黄脸油葫芦消化道和马氏管的组织学观察

应用石蜡切片技术对黄脸油葫芦Teleogryllus emma(Ohmachi and Matsumura)成虫消化道和马氏管的显微结构进行观察。消化道由前肠、中肠和后肠3部分组成:前肠由内向外可分为6层:内膜、肠壁细胞层、底膜、纵肌、环肌和围膜;中肠组织结构也分为6层,即由内向外依次为围食膜、肠壁细胞层、底膜、环肌、纵肌和围膜;后肠的组织结构与前肠基本相似,但内膜比前肠的薄,且肌肉的排列较前肠不规则,与中肠的肌肉排列相似,即环肌在内,纵肌在外。消化道各部位的结构差异与功能有密切关系。马氏管管壁由8个左右形状多变并具有显著细胞核的大形的单层上皮细胞组成。     

桃小食心虫幼虫的消化道和马氏管的显微结构观察

利用光学显微镜和扫描电子显微镜,在形态学和组织学水平上研究_『桃小食心虫 Carposina sasakii 幼虫消化道和屿氏管的结构.桃小食心虫幼虫消化道由前肠、中肠和后肠组成.前肠细短,肌肉层薄.前肠与中肠交界处有突出的胃盲囊.中肠长且粗大,内有围食膜,肠壁细胞较大,外层为发达的环肌和纵肌.后肠上皮细胞内陷很深.6根念珠状的马氏管位于中、后肠分界处.     

庭疾灶螽中肠及马氏管结构

【目的】本研究旨在以庭疾灶螽Tachycines asynamorus为例探索驼螽消化系统和排泄系统在结构上与其生活环境的适应关系。【方法】运用解剖学方法、石蜡切片技术、冰冻切片技术及超薄切片技术对庭疾灶螽中肠及马氏管的结构进行研究。【结果】庭疾灶螽中肠向前延伸出3个胃盲囊包围着前胃。中肠上皮由再生细胞、柱状上皮细胞和内分泌细胞构成,具有典型的再生细胞龛;闭合型内分泌细胞紧贴在再生细胞龛的外围,基底区聚集大量的分泌颗粒。柱状上皮细胞内聚集有2类大的分泌颗粒：线团状颗粒和电子密度很高的球状颗粒;中肠管腔内有明显的围食膜结构,中肠基底部由基膜和肌肉层组成。马氏管着生在中后肠的交界处,从横切面看马氏管管壁具有3~5个细胞,细胞近管腔端部具有大量长微绒毛,细胞质内分布着电子致密的同心圆球晶体,基底膜内折形成膜迷路。【结论】庭疾灶螽中肠柱状上皮细胞的线团状颗粒由微丝包裹;内分泌细胞由再生细胞龛中的细胞分化而来,产生内分泌颗粒并将其排到血腔;中肠基膜发达,包含微丝与复合糖成分,基膜通过对中肠上皮细胞的支撑作用为肠道蠕动提供保障。庭疾灶螽马氏管细胞中可见大量颗粒和大量同心圆球晶体,推测可能是一种储存排泄。     

小地老虎变态期间马氏管超微结构与酯酶活性的变化

本实验用光镜和电镜观察了小地老虎Agrotis ypsilon Rottemberg幼虫在变态期间马氏管超微结构的变化及成虫马氏管的重组过程,同时还研究了变态期马氏管酯酶的活性.结果表明:(1)变态期间马氏管外形完整,除至预蛹期隐肾复合体解体外,其余无明显变化.(2)变态期间管壁细胞变化显著.幼虫6龄末期马氏管细胞结构开始变化,主要特点为:细胞质电子密度高,充满了核糖体颗粒,微绒毛萎缩,线粒体从萎缩的微绒毛中退出进入细胞质,基膜内褶破坏.进入预蛹期幼虫马氏管细胞解体:基膜内褶、顶端微绒毛、线粒体及细胞质内的其它细胞器消失,并形成自体吞噬泡,细胞质内仅存细胞核及各种类型的液泡.但是在变态期间因底膜始终存在,故马氏管外形不变;至蛹后期,成虫马氏管细胞在原位重组,基膜内褶由浅变深,微绒毛由短变长,线粒体内嵴从无到有.(3)变态过程中羧酸酯酶和酸性磷酸酯酶的活性变化趋势基本相同,以六龄幼虫最强,预蛹期次之,蛹期最低.     

