蝗虫染色体分带技术的研究

本文对采自长沙岳麓山的中华稻蝗〔Oxya chinensis (Thunberg)〕和采自杭州的小稻蝗〔Oxya hyla intricata (Stal) 〕进行了染色体C带、R带、N带和A带的处理研究。结果表明,C带显示结构异染色质,其位置在不同分裂相中较为稳定;R带即G带的反带,带纹较C带丰富,在粗线期最为明显,本文观察到不同的温育条件可影响R带产生的效果;N带和Ag带可相互参照比较,从而为NOR的准确定位提供依据;A带即荧光带,本文对其显带程序作了一定的改进。 Abstract:In this paperOxya chinensis (Thunberg) collected from the Yuelu Mountains in Changsha, Hunan Province andOxya hyla intricata (Stal) collected from Hangzhou, Zhejiang province were analysed by chromosomal C-banding、R-banding、N-banding、Ag-banding and A-banding. The results showed that the constitutive heterochromatin was stained in C-banding, and it relatively stable; R-banding was the reverse of G-banding, its bands were relatively obvious at the pachytene and abundent, it may be because of the selective extracts that cause the bands evident, the different effects were surveyed under the various incubation conditions; Both N-banding and Ag-banding can stain the nucleolar organizer and the two were compared in this paper; The A-banding was a little improved in the light of the banding process.     

Further Characterization of the Antidiuretic Factor From the Glandular Part of the Corpora Cardiaca of the Migratory Locust

Summary

Two antidiuretic hormones are present in the corpora cardiaca of the locust, Locusta migratoria. They are located in the nervous and the glandular parts (NCC and GCC) respectively. A procedure was developed to avoid contamination of GCC by NCC. The peptide nature of the GCC hormone was established and its structural stability was studied.

Additional evidence regarding the role of cyclic AMP as second messenger of the GCC antidiuretic factor was presented.     

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.     

Nitric oxide: a co-modulator of efferent peptidergic neurosecretory cells including a unique octopaminergic neurone innervating locust heart

Our findings suggest that nitric oxide (NO) acts as peripheral neuromodulator in locusts, in which it is commonly co-localized with RF-like peptide in neurosecretory cells. We also present the first evidence for NO as a cardio-regulator in insects. Putative NO-producing neurones were detected in locust pre-genital free abdominal ganglia by NADPH-diaphorase histochemistry and with an antibody against NO synthase (NOS). With both methods, we identified the same 14 somata in each examined ganglion: two dorsal posterior midline somata; six ventral posterior midline somata; and three pairs of lateral somata. A combination of NOS-detection methods with nerve tracing and transmitter immunocytochemistry revealed that at least 12 of these cells were efferent, of which four were identified as peptidergic neurosecretory cells with an antiserum detecting RFamide-like peptides. One of the latter was unequivocally identified as an octopaminergic dorsal unpaired median (DUM) neurone, which specifically projected to the heart (“DUM-heart”). Its peripheral projections revealed by axon tracing appeared as a meshwork of varicose endings encapsulating the heart. NOS-like immunoreactive profiles were found in the heart nerve. NO donors caused a dose-dependent increase in heart rate. This cardio-excitatory effect was negatively correlated to resting heart rate and seemed to be dependent on the physiological state of the animal. Hence, NO released from neurones such as the rhythmically active DUM-heart might exert continuous control over the heart. Possible mechanisms for the actions of NO on the heart and interactions with other neuromodulators co-localized in the DUM-heart neurone (octopamine, taurine, RF-amide-like peptide) are discussed.P.A.S. is the leading author, following the tragic death of Alexander Bullerjahn on 20.12.03.This work was supported by the Deutsche Forschungsgemeinschaft (grant number SFB 515; project of H.-J.P.).     

Temperature-sensitive gating in a descending visual interneuron, DCMD

Activity in neural circuits can be modified through experience-dependent mechanisms. The effects of high temperature on a locust visual interneuron (the descending contralateral movement detector, DCMD) have previously been shown to be mitigated by prior exposure to sub-lethal, elevated temperatures (heat shock, HS). Activity in the DCMD is reduced at high temperature in naïve animals (control), whereas HS animals show a maintained spike count at all temperatures. We examined whether this finding was due to direct effects of temperature on visual processing, or whether other indirect feedback mechanisms were responsible for the observed effect in the DCMD. Activity in the DCMD was elicited using a computer-generated looming image, and the response was recorded extracellularly. The temperature of visual processing circuits contributes directly to HS-induced plasticity in the DCMD, as maintaining the brain at 25°C during a thoracic temperature ramp eliminated the high frequency activity associated with HS. Removing ascending input by severing the thoracic nerve cord reduced DCMD thermosensitivity, indicating that indirect feedback mechanisms are also involved in controlling the DCMD response to increased thoracic temperature. Understanding how thermosensitive feedback within the locust affects DCMD function provides insight into critical regulatory mechanisms underlying visually-guided behaviors.     

