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.     

A structural basis for fluid secretion by malpighian tubules

The Malpighian tubules of Calliphora are described, emphasizing the possible role of surface specializations in solute-linked water transport. The tubules are composed of two cell types, primary and stellate, intermingling along the tubule length. The primary cells have long narrow basal infoldings and a microvillate luminal border, both intimately associated with mitochondria. The stellate cells have shorter and wider basal infoldings and their apical microvilli do not contain mitochondria. Application of the standing gradient hypothesis to this sytem provides a model for urine formation in which the local gradients for osmotic water flow occur within the long narrow channels of the basal infolds and microvilli of the primary cells. Stellate cells may modify the initial secretion by reabsorbing sodium.     

Regional specialization in the malpighian tubules of the new zealand glow-worm Arachnocampa luminosa (diptera: Mycetophilidae). The structure and function of type I and II cells

The Malpighian tubules of the glow-worm Arachnocampa luminosa are divided into four morphologically distinct regions (Parts 1--4) each comprised of a different cell type (Types I--IV). The ultrastructure of Type II cells is indicative of a transport function. The basal cell surface is highly invaginated and at the apical surface the lumen is lined with microvilli about 80% of which contain mitochondria. Spherites contained in these cells are formed from small vesicles produced by the Golgi apparatus. They have a central uric acid core enclosed by laminations of phosphates of calcium and magnesium. Cells of Part 2 of the tubule secrete a fluid high in potassium (173 mM) and low in sodium (18 mM). The cell is 30 mV negative and the lumen 44 mV positive to the bathing solution. This is consistent with the proposal of an apical cation pump. The secretion produced by Part 2 of the tubules is modified by the Type I cells by the reabsorption of potassium (162 mM) and the addition of sodium (24 mM) to the primary excretory fluid. Type I cells are 20 mV negative and the lumen 22 mV positive with respect to the bathing medium. From ultrastructural observations, Type I cells exhibit features characteristic of transporting cells thought to have an absorptive function. The basal and apical cell surfaces are extensively folded, and mitochondria are found in bands above the basal infoldings and below the microvilli. Mitochondria do not penetrate the microvilli. On comparative grounds, the fine structure of Type I cells suggest that they reabsorb ions from the tubule lumen. Energy for these processes may come from the breakdown of lipids by microperoxisomes contained within these cells. Alternatively, the fluid produced by Part 2 of the tubule may be modified passively by diffusional processes across Type I cells.     

Electron-microscopic study of the excretory system of hungry females of the tick Hyalomma asiaticum P. Sch. et E. Schl. 2]

In H. asiaticum the cells of the Malpighian tubules and these of the rectal cas have the uniform structure: the apical surface is covered with microvilli, the basal plasmatic membrane forms relatively small invaginations. As to ultrastructural characters, there is no distinct division of the Malpighian tubule into departments. The distal ends of the tubules are not only somewhat enlarged and form the so-called ampulla cells of which are noticeably flattened. The microvilli and basal folds of the plasmatic membrane in this area of the tubule are indistinct. The cells of the ampulla and the neighbouring area of the tubule are characterized by the presence of inclusions with mucopolysaccharide secretion confined by the membrane. The microvilli are most developed on cells of the proximal ends of the Malpighian tubules. Well developed microvilli of the rectal sac form a striated border each containing a microtube inside. The basal invaginations are developed here better than in the cells of the Malpighian tubules.     

Post-embryonic development of the Malpighian tubules in <Emphasis Type="Italic">Apis mellifera</Emphasis> (Hymenoptera) workers: morphology,remodeling, apoptosis,and cell proliferation

The honeybee Apis mellifera has ecological and economic importance; however, it experiences a population decline, perhaps due to exposure to toxic compounds, which are excreted by Malpighian tubules. During metamorphosis of A. mellifera, the Malpighian tubules degenerate and are formed de novo. The objective of this work was to verify the cellular events of the Malpighian tubule renewal in the metamorphosis, which are the gradual steps of cell remodeling, determining different cell types and their roles in the excretory activity in A. mellifera. Immunofluorescence and ultrastructural analyses showed that the cells of the larval Malpighian tubules degenerate by apoptosis and autophagy, and the new Malpighian tubules are formed by cell proliferation. The ultrastructure of the cells in the Malpighian tubules suggest that cellular remodeling only occurs from dark-brown-eyed pupae, indicating the onset of excretion activity in pupal Malpighian tubules. In adult forager workers, two cell types occur in the Malpighian tubules, one with ultrastructural features (abundance of mitochondria, vacuoles, microvilli, and narrow basal labyrinth) for primary urine production and another cell type with dilated basal labyrinth, long microvilli, and absence of spherocrystals, which suggest a role in primary urine re-absorpotion. This study suggests that during the metamorphosis, Malpighian tubules are non-functional until the light-brown-eyed pupae, indicating that A. mellifera may be more vulnerable to toxic compounds at early pupal stages. In addition, cell ultrastructure suggests that the Malpighian tubules may be functional from dark-brown-eyed pupae and acquire greater complexity in the forager worker bee.     

