Morphological,ultrastructural, histochemical and electrophoretic studies on the venom system of Nasonia vitripennis walker (hymenoptera: Pteromalidae)

The venom system of Nasonia vitripennis is well-developed and composed of an unbranched acid gland and associated reservoir. Fine-structural, histochemical and electrophoretic studies indicate that the venom is produced by two protein-secreting epithelia. The bulk of the venom is synthesised in the columnar cells of the acid gland and discharged via “vesicular organelles” and the efferent ductular system into the lumen of the reservoir. The acid gland also contains squamous chitogenous cells, situated either around the central lumen or interposed between the bases of the columnar cells. Once within the reservoir, the venom is probably activated by enzymatic secretions from the reservoir secretory cells. Each of these cells has a “vesicular organelle” but, in contrast to the columnar cells of the acid gland, the cytoplasm contains a preponderance of free ribosomes, and protein segregation apparently occurs outside the Golgi complexes. The venom is expelled through the efferent discharge duct by muscular contractions, which open the duct lumen and bring it into contact with the funnel of the ovipositor. Excessive distortion of the duct is prevented by a massive ventral ligament.     

A fine structure study of venom gland cells in globiferous pedicellariae from Stronglocentrotus purpuratus (Stimpson)

Cells in secretory glands of globiferous pedicellariae from Strongylocentrotus purpuratus (Stimpson) were studied with the electron microscope and subjected to preliminary light microscopic, histochemical analysis. Specimens for electron microscopic observation were fixed with chilled 2% glutaraldehyde in sea water postfixed in cold 1.33% osmic acid, and embedded in Araldite 502 epoxy resin Samples for histochemical analysis were fixed in the same manner, and then embedded in n-butylmethacrylate. Secretory cells line and fill partially bifurcated, muscular gland sacs located peripherally on each of three jaw elements comprising the pedicellarial head. Cells from venom glands are typically mucoid in appearance, possessing small volumes of basally displaced, vesiculated cytoplasm and an extensive system of vacuoles dominating the apical nine-tenths of each cell. These vacuoles enclose ground substances of various densities and staining affinities. Despite their extensive vacuolation, gland cells contain numerous cytomembrane complexes indicating metabolic activity just prior to fixation. Deciduous endoplasmic reticulum, Golgi complexes, large vacuoles, and various species of vesicles associated with these membrane systems are found in spatial proximity which indicates an apparent biosynthetic association. Preliminary histochemical tests on sections embedded in acrylic plastic indicate vacuolar products may consist of protein and nonsulfated acid mucosubstances. Gland cells are probably holocrine in function, releasing their vacuolar complement upon constriction of the muscular gland sac. There is no evidence indicating delivery of non-membrane bounded, granular secretion to an acellular lumen within the gland sac.     

腰带长体茧蜂毒液器官和卵巢的形态学及其超微结构

应用超薄切片和电镜技术,观察内寄生蜂腰带长体茧蜂Macrocentrus cingulum Brischke毒液器官和卵巢的形态结构。腰带长体茧蜂毒液器官由1个毒囊和2条毒腺组成,毒腺接于毒囊的顶端。毒腺由单层分泌细胞、退化的外胚层细胞和环腔的内膜构成,分泌细胞主要由1个明显的细胞核和1个较大囊状细胞器构成,囊状细胞器的功能是分泌毒液。毒囊由肌肉鞘和扁平细胞层构成,但没有分泌细胞。腰带长体茧蜂卵巢1对,每个卵巢由10条左右卵巢小管组成,与侧输卵管相接处略微膨大形成卵巢萼区。2条侧输卵管在产卵管基部会合形成1条总输卵管与产卵管相接。毒液器官通过毒囊的毒液导管附着在总输卵管上。对寄生蜂毒液器官的生物学、细胞学及在分类进化上的意义进行研究。     

Morphology and ultrastructure of the venom glands of the northern pacific rattlesnake Crotalus viridis oreganus