小地老虎变态期间马氏管超微结构与酯酶活性的变化

本实验用光镜和电镜观察了小地老虎Agrotis ypsilon Rottemberg幼虫在变态期间马氏管超微结构的变化及成虫马氏管的重组过程,同时还研究了变态期马氏管酯酶的活性.结果表明:(1)变态期间马氏管外形完整,除至预蛹期隐肾复合体解体外,其余无明显变化.(2)变态期间管壁细胞变化显著.幼虫6龄末期马氏管细胞结构开始变化,主要特点为:细胞质电子密度高,充满了核糖体颗粒,微绒毛萎缩,线粒体从萎缩的微绒毛中退出进入细胞质,基膜内褶破坏.进入预蛹期幼虫马氏管细胞解体:基膜内褶、顶端微绒毛、线粒体及细胞质内的其它细胞器消失,并形成自体吞噬泡,细胞质内仅存细胞核及各种类型的液泡.但是在变态期间因底膜始终存在,故马氏管外形不变;至蛹后期,成虫马氏管细胞在原位重组,基膜内褶由浅变深,微绒毛由短变长,线粒体内嵴从无到有.(3)变态过程中羧酸酯酶和酸性磷酸酯酶的活性变化趋势基本相同,以六龄幼虫最强,预蛹期次之,蛹期最低.     

短额负蝗精子发生过程中SBA受体的动态分布

用常规组织学方法观察短额负蝗Atractomorpha sinensis Bolivar精子发生过程中生精细胞的显微结构,并以大豆凝集素(soybean agglutinin,SBA)为探针利用凝集素细胞化学方法研究该过程中N-乙酰半乳糖复合物的分布变化。结果表明,短额负蝗精子发生经历了精原细胞增殖期、初级精母细胞期、次级精母细胞期、精子细胞形成期和精子成熟期5个时期,在这5个时期中各期生精细胞的大小、形态、核染色体等变化明显。在整个精子发生过程中,N-乙酰半乳糖复合物出现于精原细胞期,并于精母细胞期发生明显的修饰和变化,精子形成期和成熟期没有N-乙酰半乳糖复合物的表达。提示,N-乙酰半乳糖复合物的修饰和变化与短额负蝗生精细胞的生长和分化密切相关。     

小地老虎马氏管细微结构的特点

本文通过光镜和电镜观察,研究了小地老虎grois ypsilon Rottemberg六龄幼虫和成虫马氏管及管壁细胞的形态特点和排泄方式.幼虫马氏管中不同细胞的分泌方式和亚细胞结构有很多差异,端段和中段的马氏管细胞基内褶发达,并发现在隐肾内的端段细胞中,有一类含有大量的线粒体.在幼虫中,胞吐排泄占有重要地位,并观察到有微绒毛顶部胞吐、微绒毛间胞吐和顶膜胞吐三类.成虫马氏管细胞主要有二种类型,即大型的基本细胞和小型的底细胞,前者为主,后者数量较少.基本细胞中存在复杂的液胞系,排泄以排放液胞为主.     

山西省六个负蝗种群的遗传多样性

用等位酶电泳分析方法对短额负蝗(A tractom orpha sinensis)和奇异负蝗(A tractom orpha p ereg rina)各3个自然种群10种酶(AAT,CK,G 3PD,HEX,IDH,LDH,M DH,M E,PG I,PGM)进行检测。结果显示:两种负蝗在某些基因座上共享常见的等位基因,如A a t-1-b、A a t-2-b、G 3p d-a、Ck-1-b和Ldh-b;除个别基因座在部分种群符合H ardy-W e inberg平衡外,两种负蝗的大多数基因座的基因型频率显著偏离H ardy-W e inberg平衡。此外,奇异负蝗M e-c(0.318~0.740)、短额负蝗H ex-1-a(0.800~1.000)及Ldh-b(0.487~0.750)等位基因频率呈现出由北向南递增的趋势,表明M e和H ex、Ldh基因座上的等位基因频率与地理分布存在一定的相关关系。短额负蝗平均每个基因座的等位基因数(A)为1.9~2.3、多态基因座百分率(P)为56.3%~68.8%、平均观察杂合度(Ho)为0.072~0.096;而奇异负蝗的相应值依次为A=1.7~2.2,P=43.8~56.3%,Ho=0.070~0.107。从A、P和Ho3个参数可知,短额负蝗遗传多样性明显高于奇异负蝗。6个负蝗种群的平均观察杂合度均明显低于H ardy-W e inberg平衡预期值,表明6个负蝗种群均出现了杂合体缺乏现象。短额负蝗3种群I值为0.971~0.996,奇异负蝗3种群I值为0.982~0.995,短额负蝗与奇异负蝗I值为0.379~0.451,表明种内遗传相似度明显高于种间,从种间I值可知奇异负蝗和短额负蝗属于近缘种。根据R oger's遗传距离进行的聚类分析表明,两种负蝗可分为两支,且两种负蝗的遗传距离与地理距离均存在明显的相关趋势。两种负蝗的平均FST值都不显著偏离0值(奇异负蝗FST=0.087,p>0.05,短额负蝗FST=0.045,p>0.05),表明该两种负蝗种群间的分化不明显。     