Ultrastructure of the malpighian tubules of Schistocerca gregaria

The ultrastructure of the Malpighian tubules of the adult desert locust, Schistocerca gregaria, is described. Male and female adults possess about 233 tubules, which empty proximally into the midgut-ileal region of the alimentary canal by way of 12 ampullae. The tubules vary from 10 mm to 23 mm in length. About one third of them are directed anteriorly, attaching distally at the caeca, while the remainder are directed posteriorly, attaching to other tubules, the rectum or large tracheal trunks adjacent to the hindgut. The Malpighian tubules from all locations examined consist of three ultrastructurally distinct regions: proximal, middle, and distal, referring to their position relative to the midgut. All cell types possess ultrastructural features characteristic of ion transporting tissue, i.e., elaboration of the basal and apical membranes and a close association of these membranes with mitochondria. The distal and proximal segments are short (1.5-1.7 mm) and heavily tracheated, and each is composed of a single, distinct cell type. The middle region is the longest segment of the Malpighian tubule and is composed of two distinct cell types, primary and secondary. Both cell types are binucleate. The more numerous primary cells have large nuclei, contain laminate concretions in membrane-bound vacuoles, and possess large microvilli that contain mitochondria. The secondary cells are smaller and possess smaller nuclei. The microvilli are reduced and lack mitochondria. Secondary cells do not contain laminate concretions. The possible compartmentalization of ion and fluid transport function based on segmentation in the Malpighian tubules is discussed.     

Micromorphology of the malpighian tubules in the louse Pediculus humanus corporis (Anoplura)

The ultrastructure of the Malpighian tubes in human louse Pediculus humanus corporis has been studied. The cells of the Malpighian tubules have the uniform structure: the apical surface is covered with microvilli, the basal plasmatic membrana forms relatively small invaginations. The microvilli are most developed in cells of the proximal department of the Malpighian tubules. Microvilli of the apical surface of the cells do not contain mitochondria which are localized mainly in supranuclear part of the cell. Cells are lined with a homogenous basal membrane.     

An ultrastructural study of Dirofilaria immitis infection in the Malpighian tubules of Anopheles quadrimaculatus

An ultrastructural study was conducted of the Malpighian tubules of Anopheles quadrimaculatus, both uninfected and following infection with Dirofilaria immitis. The Malpighian tubules in Anopheles are composed of primary and stellate cells. The primary cells are the predominant cell type and are characterized by the presence of membrane-bound, intracellular, mineralized concretions and large apical microvilli containing mitochondria. Following the infective blood meal, the microfilariae enter the primary cells of the Malpighian tubules and reside in the cytoplasm in a clear zone without a delimiting membrane. Cells in infected tubules differ from those in uninfected tubules in that the membranes of the vacuoles surrounding the concretions are disrupted in many specimens. The apical and basal cell membranes and the mitochondria associated with these are not disrupted during the first 6-8 days of infection. These observations differ sharply from those previously described in Aedes taeniorhynchus infected with D. immitis. The observations are consistent with the hypothesis that the extended transport capacity observed in previous physiological studies of An. quadrimaculatus infected with D. immitis are dependent on the prolonged normal ultrastructure of the apical microvilli, mitochondria, and basal membranes.     

