菊方翅网蝽内生殖系统的结构与形态发育研究

一般认为网蝽科及其近缘类群所特有的伪储精囊与其它半翅目异翅亚目昆虫的储精囊具有相同的储存精子的功能,但近期的功能形态学研究否定了伪储精囊的储精功能并认定其为雌性生殖附腺。本文从成虫性成熟过程中的内生殖系统发育角度,描述了菊方翅网蝽Corythucha marmorata的卵巢、侧输卵管、伪储精囊、精巢、精囊和雄性生殖附腺的结构及其形态变化。在成虫性成熟过程中,雄虫内生殖器官精囊和雄性附腺逐渐加长,且成熟期的雄性附腺充满粉红色分泌物;雌虫内生殖器官的成熟过程分为卵巢小管的卵室形成、卵黄沉积和卵粒成熟三个阶段,且排完卵后卵巢重新孕卵,出现周期性形态变化;交配时侧输卵管基部膨大为精液接受器,并接受精液(含精子和精浆);交配1d后侧输卵管恢复为正常状态,精液或至少部分精浆弥漫性渗入伪储精囊,并在伪储精囊内形成黄棕色沉积核;未交配雌虫的伪储精囊一直保持透明状,而已交配雌虫的伪储精囊具有明显的黄棕色沉积核。据精液传递和粉红色雄性生殖附腺分泌物渗入雌虫伪储精囊两个关键证据推断,菊方翅网蝽雌虫的伪储精囊具有储存精液的功能。     

昆虫雄性附腺功能蛋白研究进展

昆虫雄性附腺蛋白是精液蛋白的主要来源,对雌雄虫生殖过程具有重要生理功能,按功能可分为精包结构蛋白和功能蛋白两类。精包结构蛋白参与精包的形成;功能蛋白在交配过程中随精子一起转移到雌虫体内,导致雌虫行为和生理的深刻变化,如降低雌虫再交配率、提高产卵量、促进精子转移、储存和竞争等。随着对昆虫雄性附腺功能蛋白研究的深入,特别对果蝇附腺功能蛋白的详细研究,从分子水平上阐述蛋白质序列与功能的关系,明确其作用机制,可为进一步阐明昆虫生殖和进化机制等提供新依据。     

蜱雄性附腺分泌物及其功能的研究进展

交配是蜱类繁殖过程的关键环节,诱发雌蜱发生一系列的生理变化,并最终产卵。蜱雄性附腺分泌物在交配过程中发挥着重要作用,具有保护、活化精子,促进受精、卵巢发育和卵黄发生的功能,并对雌蜱的生殖生理行为等产生影响,如诱导雌蜱快速吸血和加速产卵。本文在简要分析蜱雄性附腺结构和分泌物生化特性基础上,系统阐述了蜱雄性附腺分泌物中各种功能因子的研究现状,着重论述其在精子获能、诱导雌蜱吸血、促进雌蜱卵巢发育和卵黄发生等方面的进展,并对未来研究提出了展望,以期为此领域的研究拓展思路。     

桑天牛雄性附腺内容物组分分析

测定桑天牛Apriona germaric(Hope)雄性附腺内容物中各组分含量及变化情况。结果表明,内容物中可溶性蛋白占总腺管鲜重的8.39%;总糖占总鲜重的4.21%;海藻糖和游离氨基酸分别占总鲜重的0.50%、0.25%。随虫龄的增大,内容物中各组分含量不断降低。交配过程中,雄性附腺内容物部分转移到雌虫,交配后附腺内容物中总糖、海藻糖、游离氨基酸等组分含量均降低,蛋白质组分含量先升高再降低,48h即恢复可到交配前水平。大、小附腺内各组分含量变化有差异。     