The venom gland of Crotalus viridis oreganus is composed of two discrete secretory regions: a small anterior portion, the accessory gland, and a much larger main gland. These two glands are joined by a short primary duct consisting of simple columnar secretory cells and basal horizontal cells. The main gland has at least four morphologically distinct cell types: secretory cells, the dominant cell of the gland, mitochondria-rich cells, horizontal cells, and “dark” cells. Scanning electron microscopy shows that the mitochondria-rich cells are recessed into pits of varying depth; these cells do not secrete. Horizontal cells may serve as secretory stem cells, and “dark” cells may be myoepithelial cells. The accessory gland contains at least six distinct cell types: mucosecretory cells with large mucous granules, mitochondria-rich cells with apical vesicles, mitochondria-rich cells with electron-dense secretory granules, mitochondria-rich cells with numerous cilia, horizontal cells, and “dark” cells. Mitochondria-rich cells with apical vesicles or cilia cover much of the apical surface of mucosecretory cells and these three cell types are found in the anterior distal tubules of the accessory gland. The posterior regions of the accessory gland lack mucosecretory cells and do not appear to secrete. Ciliated cells have not been noted previously in snake venom glands. Release of secretory products (venom) into the lumen of the main gland is by exocytosis of granules and by release of intact membrane-bound vesicles. Following venom extraction, main gland secretory and mitochondria-rich cells increase in height, and protein synthesis (as suggested by rough endoplasmic reticulum proliferation) increases dramatically. No new cell types or alterations in morphology were noted among glands taken from either adult or juvenile snakes, even though the venom of each is quite distinct. In general, the glands of C. v. oreganus share structural similarities with those of crotalids and viperids previously described.     

Histology,histochemistry, and emptying mechanism of the venom glands of some elapid snakes

The venom glands of several species of elapid snakes are described. The main venom gland consists of many tubules which usually contain large amounts of secretion product. The accessory gland surrounds the entire venom duct and is usually composed of uniform mucous epithelium. The epithelium lining the tubules of the accessory gland of Naja naja is composed of two distinct types of cells. Histochemical tests indicate that the main venom gland reacts with mercury bromphenol blue and PAS but not with alcian blue. The accessory gland reacts with PAS and alcian blue, and not with mercury bromphenol blue. Treatment of sections with sialidase demonstrates the presence of a sialomucin in the accessory gland. Stimulation of the muscles associated with the venom gland offers an indication of the venom expulsion mechanism of Bungarus caeruleus. A comparison of the venom apparatus of elapid and viperid snakes emphasizes marked differences in the internal anatomy of the venom glands, muscles associated with the gland, and arrangement of glandular components. The morphological differences and dissimilar venom expulsion mechanisms support the recent view of the polyphyletic origin of venomous snakes.     

The form of the globiferous pedicellarial ossicles of the regular echinoid, Psammechinus miliaris Gmelin.

The form of the globiferous pedicellariae from Psammechinus miliaris is described. The valve ossicle resembles that of Parechinus in its triangular valve shape and open blade form, contrasting with Echinus in these features. The present study indicates that the relationship of the valve nerve pathway to the skeletal ossicle is related to the form of the distal blade. It is proposed that subterminal teeth or a tubular blade may be alternative structural devices for strengthening the venom tooth during venom injection.     

Fine structure and presumed functions of the pedicellariae of Echinocardium cordatum (Echinodermata,Echinoida)