Fine structure of the Malpighian tubules of chironomus larva in relation to glycogen storage and fate of hemoglobin

The larval Malpighian tubules of Chironomus tentans were studied using light and electron microscopy. The tubules are composed of two cell types: primary and stellate cells. Both cell types lack muscles, tracheoles, and laminate crystals in the cytoplasm and mitochondria in the microvilli. The primary cells exhibit long, wide basal membrane infoldings associated with mitochondria. They have a number of canaliculi and long, closely packed microvilli. The stellate cells possess shorter interconnecting basal infoldings and shorter, well-spaced microvilli. Both cell types are linked by septate and gap junctions. They have cytoplasmic processes and pedicels which enclose narrow slits between them and that are apposed to a basal lamella. In the 'fed' larva, the cells are stuffed with glycogen which is depleted in the 'starved' larva. Both cell types are involved in the vesicular transport of biliverdin. The presence of coated vesicles, tubular elements and various forms of lysosomes in the primary cells suggests they transport and break down functional hemoglobin. Structural modification of basal infoldings, canaliculi and microvilli is strongly correlated with increased secretory activity of the Malpighian tubules in 'fed' versus 'starved' larva.     

Cytoskeletal F-actin patterns in whole-mounted insect Malpighian tubules.

The distribution of actin filaments in Malpighian tubules of the fleshfly Sarcophaga bullata (Parker) was investigated before and after metamorphosis by means of the rhodamine phalloidin staining method. The numerous primary cells show a pattern of thick basal actin bundles resembling stress fibres of cultured cells, while the apical microvillar zone shows a bright and homogeneous labelling. The less abundant stellate cells contain no such basal actin bundles and their apical microvillar zone gets only faintly stained. Late larval stages display fingerlike infoldings and an increased actin filament concentration at the apical membrane of the stellate cells. During metamorphosis the Malpighian tubules dedifferentiate and eventually redifferentiate to give rise to adult tubules resembling larval ones. The different types of actin filament organisation in the primary and stellate cells of the Malpighian tubules are discussed.     

Anatomy and fine structure of the alimentary canal of the spittlebug Lepyronia coleopterata (L.) (Hemiptera: Cercopoidea)

The alimentary canal of the spittlebug Lepyronia coleopterata (L.) differentiates into esophagus, filter chamber, midgut (conical segment, tubular midgut), and hindgut (ileum, rectum). The filter chamber is composed of the anterior extremity of the midgut, posterior extremity of the midgut, proximal Malpighian tubules, and proximal ileum; it is externally enveloped by a thin cellular sheath and thick muscle layers. The sac-like anterior extremity of the midgut is coiled around by the posterior extremity of the midgut and proximal Malpighian tubules. The tubular midgut is subdivided into an anterior tubular midgut, mid-midgut, posterior tubular midgut, and distal tubular midgut. Four Malpighian tubules run alongside the ileum, and each terminates in a rod closely attached to the rectum. Ultrastructurally, the esophagus is lined with a cuticle and enveloped by circular muscles; its cytoplasm contains virus-like fine granules of high electron-density. The anterior extremity of the midgut consists of two cellular types: (1) thin epithelia with well-developed and regularly arranged microvilli, and (2) large cuboidal cells with short and sparse microvilli. Cells of the posterior extremity of the midgut have regularly arranged microvilli and shallow basal infoldings devoid of mitochondria. Cells of the proximal Malpighian tubule possess concentric granules of different electron-density. The internal proximal ileum lined with a cuticle facing the lumen and contains secretory vesicles in its cytoplasm. Dense and long microvilli at the apical border of the conical segment cells are coated with abundant electron-dense fine granules. Cells of the anterior tubular midgut contain spherical secretory granules, oval secretory vesicles of different size, and autophagic vacuoles. Ferritin-like granules exist in the mid-midgut cells. The posterior tubular midgut consists of two cellular types: 1) cells with shallow and bulb-shaped basal infoldings containing numerous mitochondria, homocentric secretory granules, and fine electron-dense granules, and 2) cells with well-developed basal infoldings and regularly-arranged apical microvilli containing vesicles filled with fine granular materials. Cells of the distal tubular midgut are similar to those of the conical segment, but lack electron-dense fine granules coating the microvilli apex. Filamentous materials coat the microvilli of the conical segment, anterior and posterior extremities of the midgut, which are possibly the perimicrovillar membrane closely related to the nutrient absorption. The lumen of the hindgut is lined with a cuticle, beneath which are cells with poorly-developed infoldings possessing numerous mitochondria. Single-membraned or double-membraned microorganisms exist in the anterior and posterior extremities of the midgut, proximal Malpighian tubule and ileum; these are probably symbiotic.     