昆虫雄性生殖腺分泌物的功能

昆虫两性成功交配后,雄性生殖腺分泌物使雌性昆虫的生理和行为都发生了巨大的改变。昆虫雄性生殖腺分泌物含有多种具有生物活性的分子,这些生物活性分子通过成功交配转移到雌虫生殖道后,对雌虫的生殖活动产生影响,使交配雌虫一段时间内不再交配,使已转移的精子易于在雌虫生殖道内储藏,使卵与精子完成受精过程,还可刺激雌虫产卵和卵的发育,调控排卵和产卵等生殖过程。在精子的转移过程中,雄性生殖腺分泌物中的抗菌媒介质能使雌虫的生殖导管提供友好的环境。此外,一些昆虫的雄性生殖腺分泌物还含有一些有毒的化学物质,保护已产下的卵不被天敌取食和病原体侵染。     

中华真地鳖的生殖系统

通过解剖分析中华真地鳖EupolyphagasinensisWalker雌雄内生殖系统和外生殖器的构造,比较不同生长期雌虫的内生殖系统变化,并依据雌虫外生殖器和最后一腹节的构造叙述了地鳖虫卵鞘的形成过程。结果表明:中华真地鳖雌性附腺和雄性附腺发达;雄性外生殖器的阳体结构较为复杂;雌虫外生殖器、腹末构造与卵鞘表面形态的形成有必然的关系。     

亚洲柑橘木虱内生殖系统形态变化规律研究

亚洲柑橘木虱Diaphorina citri Kuwayama是柑橘黄龙病的传播媒介。本文利用Ste REO Discovery V20体视显微镜对亚洲柑橘木虱成虫内生殖系统进行解剖观察,并探索了亚洲柑橘木虱雌雄成虫内生殖系统的形态变化规律。结果表明:雄虫内生殖系统由1对精巢、1对输精管、1个精泵、1个射精管、1对附腺和1个贮精囊组成。雌虫内生殖系统由1对卵巢、1对侧输卵管、1个中输卵管、2个附腺、1个黏腺和1个受精囊组成。交配前期和交配期的雄虫精巢饱满,精巢在交配后期明显萎缩。交配期和交配后期的雄虫贮精囊都明显大于交配前期的贮精囊。雌虫受精囊在交配前期、交配期和交配后期依次增大,交配前期的受精囊呈不饱满状态,交配期和交配后期受精囊呈饱满状态,内有白色精包。交配后期的雌虫卵巢内有大量成熟的橙黄色卵。     

桑天牛成虫交配后血淋巴和生殖系统内可溶性总糖和蛋白质含量的变化

以生理状况相似的未交配桑天牛Apriona germari Hope雌、雄成虫为供试昆虫, 观察、记录交配活动;采集血淋巴,解剖生殖系统并绘图;用蒽酮比色法和福林-酚法检测交配前、后成虫血淋巴和生殖系统内可溶性总糖和蛋白质含量变化。结果表明：交配后1 h,桑天牛雄虫血淋巴内的可溶性总糖含量增加21.38%,蛋白质含量降低22.66%;雄虫生殖系统内的可溶性总糖和蛋白质含量都明显升高;雄性附腺作为某些特异性蛋白质的合成场所其内的可溶性总糖和蛋白质含量分别降低81.76%和63.76%,雌虫血淋巴和卵巢内的可溶性总糖和蛋白质含量都升高。交配后, 雄虫发生陪伴行为最短历时为4 h, 可能是其重要的生殖策略之一。     

斑蜡蝉生殖系统的解剖与组织学

详细描述了斑蜡蝉两性生殖系统各部分的结构与组织学。精巢由6条互相分离的精巢管组成,而卵巢则含有14条典型的端滋式卵巢管。用组织化学方法测定了雄性附腺各段的分泌物的成分,它们分别构成精包、精液与精包鞘。订正了过去文献中关于雌性生殖系统某些部分的解释:过去误称为雌性附腺的其实是受精囊及其腺,而过去误称为受精囊的其实是接纳及消化精包的交配囊。受精囊腺的细胞具有一条曲折的细胞内分泌小管,贯穿内膜开口于腺腔。交配囊体及交配囊管上皮折突的内层为腺细胞,有孔道通过内膜与腔相通,它可以分泌一种(或多种)酶以消化精包。此外还从卵黄发生的速率与精包授精的关系,讨论了精包作为一种蛋白养料的来源,在雌虫营养与生殖上的重要性。     