东方粉蝶幼虫马氏管的超微构造分化

东方粉蝶幼虫有6条马氏管,每条管可以简易地分为四个部分:直肠导,迴肠纲,黄色段和白色段,所有这四个部分首要细胞的顶面都折叠形成微绒毛(特别是迴肠纲,其顶面进一步折叠形成微道),线粒体几乎延伸到微绒毛顶端。在基部,大量基膜折叠形成细胞内管道,向细胞顶部延伸。在细胞质中,有许多线粒体,糙面内质纲和空泡。每一个细胞的细胞核中有一些散的染色质物质,并且外形呈不规则形。在黄色段首要细胞中,有大量证明是由矿物质沉积形成的电子密集颗粒,及猜疑为微孢子原虫的细胞质囊球。上述每段可能具有的功能会在这报告中讨论。     

短额负蝗配子发生过程中蓖麻凝集素受体的分布

利用凝集素细胞化学方法对短额负蝗Atractomorpha sinensis Bolivar配子发生过程中蓖麻凝集素（Ricinus communis agglutinin-Ⅰ,RCA-Ⅰ）受体分布进行了定位研究,旨在认识昆虫配子发生过程中β-半乳糖复合物的分布规律及意义。结果表明： 在短额负蝗精子发生过程中,RCA-Ⅰ在各期生精细胞膜上均有明显的阳性反应,变形期及成熟期精子头部为中强度阳性反应。卵子发生中,各阶段卵母细胞均为阳性反应,在卵母细胞生长期阳性反应达峰值,随着卵黄颗粒的逐渐形成,阳性反应逐渐减弱,成熟卵子中卵黄膜上重新出现中强度阳性反应。结果提示短额负蝗配子发生过程中β-半乳糖复合物的高度表达和修饰变化与其配子发生密切相关。     

The use of eryopreservation for convenient bioassay of insect organs in vitro

Abstract Pheromone glands of the female moth Helicoverpa (Heliothis) armigera and Malpighian tubules of the locust Locusta migratoria migratorioides (R&F) retained partial biological activity after eryopreservation. Pheromone glands were frozen to -70^oC in a tissue culture medium (TC-199) containing 10% of the cryoprotectant dimethylsulfoxide. Malpighian tubules, however, required initial preincubation for 24 h in a medium of high osmolarity before being frozen to -70^oC in the presence of 10% of the cryoprotectant glycerol. After rapid thawing of both organs, biological activity was compared with the respective fresh organs. Using a radiochemical assay, it was shown that previously frozen pheromone glands were stimulated by pheromone biosynthesis activating neuropeptide to the same degree as fresh pheromone glands. In addition, previously frozen Malpighian tubules were stimulated by an extract of corpora cardiaca to produce c-AMP to the same extent as fresh Malpighian tubules.     

白蜡虫马氏管的超微结构

白蜡虫Ericerus Pela的马氏管由两条黄色膨泡串状的端管和一条公共管构成,通过公共管与消化道相连。端管和公共管细胞结构相似,都具有非胶原质的基膜,高度发达的基褶, 长而致密的微绒毛,微绒毛无线粒体插入,细胞质中线粒体少,且随机分布。细胞质的绝大部分为两种矿质-尿酸颗粒结晶所占据,一种为不规则结晶,另一种为轮纹状结晶。白蜡虫马氏管可能发生了合胞化,其排泄方式可能是一种以滞留排泄为主,离子梯度排泄方式为辅的特有的排泄方式。     