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。     

Mitochondrial Placement and Function in Insect Ion-transporting Cells

The ion-transporting epithelia of insects possess some unusualmorphological adaptations which promote close juxtapositionof mitochondria and the ion-transporting plasma membranes. Aparticularly striking example of this adaptation is providedby the movement of branches of mitochondria into and out ofthe apical microvilli in the Malpighian tubules. In the hemipteranRhodmus prohxus, the microvilli in the resorptive lower tubuleare small and contain no mitochondria during the non-transportingperiod. When ion transport is stimulated,either in vivo or invitro, there is a concomitant growth in microvillar volume andsurface area. In addition, branches of mitochondria enter thesemicrovilli. It has been shown that these mitochondrial movementsare driven by an actinassociated process involving the microvillarcore microfilaments. The stimulation for this movement in vivois the insect diuretic hormone. In the lepidopteran Calpodesethhus, the rates of fluid transport which the Malpighian tubulescan sustain vary during the insect's life stages. Larvae andadults show rapid transport, while pupal Malpighian tubulesshow none. In the larvae and adults, microvilli in the Malpighiantubules are large and contain mitochondria. In the pupae, reducedtransport is associated with mitochondrial retraction and microvillarshrinkage. These ultrastructural changes appear tobe regulatedby the insect's developmental hormones. In the Malpighian tubulesof adult female mosquitoes of the genus Aedes, intracellularinfection by microfiliarial nematodes has hen shown to causemitochondrial retraction and reduced rates of fluid transport.A model is presented which serves to summarize currently proposedmechanisms of membrane and mitochondrial function in the ion-transportingepithelia of insects.     

The control of Malpighian tubule developmental physiology by 20-hydroxyecdysone and juvenile hormone

The control of developmental changes in Malpighian tubule cell structure and fluid secretion by 20-hydroxyecdysone and juvenile hormone in the skipper butterfly Calpodes ethlius were studied using (1) in vitro tissue culture, (2) in vivo injection and topical application and (3) tubule transplantation experiments. At pupation, 20-hydroxyecdysone initiates cell remodelling and switches off fluid secretion in the Malpighian tubules. Juvenile hormone inhibits these alterations provided that treatment is begun on the first day of the last larval stage. In the pupal stage, 20-hydroxyecdysone triggers the differentiation of adult cell structure which culminates in the renewal of fluid secretion. The results show that 20-hydroxyecdysone and juvenile hormone regulate Malpighian tubule function by altering cell structure and are discussed with respect to the hormonal reprogramming of the Malpighian tubule cells during development.     

Morphology and function of Malpighian tubules and associated structures in the cockroach, Periplaneta americana.

This paper describes the different regions of the Malpighian tubules and the associated structures (ampulla, midgut, ileum) in the cockroach, Periplaneta americana. There are about 150 tubules in each insect. Each tubule consists of at least three parts. The short distal region is thinner than the other parts and is highly contractile. The middle region comprises most of the tubule length and is composed of primary and stellate cells. Primary cells contain numerous refractile mineral concretions, while stellate cells have smaller nuclei, fewer organelles, simpler brush border, and numerous multivesicular bodies. Symbiont protozoa are sometimes present within the lumen of the middle region near where it opens into the proximal region of the tubule. The latter is a short region that drains the tubular fluid into one of the six ampullae. These are contractile diverticula of the intestine located at the midgut-hindgut junction. The ampulla is highly contractile, and consists of a layer of epithelial cells surrounding a cavity that opens into the gut via a narrow slit lined by cells of unusual morphology. The proximal region of the tubule and the ampulla resemble the midgut in that they have similar micromal origin and reabsorptive function for the proximal region of the tubule and for the ampulla. A number of inclusions found within the tubule cells are described, including peroxisomes and modified mitochondria. Current theories of fluid transport are evaluated with regard to physiological and morphological characteristics of Malpighian tubules. The possible role of long narrow channels such as those between microvilli and within basal folds is considered, as is the mechanism by which these structures are formed and maintained. Also discussed is the role of peroxisomes and symbionts in the excretory process.     

Developmental changes in Malpighian tubule fluid transport

Developmental changes in Malpighian tubule fluid transport were studied using in vitro and in situ preparations from timed larval, pupal and adult skipper butterflies (Calpodes ethlius). The ability to transport fluid and the rate of fluid secretion depend on the developmental stage and the physiological state. Larval tubules are permanently switched on, do not require a diuretic hormone and transport fluid at an increasing rate as the larvae feed and grow. Fluid transport continues at larval-larval moults but is switched off 24 hr before pupal ecdysis. No secretory activity occurs during the first half of the pupal stage when the tubules are remodelled for adult function. The resumption of fluid transport midway through the pupal stage is in preparation for a rapid diuresis at adult emergence. High rates of fluid secretion are associated with feeding or drinking in adults.     