柑橘大实蝇的内生殖系统形态结构

柑橘大实蝇Bactrocera minax(Enderlein),是柑橘的重要害虫。本文基于光学显微镜、扫描电镜、石蜡切片观察,对柑橘大实蝇的内生殖系统形态结构进行研究。结果表明,柑橘大实蝇雌虫内生殖系统主要由卵巢、侧输卵管、中输卵管、受精囊、附腺、前生殖腔(包含布氏交配囊、桑葚腺、生殖腔片)、后生殖腔(阴道和产卵针)组成。雄虫内生殖系统主要由精巢、精泵、输精管、附腺、输入射精管、输出射精管、后附腺和阳茎组成。其雌虫有泄殖腔,位于产卵针前端稍后,雄虫无泄殖腔。雌虫前生殖腔表面被2对肌肉包裹,内部的布氏交配囊、桑葚腺和生殖腔片不易被观察。精泵是一个淡黄色球体,由射精突(精泵内骨骼)、肌纤维(肌肉)、射精囊组成。柑橘大实蝇的内生殖形系统形态结构或组织经过进化,从而适应其伪产卵器的运动、交配、产卵等行为机制。为理解昆虫繁殖生理、进化和多样性,以及昆虫的产卵、交配和代谢物排泄等行为机制提供理论基础。     

The activity of platelet activating factor-acetyl hydrolase (PAF-AH) in the salivary glands of Rhodnius prolixus

In this work, we investigated the activity of the platelet activating factor acetyl hydrolase (PAF-AH) in the salivary gland homogenates and saliva of Rhodnius prolixus. PAF-AH activity in the salivary gland homogenates was lower than in the saliva. Preliminary characterization of the enzyme demonstrated that it hydrolyzed the substrate 2-thio-PAF, was detectable just in 1 pair of salivary gland homogenates in 0.5 ml buffer, and was stable under different conditions. PMSF, TPCK, TLCK, pepstatin A and p-BPB all inhibited the PAF-AH activity. Enzyme specific activity in salivary gland homogenates diminished immediately after feeding of 5th-instar larvae, and increased before feeding by adult insects. 2-Thio-PAF induced platelet-aggregation that was inhibited by previous incubation of the substrate with salivary gland homogenates or saliva. The relevance of PAF-AH for providing Rhodnius with a feeding mechanism for facilitating the sucking of a high volume of blood meal in a short period is discussed.     

Esterases as pesticide biomarkers in crayfish (Procambarus clarkii, Crustacea): tissue distribution, sensitivity to model compounds and recovery from inactivation

The specific activities of acetyl- and butyrylcholinesterase and carboxylesterase were assayed in the digestive gland and in nervous and muscle tissues of the crayfish Procambarus clarkii. Since acetylcholinesterase prevails in nervous tissue and carboxylesterase in digestive gland, they are proposed as biomarkers. Muscle had negligible activities of all esterases, and all tissues had a low butyrylcholinesterase activity. Esterases were mostly cytosolic in digestive gland and muscle, but membrane-bound in nervous tissue; use of Triton X-100 is not recommended due to its widely diverging effects in esterase assays. Phenylmethylsulphonylfluoride inhibited acetyl- and butyrylcholinesterase in extracts from all tissues, and in digestive gland only carboxylesterase. In digestive gland, tetra[monoisopropyl]-pyrophosphorotetramide inhibited all esterases with different sensitivities, while in muscle and nervous tissue it only partially inhibited all esterases. Carbamates inhibited 100-fold more strongly than organophosphates acetyl- and butyrylcholinesterase activities. Carboxylesterase was inhibited by carbaryl and chlorpyrifos, but not by eserine and malathion. In vitro conditions to evaluate recovery from inactivation of esterases by model pesticides were established for acetylcholinesterase and carboxylesterase. The new reactivation protocol could be useful as a biomarker of pesticide exposure to differentiate between dilution-reversible inhibitions, indicating carbamate exposure, from dilution-irreversible effect, attributed to organophosphate exposure.     