Calcium transport across the basolateral membrane of isolated Malpighian tubules: a survey of several insect orders

The Malpighian tubules play a major role in haemolymph calcium homeostasis in insects by sequestering excess Ca²⁺ within the biomineralized granules that often accumulate in the tubule cells and/or lumen. Using the scanning ion‐selective microelectrode technique, measurements of basolateral Ca²⁺ transport are determined at several sites along the length of the Malpighian tubules isolated from the eight insects representing seven orders: Drosophila melanogaster (Diptera), Aedes aegypti (Diptera), Tenebrio molitor (Coleoptera), Acheta domesticus (Orthoptera), Trichoplusia ni (Lepidoptera), Periplaneta americana (Blattodea), Halyomorpha halys (Hemiptera) and Pogonomyrmex occidentalis (Hymenoptera). Ca²⁺ transport is specific to tubule segments containing Ca‐rich granules in D. melanogaster and A. aegypti, whereas Ca²⁺ transport is relatively uniform along the length of whole tubules in the remaining species. Generally, manipulation of second messenger pathways using cAMP and thapsigargin has little effect on rates of basolateral Ca²⁺ transport, suggesting that previous effects observed across midtubules of A. domesticus are unique to this species. In addition, the present study is the first to provide measurements of basolateral Ca²⁺ across single principal and secondary tubule cells, where Ca²⁺ uptake occurs only across principal cells. Estimated times for all tubules to eliminate the entire haemolymph Ca²⁺ content in each insect range from 6 min (D. melanogaster) to 19 h (H. halys) or more, indicating that rates of Ca²⁺ uptake by the Malpighian tubules are not always rapid. The results of the present study suggest that the principal cells of the Malpighian tubules contribute to haemolymph calcium homeostasis by sequestering excess Ca²⁺, often within specific tubule segments.     

Intracellular melanization of the larvae of Dirofilaria immitis in the malpighian tubules of the mosquito, Aedes sollicitans

Frequent melanization of larvae of the nematode Dirofilaria immitis parasitizing the Malpighian tubules of the mosquito, Aedes sollicitans, has been observed. Melanized and nonmelanized larvae in the Malpighian tubules were examined using light and electron microscopy. The results indicate that the pattern of melanin deposition and the ultrastructural characteristics of the pigment around the worms are identical to that observed on nematodes which have undergone humoral melanization in other dipteran insects. In the Malpighian tubules, no contact between the intracellular melanized nematodes and the hemolymph or hemocytes was observed. The results suggest that the Malpighian tubules of this species of mosquito are capable of inducing a melanotic response to invading nematode parasites. It is proposed that this is an example of “humoral” melanization at an intracellular site.     

Proteomic changes in response to crystal formation in Drosophila Malpighian tubules

Kidney stone disease is a major health burden with a complex and poorly understood pathophysiology. Drosophila Malpighian tubules have been shown to resemble human renal tubules in their physiological function. Herein, we have used Drosophila as a model to study the proteomic response to crystal formation induced by dietary manipulation in Malpighian tubules. Wild-type male flies were reared in parallel groups on standard medium supplemented with lithogenic agents: control, Sodium Oxalate (NaOx) and Ethylene Glycol (EG). Malpighian tubules were dissected after 2 weeks to visualize crystals with polarized light microscopy. The parallel group was dissected for protein extraction. A new method of Gel Assisted Sample Preparation (GASP) was used for protein extraction. Differentially abundant proteins (p<0.05) were identified by label-free quantitative proteomic analysis in flies fed with NaOx and EG diet compared with control. Their molecular functions were further screened for transmembrane ion transporter, calcium or zinc ion binder. Among these, 11 candidate proteins were shortlisted in NaOx diet and 16 proteins in EG diet. We concluded that GASP is a proteomic sample preparation method that can be applied to individual Drosophila Malpighian tubules. Our results may further increase the understanding of the pathophysiology of human kidney stone disease.     

Effect of Na+, K+, ouabain,amiloride and ethacrynic acid on the transepithelial potential across Malpighian tubules of Locusta

A transepithelial potential of +8.74±0.29 mV (n = 85) has been recorded across the Malpighian tubules of Locusta. The effect of varying the Na⁺ and K⁺ concentration in the bathing medium on the transepithelial potential has been determined. The data show that the transepithelial potential does not obey the Nernst equation for K⁺. Ouabain, ethacrynic acid and amiloride all inhibit the transepithelial potential. The results are discussed in relation to the nature of the mechanisms of cation transport across the Malpighian tubules.     