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.     

Fine structure of larval malpighian tubules and rectal sac in the tick Ornithodoros (Pavlovskyella) erraticus (Ixodoidea: Argasidae)

The fine structure of the Malpighian tubules (Mts) and rectal sac (rs) is described in the larval tick Ornithodoros (Pavlovskyella) erraticus before and after feeding up to molting. Mts consist of structurally different pyramidal and cuboidal cells along the entire length of the tubule. In unfed ticks, the two types of cell are characterized by apical microvilli and a few basal membrane infoldings. The abundant pyramidal cells contain glycogen particles, lipid droplets, lysosomelike structures, and rickettsialike microorganisms. After feeding but before molting, pyramidal cells loose glycogen particles and become very dense and dramatically reduced in size. These cells are possibly involved in the formation of guanine crystalloids as an excretory product. In contrast, cuboidal cells, filled with glycogen particles, free ribosomes, and mitochondria in unfed larvae, grow steadily after feeding; their cytoplasm becomes rich in lipid droplets in addition to showing an increase in glycogen particles. Lipid and glycogen could be the source of energy required for water and ion reabsorption in which cuboidal cells are probably involved. The paired-lobe rs consists of one type of cuboidal cells with basal membrane infoldings and a brush-border microvilli covered by a fuzzy coat of glycocalyx. These cells grow rapidly after feeding; they have functional features indicating extensive, selective reabsorption of essential components from excretory products.     

Ultrastructure of osmoregulatory organs in larvae of the brackish-water mosquito,Culiseta inornata (Williston)

The ultrastructure of the Malpighian tubules, ileum, rectum, anal canal, and anal papillae of larvae of the mosquito Culiseta inornata was examined. The Malpighian tubules, rectum, and anal papillae have many of the ultrastructural features characteristic of ion transport tissues, i.e., elaboration of the basal and apical membranes and a close association of these membranes with mitochondria. The Malpighian tubules possess two cell types, primary and stellate. The larval rectum of C. inornata is composed of a single segment containing a homogenous population of cells. In this respect, the larval rectum of C. inornata is distinct from that of saline-water species of Aedes. The cells in the larval rectum of C. inornata, however, closely resemble those of one cell type, the anterior rectal cells, of the saline-water mosquito Aedes campestris with regard to cell and nuclear size, the percentage of the cell occupied by apical folds, and mitochondrial density and distribution. No similarities can be found between the rectum of C. inornata and the posterior segment of the saline-water Aedes, which functions as a salt gland. On this basis, we have postulated that the rectum of C. inornata does not function as a site of hyperosmotic fluid secretion. The ultrastructure of the anal papillae of C. inornata is consistent with a role in ion transport. The significance of these findings to comparative aspects of osmoregulatory strategies in mosquito larvae is discussed.     

An investigation into the effects of inhibitors of fluid production by Locusta Malpighian tubule Type I cells on their secretion and elemental composition

The intracellular elemental concentrations of K, Na, Cl, P, Mg and Ca within Type I cells of the Malpighian tubules of Locusta migratoria have been measured using electron probe X-ray microanalysis. The distribution of Na, K and Cl was not homogeneous within the cells and concentration gradients exist from basal to apical surfaces. The rate of secretion and the cationic composition of the secreted tubule fluid have also been determined. Furosemide (1 mM) inhibited fluid secretion by about 60%, raised the [Na(+)] but did not significantly alter the [K(+)] of the secreted tubule fluid. When Rb(+) replaced K(+) in the saline fluid secretion was also inhibited by about 60%, but no additional inhibition occurred by the simultaneous inclusion of furosemide. Thus, Rb(+) and furosemide probably act at the same transport site, and Rb(+) cannot substitute for K(+) at the basal membrane cotransporter. Bafilomycin (1 μM) dramatically inhibited fluid production by 85%, the [K(+)] of the secreted fluid was reduced by about 30% but the [Na(+)] was almost doubled. Furosemide, in common with other inhibitors of fluid secretion acting at the basal surface (ouabain and Rb(+)), caused a fall in intracellular [K] and a rise in [Na]. Bafilomycin, in common with N-ethyl maleimide, which acts at the apical surface, increased the intracellular [K] but did not affect the [Na].