A novel RGD-toxin protein,Lj-RGD3, from the buccal gland secretion of Lampetra japonica impacts diverse biological activities

RGD (Arg-Gly-Asp) motif toxin proteins from snake venoms, saliva glands secretion of leech or tick have typical characteristics of inhibiting platelet aggregation, angiogenesis, and tumor growth. Here we report cloning and characterization of a novel RGD-toxin protein from the buccal gland of Lampetra japonica. In an attempt to study the activities of anticoagulant in the buccal gland secretion of L. japonica, we established buccal gland cDNA library and identified a gene encoding a predicted protein of 118 amino acids with 3 RGD motifs. The predicted protein was named Lj-RGD3. We generated the cDNA of Lj-RGD3 and obtained the recombinant protein rLj-RGD3. The polyclonal antibodies against rLj-RGD3 recognized the native Lj-RGD3 protein in buccal gland secretion in Western blot analyses. The biological function studies reveal that rLj-RGD3 inhibited human platelet aggregation in a dose-dependent manner with IC₅₀ value at 5.277 μM. In addition, rLj-RGD3 repressed bFGF-induced angiogenesis in the chick chorioallantoic membrane model. rLj-RGD3 also inhibited the adhesion of ECV304 cells to vitronectin. Furthermore, rLj-RGD3 induced apoptosis and significantly inhibited proliferation, migration, and invasion evoked by bFGF in ECV304 cells. Taken together, these results suggested that rLj-RGD3 is a novel RGD-toxin protein possessing typical functions of the RGD-toxin protein.     

Cell surface expression of 4 beta-galactosyltransferase accompanies rat parotid gland hypertrophy induced by changes in diet.

Maintenance of rats for 2 weeks on a diet consisting of 50% inert cellulose and 50% laboratory chow resulted in hypertrophy of the parotid gland and a 4-fold increase in total membrane-associated 4 beta-galactosyltransferase enzyme activity (EC 2.4.1.38). Localization of the increased specific activity to the cell surface of the enlarged gland was shown by subcellular fractionation of Golgi and plasma membranes. This observation was confirmed by enzyme assays of intact cells; quantification of immunofluorescence was made by using a fluorescence activated cell sorter. Parotid gland hypertrophy was inhibited by the administration of the specific modifier protein alpha-lactalbumin as well as by a monospecific antibody for 4 beta-galactosyltransferase. These agents also inhibited the incorporation of thymidine into DNA.     

Effects of wheat germ agglutinin and concanavalin A on parameters of highly purified sodium-potassium adenosine triphosphatases from Squalus acanthias and Electrophorus electricus.

The effects of two lectins, wheat germ agglutinin and concanavalin A, were studied on a variety of parameters of two highly purified (Na+ + K+)-ATPases (ATP phosphohydrolase, EC 3.6.1.3), from the rectal salt gland of Squalus acanthias and from the electroplax of Electrophorus electricus. Both lectins agglutinated the rectal gland enzyme equally, but wheat germ agglutinin inhibited (Na+ + K+)-ATPase activity much more. The electroplax enzyme was only marginally agglutinated and inhibited by the lectins. Neuraminidase treatment of the rectal gland (Na+ + K+)-ATPase had no effect on germ agglutinin inhibition. The inhibition of the rectal gland (Na+ + K+)-ATPase by wheat germ agglutinin could be reversed by N,N'-diacetylchitobiose, which has a high affinity for wheat germ agglutinin. Neither ouabain inhibition nor ouabain binding to the rectal gland enzyme was affected by wheat germ agglutinin. The p-nitrophenylphosphatase activity of the rectal gland enzyme was not inhibited by wheat germ agglutinin. Na+-ATPase activity, which reflects ATP binding and phosphorylation at the substrate site was inhibited by wheat germ agglutinin and this inhibition was reversed by potassium. Evidence is cited (Pennington, J. and Hokin, L.E., in preparation) that the inhibition of the (Na+ + K+)-ATPase by wheat germ agglutinin is due to binding to the glycoprotein subunit.     