蝉次目昆虫“足-消化道-马氏管”的形态/功能分化及其在蝉次目昆虫系统演化中的意义

对蝉次目3个总科(蝉总科,沫蝉总科,角蝉总科)代表类群(12种成虫)的足、消化道和马氏管的形态结构及其分化进行了比较形态学研究,提出蝉次目昆虫消化道及马氏管整体结构可被划分为两种类型的观点(蝉总科和沫蝉总科消化道及马氏管整体结构为同一种类型,角蝉总科消化道及马氏管整体结构为另一种类型),并分别绘制了两种类型的模式结构图;基于成虫的马氏管比较形态学研究,提出蝉次目昆虫成虫马氏管可被划分为6部分的观点,并分别绘制了角蝉科、叶蝉科、蝉科和沫蝉科成虫的马氏管区域分化模式结构图。研究结果表明,蝉次目昆虫的足、消化道、马氏管的形态、功能分化与其生物学、行为生态学特性等密切相关,其形态学特征既为蝉次目各类群的单系性提供了重要支持证据,也为进一步从行为学、生态学、生物学等方面探讨蝉次目的系统演化历史提供了新的信息;研究结果支持蝉总科和沫蝉总科为姊妹群的观点,即蝉次目3个总科的系统关系为角蝉总科+(蝉总科+沫蝉总科)。     

The effect of infection with Dirofilaria immitis (dog heartworm) on fluid secretion rates in the Malpighian tubules of the mosquitoes Aedes taeniorhynchus and Anopheles quadrimaculatus

Dirofilaria immitis, the parasitic nematode causing the disease dog heartworm, is transmitted by female mosquitoes. During their development, larval nematodes reside in the cells of the Malpighian tubules of these mosquitoes for approx 13 days. We have examined the effect of the presence of these large intracellular parasites on the main physiological function of the Malpighian tubules, i.e. fluid secretion. Rates of fluid secretion were examined in vitro using both normal and infected tubules of the mosquito species Aedes taeniorhynchus and Anopheles quadrimaculatus. Tubules of A. quadrimaculatus show changes in trasport rate during the reproductive cycle. Those of A. taeniorhynchus do not. Infection with larvae of D. immitis had no effect on the rate of fluid secretion in tubules of A. quadrimaculatus. In A. taeniorhynchus by contrast, the tubules show decline in transport with time following infection. The reduction in transport capacity is proportional to the number of worms infecting the tubule. The present paper and separate ultrastructural studies demonstrate that parallel changes in microvillar ultrastructure and epithelial transport rates occur in response to infection by the parasite. In both species examined, the survival of the mosquitoes and their vector potential are determined by factors other than the transport capacities of the infected Malpighian tubules.     

Transepithelial transport of salicylate by the Malpighian tubules of insects from different orders

The organic anion salicylate is a plant secondary metabolite that protects plants against phytophagous insects. In this study, a combination of salicylate-selective microelectrodes and a radioisotope tracer technique was used to study the transepithelial transport of salicylate by the Malpighian tubules of 10 species of insects from five orders. Our results show that salicylate is transported into the lumen of the Malpighian tubules in all the species evaluated, except Rhodnius prolixus. The transepithelial transport of salicylate by the Malpighian tubules of Drosophila simulans, Drosophila erecta, Drosophila sechellia, and Acheta domesticus was saturable, Na⁺-dependent and inhibited by α-cyano-4-hydroxycinnamic acid. This transport system resembles that previously found in tubules of Drosophila melanogaster. In contrast, transepithelial transport of salicylate by Malpighian tubules of Tenebrio molitor, Plagiodera versicolora, Aedes aegypti, and Trichoplusia ni was unaffected by Na⁺-free bathing saline. The presence of both salicylate and salicylate metabolites in the secreted fluid samples from the Malpighian tubules of A. domesticus, R. prolixus, T. molitor, and T. ni indicates that insect Malpighian tubules may both transport and metabolize salicylate. The highest capacities to rid the hemolymph of salicylate were found in T. molitor, P. versicolora and Drosphila spp. Our results suggest that transport of salicylate by the Malpighian tubules might contribute to elimination of this organic anion from the hemolymph, particularly in some species that encounter high levels of organic anion in the diet.