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。     

Neuronal connections between central and enteric nervous system in the locust, Locusta migratoria

The number and location of neurons, in the central nervous system, that project into the frontal connective was studied in the locust by using retrograde neurobiotin staining. Staining one frontal connective revealed some 70 neurons in the brain. Most of these were located within both tritocerebral lobes. Additional groups of neurons were located within the deutocerebrum and protocerebrum. Some 60 neurons were labelled in the suboesophageal ganglion. These formed nine discernable populations. In addition, two neurons were located in the prothoracic ganglion and two neurons in the first abdominal neuromere of the metathoracic ganglion. Thus, some 250 neurons located within the head ganglia, and even neurons in thoracic ganglia, project into the ganglia of the enteric nervous system. This indicates that the coordination between the central and enteric ganglia is much more complex than previously thought. With the exception of some previously described dorsal unpaired median neurons and a few motor neurons in the head ganglia, the identity and function of most of these neurons is as yet unknown. Possible functions of the neurons in the thoracic ganglia are discussed.     

Specific activity of a Bacillus thuringiensis strain against Locusta migratoria manilensis

Bacillus thuringiensis (Bt) has played an important role in biocontrol of pests. However, insecticidal activity of B. thuringiensis against locusts has been rarely reported. Bt strain BTH-13 exhibiting specific activity to locusts was isolated from a soil sample in China and characterized. Its bipyramidal parasporal crystal is mainly composed of a protein of 129 kDa, and produces a mature toxin of 64 kDa after activation. The pattern of total DNA from BTH-13 showed a large and three small plasmid bands. Known δ-endotoxin genes, cry1Aa, cry1Ab, cry1Ac, cry1C, cry3, cry4 and cry7Aa were not found from strain BTH-13 by PCR amplification. The sequence analysis of a DNA fragment produced by PCR amplification with degenerate cry-selective primers revealed that the fragment encoded a δ-endotoxin segment, which exhibited some similarity to several Cry proteins (41% of the highest similarity to Cry7Ba1). Toxicity tests were performed against Locusta migratoria manilensis, and the results demonstrated that trypsin-treated sporulated cultures and crystal proteins had high toxicity to larval and adult locusts. Cry toxin of BTH-13 was detected on the midguts of treated locusts using immunofluorescent technology, which confirmed the site of action of the crystal proteins in their toxicity for locusts.     

Structural implications on the interaction of scorpion alpha-like toxins with the sodium channel receptor site inferred from toxin iodination and pH-dependent binding

The alpha-like toxin from the venom of the scorpion Leiurus quinquestriatus hebraeus (Lqh III) binds with high affinity to receptor site 3 on insect sodium channels but does not bind to rat brain synaptosomes. The binding affinity of Lqh III to cockroach neuronal membranes was fivefold higher at pH 6.5 than at pH 7.5. This correlated with an increase in the electropositive charge on the toxin surface resulting from protonation of its four histidines. Radioiodination of Tyr(14) of Lqh III abolished its binding to locust but not cockroach sodium channels, whereas the noniodinated toxin bound equally well to both neuronal preparations. Radioiodination of Tyr(10) or Tyr(21) of the structurally similar alpha-toxin from L. quinquestriatus hebraeus (LqhalphaIT), as well as their substitution by phenylalanine, had only minor effects on binding to cockroach neuronal membranes. However, substitution of Tyr(21), but not Tyr(14), by leucine decreased the binding affinity of LqhalphaIT approximately 87-fold. Thus, Tyr(14) is involved in the bioactivity of Lqh III to locust receptor site 3 and is not crucial for the binding of LqhalphaIT to this site. In turn, the aromatic ring of Tyr(21) takes part in the bioactivity of LqhalphaIT to insects. These results highlight subtle architectural variations between locust and cockroach receptor site 3, in addition to previous results demonstrating the competence of Lqh III to differentiate between insect and mammalian sodium channel subtypes.     

Response decrement in an auditory neurone of the locust

An identified interneurone (SA3) of the suboesophageal ganglion has an axon in both circumoesophageal connectives ascending to the brain. This is a novel morphology for a cell in the auditory system of the locust. The neurone is also novel for its physiological responses to sound in that it displays side-dependent response decrement (habituation). Responses to a tone directed at one ear decrement without affecting responses from the other ear. The responses on the decremented side recover when the opposite ear is stimulated. The decrement is the result of a diminishing amplitude of the compound excitatory post-synaptic potential but no inhibition is seen. Response decrement does not occur if the stimulus frequency is varied.