of tibial extension had branches which projected further medial in Dorsal Commissures III and IV than similar neurones which responded in ranges of tibial flexion. I compare my results to previous studies of mapping in the insect CNS.Abbreviations (ms) (mt)FCO (mesothoracic) (metathoracic) femoral chordotonal organ - ANOVA Analysis of Variance     

Oxygen consumption of flying locusts

ABSTRACT. Desert locusts ( Schistocerca gregaria Forskal.) were flown into an airstream from a wind tunnel while tethered to a balance. Rate of oxygen consumption and lift were recorded. The mean rate of oxygen consumption of male locusts in level flight at airspeeds between 2.5 and 3m/s was found to be 12.6ml/g/h.