Summary Tridactylous, trifoliate, and globiferous pedicellariae occur on the body surface of Echinocardium cordatum. Tridactyles have three forms: the typical, the rostrate, and the large forms. Both typical and rostrate tridactyles and trifoliates occur all around the echinoid body (trifoliates are, however, 4 times more numerous than tridactyles). Large tridactylous and globiferous pedicellariae are restricted to the peribuccal area.As a general rule tridactyles and trifoliates are similar in morphology. The distal part of the valves forms an open blade and bears lateral teeth and/or denticles (single or in combs). The stalk consists of a rigid proximal part supported by an axial rod and a flexible distal part which includes an axial fluid-filled cavity. The cavity is surrounded by muscle fibers and acts as an hydroskeleton, allowing the undulating-coiling movements of the flexible part of the stalk. Trifoliates are always active while tridactyles react only to direct or indirect mechanical stimulation.The valves of the globiferous pedicellariae have a tubular distal part whose upper opening is surrounded by teeth. There is no differentiated venom gland but a cluster of epithelial glandular cells located at the level of the valve upper opening. A small ciliary pad occurs just below the glandular cluster. Globiferous stalks are not flexible, being supported for their full length by an axial rod. Globiferous pedicellariae appear to be sensitive only to chemical stimulation.The presumed functions of E. cordatum pedicellariae are (1) cleaning of the body surface and ciliary structures (trifoliates), (2) protection against sedimenting particles (tridactyles), and (3) defense of the peribuccal area against potential small predators (globiferous pedicellariae).     

Development of the venom ducts in the centipede Scolopendra: an example of recapitulation

In contrast to previous claims that (a) there is a law of recapitulation and, conversely, (b) recapitulation never happens, the evolutionary repatterning of development can take many forms, of which recapitulation is one. Here, we add another example to the list of case studies of recapitulation. This example involves the development of the venom claws (forcipules) in the centipede Scolopendra subspinipes mutilans, and in particular the development of the duct through which venom flows from the gland that produces it (proximal) to the opening called the meatus (distal) through which it is injected into prey. Most of the information we present is from early postembryonic stages—these have been neglected in previous work on centipede development. We show that the venom ducts arise from sutures that are invaginations of the cuticle. In S. s. mutilans, the invagination in each forcipule forms into a tubular structure that detaches itself from the exoskeleton and moves toward the center of the forcipule. This is in contrast to extant Scutigera, and also, probably, Scolopendra's extinct Scutigera‐like ancestors, where the duct remains attached to the cuticle of throughout development. Thus, S. s. mutilans exhibits a recapitulatory repatterning of development.     

The mechanism of venom secretion from Duvernoy's gland of the snake Thamnophis sirtalis

The venom glands of snakes of the families Elapidae and Viperidae are thought to have evolved from Duvernoy's gland of colubrid ancestors. In highly venomous snakes elements of the external adductor musculature of the jaw insert fibers directly onto the capsule of the venom gland. These muscles, upon contraction, cause release of contents by increasing intraglandular pressure. In Thamnophis sirtalis, a colubrid, there is no direct connection between Duvernoy's gland and the adductor musculature. The anatomical arrangement of the gland, skull, adductor muscles, and the integument is such that contraction of the muscles may facilitate emptying of the gland. This hypothesis was tested by electrical stimulation of the muscles, which resulted in significantly greater release of secretion than elicited by controls. The results suggest a possible early step in the evolution of a more intimate association between venom glands and adductor musculature in highly venomous snakes.     

Ultrastructure of venom glands in the frog (Rana esculenta)

Electron microscopic study of skin venom glands in the frog, Rana esculenta, revealed the syncytial structure of the inner (secretory) wall which presents two distinct zones: a basal (juxtamuscular) one, which contains nuclei and major cytoplasmic organelles, and an apical one where large electron-dense granules form and accumulate. Granules are seen to arise inside clusters of tightly packed smooth endoplasmic reticulum (SER) elements, which suggests that the SER system is mainly involved in synthesis of this material. A high glutaraldehyde concentration (5%) also reveals a poorly defined material filling the intergranular cytoplasm. No apical limits to the syncytium could be traced, which suggests massive holocrine secretion. Nerves insinuate between the muscle cells and occur all along the internal face of the muscular layer, sometimes in close contact with the syncytium. The gland duct, the wall of which consists of epidermal cells, is blocked, in contact with the gland, by an epidermal bud linked externally to the muscle layer surrounding the gland. Thus, only strong muscle tension such as to expel all or part of the epidermal bud can trigger granule release. This phenomenon can be induced by the subcutaneous injection of epinephrine, but the high and distressing dose needed to provoke appreciable changes in venom glands renders unlikely any natural intense venom release triggered by epinephrine in the frog.     