A study of l-leucine, l-phenylalanine and l-alanine transport in the perfused rat mammary gland: possible involvement of LAT1 and LAT2

The transport of l-leucine, l-phenylalanine and l-alanine by the perfused lactating rat mammary gland has been examined using a rapid, paired-tracer dilution technique. The clearances of all three amino acids by the mammary gland consisted of a rising phase followed by a rapid fall-off, respectively, reflecting influx and efflux of the radiotracers. The peak clearance of l-leucine was inhibited by BCH (65%) and d-leucine (58%) but not by l-proline. The inhibition of l-leucine clearance by BCH and d-leucine was not additive. l-leucine inhibited the peak clearance of radiolabelled l-leucine by 78%. BCH also inhibited the peak clearance of l-phenylalanine (66%) and l-alanine (33%) by the perfused mammary gland. Lactating rat mammary tissue was found to express both LAT1 and LAT2 mRNA. The results suggest that system L is situated in the basolateral aspect of the lactating rat mammary epithelium and thus probably plays a central role in neutral amino acid uptake from blood. The finding that l-alanine uptake by the gland was inhibited by BCH suggests that LAT2 may make a significant contribution to neutral amino acid uptake by the mammary epithelium.     

Identification of anticoagulant activities in the salivary glands of the soft tick,Ornithodoros savignyi

Salivary gland extracts of the sand tampan, Ornithodoros savignyi, prolonged the activated partial thromboplastin time (APTT) and prothrombin time (PT) significantly in a concentration-dependent manner. There was also a pronounced inhibition of human activated factor Xa (fXa) by salivary gland extracts. The salivary gland extracts inhibited chromogenic assays specific for both fXa and thrombin. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) of the salivary gland proteins followed by elution of specific areas or bands from a polyvinylidene difluoride (PVDF)-membrane, showed that various anticoagulant factors are present when screened by means of the APTT assay. The most active component was associated with a band of M _r of 14 kDa. Partial purification of this component was achieved using isoelectric focusing (IEF) and size-exclusion highperformance liquid chromatography (HPLC).     

The mysid Siriella armata,a biological model for the study of hormonal control of molt and reproduction in crustaceans: a review

Summary

The mysid Siriella armata provides a new biological model for investigations on the molting and reproductive physiology in crustaceans. The main endocrine centres (Y-organ, mandibular organ, androgenic gland, X-organ and sinus gland) have been described and are available for experimentation. Experimental cautery of Medulla Interna-Medulla Externa-X-organ-sinus gland complex (MI-ME-X-organ-SG) of the eyestalk inhibited molt and brood production demonstrating that the complex plays a role in regulation, at least via a positive action upon the circulating ecdysteroids. In the present paper, the results already obtained are reviewed and the perspectives offered by this biological model discussed in reference to other crustaceans.     

Rapid inhibition of lipogenesis in vivo in lactating rat mammary gland by medium- or long-chain triacylglycerols and partial reversal by insulin.

An intragastric load of medium- or long-chain triacylglycerols inhibited lipogenesis in lactating rat mammary gland in vivo by 82 or 89% respectively. This inhibition was reversed partially by insulin administration. Long-chain triacylglycerols inhibited hepatic lipogenesis in vivo but medium-chain triacylglycerols increased it 2-fold. Glucose utilization in vitro by mammary gland acini from triacylglycerol-fed rat was normal.     

Subacute physiological stress induced by phosphamidon on carbohydrate metabolism in midgut gland of prawn, Penaeus indicus

Midgut gland carbohydrate metabolism of penaeid prawn, Penaeus indicus was studied on acute and acclimation to sublethal concentration of phosphamidon. The midgut gland tissue of acclimated prawn showed inhibited glycolysis with an onset of gluconeogenesis. In general acclimation to sublethal concentration resulted in the elevation of the synthetic phase of midgut gland carbohydrate metabolism.