Immunocytochemical localization and secretion process of the toxin CSTX-1 in the venom gland of the wandering spider Cupiennius salei (Araneae: Ctenidae)

Fluorescein and horseradish peroxidase-labeled monoclonal antibodies were used to localize the predominant toxic peptide CSTX-1 in the venom gland of the spider Cupiennius salei. There was no polarity of CSTX-1 expression in repleted glands, whereas the glands of previously milked spiders showed a decreasing immunofluorescent response from the distal to the proximal portion. Detailed investigation revealed a new structure in the venom-secreting epithelium, which is postulated to be an evolutionary adaptation to increasing gland volume. CSTX-1 was found to be synthesized and stored as a fully active toxin within complex units, composed of long interdigitating cells running perpendicular to the muscular sheath and extending into the central lumen of the gland. These venom-producing units were found in all sectors of the gland, including the transitional region between the main gland and the venom duct. The venom is liberated from the venom-producing units into the glandular lumen following the contraction of the surrounding muscle layer. Free nuclei or other cellular fragments, which would have provided evidence for a holocrine secretion process, were not found in the glandular lumen or in the crude venom obtained by electrical stimulation. The fine regulation of the spider's venom injection process is postulated to be the function of the bulbous ampulla, situated in the anterior third of the venom duct.     

Morphology and ultrastructure of a secretory region enclosed by the venom reservoir in social wasps (Insecta,Hymenoptera)

The morphology and ultrastructure of the convoluted gland inside the venom reservoir of four species of social Vespidae are described. The cells of the venom gland (including the convoluted gland) can be divided into six groups: (1) epithelial cells, (2) glandular cells with the end apparatus secreting into the tubule inside the convoluted gland (internal or embedded tubule), (3) a continuous arrangement of glandular cells with the end apparatus secreting directly into the venom reservoir, (4) glandular cells that are loosely dispersed along the tubule lumen between the free tubules and the embedded tubule of the convoluted gland, (5) secretory cells of the free tubules and (6) duct cells. One kind of secretory cell, hitherto unknown and described in this paper (group 3), is characterized by the presence of a well-developed end apparatus, usually with enlarged extracellular spaces, but lacking the normally associated duct cells. The secretory cells contain several stacks of granular endoplasmic reticulum, but these are mainly concentrated in the middle of the cell. The basal half of the cells contains many lipid droplets. Although the function of the convoluted gland is not yet understood, an hypothesis is related to what is known of the function of reservoir secretory cells in solitary wasps. All wasp species studied showed the same organization of the convoluted gland, which clearly distinguishes their venom gland from that of Sphecidae.     

Ultrastructure of maxillary gland of Antrobathynella stammeri (Syncarida,Malacostraca)

The maxillary gland of the highly adapted stygobiont species, Antrobathynella stammeri (Bathynellacea, Syncarida), consists of an end sac, an excretory tubule, and a terminal duct. No valve was found. The excretory tubule forms a loop extending back into the fourth thoracic segment. The end sac is composed of five typical podocytes. Ultrastructurally and functionally, two cell types characterize, respectively, proximal and distal sections of the excretory tubule. Epithelial cells are covered with extremely broad (up to 0.4 μm) microvilli. A basal labyrinth was not seen. Therefore, it is unlikely that the maxillary gland is able to produce a hypoosmotic urine necessary for freshwater animals. Tubule cells can be surrounded by parenchymal cells that produce numerous vesicles, suggesting possible physiological interactions between tubule cells and parenchyma. The ectodermal terminal duct is lined with cuticle and is differentiated into a pulsatile body consisting of two interconnected ampules. The first functions as a bladder. The second ampule, under muscular control, excretes the urine. Pulsatile body, looping tubule, and broad microvilli appear to be distinctive features of the bathynellacid excretory organ. © 1996 Wiley-Liss, Inc.     

后毒牙类毒蛇

长期以来,人们仅把具有沟牙和管牙的蛇视为毒蛇,然而,近年来发现游蛇科中的虎斑颈槽蛇、红脖颈槽蛇、颈棱蛇、赤链蛇等既无管牙,也无沟牙,却频频发生这类蛇咬伤人后引起中毒的事例,甚至出现被咬伤致严重出血休克死亡的事件。经深入研究后发现,这些蛇虽没有沟牙和管牙,但却具有产生毒性分泌物的毒腺—杜氏腺(Duvernoy′s gland)及皮下腺,且不同的毒腺具有不同的毒性作用,可表现为出血不止、溶血、呼吸困难、肾损害等。这类蛇与毒腺的导管有联系的上颌牙明显粗大,上颌牙与上颌骨、横骨连接牢固,毒腺里的毒液可顺着粗大的上颌牙流入伤口,因此,应视为"后毒牙类毒蛇"。     

Anatomical correlates of venom production in Conus californicus

Like all members of the genus, Conus californicus has a specialized venom apparatus, including a modified radular tooth, with which it injects paralyzing venom into its prey. In this paper the venom duct and its connection to the pharynx, along with the radular sac and teeth, were examined using light and transmission electron microscopy. The general anatomy of the venom apparatus resembles that in other members of the genus, but several features are described that have not been previously reported for other species. The proximal (posterior) quarter of the venom duct is composed of a complex epithelium that may be specialized for active transport rather than secretion. The distal portion of the duct is composed of a different type of epithelium, suggestive of holocrine secretion, and the cells display prominent intracellular granules of at least two types. Similar granules fill the lumen of the duct. The passageway between the lumen of the venom duct and pharynx is a flattened branching channel that narrows to a width of 10 micro m and is lined by a unique cell type of unknown function. Granular material similar to that in the venom duct was also found in the lumen of individual teeth within the radular sac. Mass spectrometry (MALDI-TOF) demonstrated the presence of putative peptides in material derived from the tooth lumen, and all of the more prominent species were also evident in the anterior venom duct. Radular teeth thus appear to be loaded with peptide toxins while they are still in the radular sac.     

Comparative morphology of the foot structure of four genera of Loxosomatidae (Entoprocta): Implications for foot functions and taxonomy

Entoprocta is a group of mostly cryptic, benthic invertebrates with a sedentary lifestyle. Here, we investigate the morphology of the entoproct foot, which is an important structure in attachment and locomotion. We describe the foot structure of four solitary entoprocts, Loxosoma monilis, Loxosomella stomatophora, Loxocorone allax, and Loxomitra mizugamaensis, by means of light and transmission electron microscopy. Gland cells containing secretory granules were found in the foot of all the four species. In L. monilis, the gland cells densely paved the underside of the disc‐shaped foot, but no duct or groove was found. In L. stomatophora and L. allax, a foot gland was present at the frontal end of a foot groove. The foot gland was a solid cell mass in the former species but a sac‐like structure in the latter. Two types of groove accessory cells were recognized in both species; groove bulge cells (GBCs) showed large cytoplasmic bulges extending into the groove lumen, while groove microvillus cells have microvillus mats in the lateral wall of the groove. The bulges of GBCs in L. stomatophora are slender and attached to one another with desmosomes, forming appendages that extend down to the substratum, hinting at their contribution to attachment and locomotion. The bulges in L. allax form large swellings that fill the groove lumen and are connected to the surrounding cells with hemidesmosomes. In the liberated buds of L. mizugamaensis, tripartite gland cell masses were found at the basal end of the stalk, but no groove was found. A small invagination, which may be the opening of the gland, was found at the center of the foot tip, where the liberated buds attach themselves to the substratum and then metamorphose into adults. No openings were found at the lateral terminal wings, which support locomotion in Loxomitra species. J. Morphol. 271:1185–1196, 2010. © 2010 Wiley‐Liss, Inc.     

Morphology and ultrastructure of the Dufour gland in workers of social wasps (Hymenoptera, Vespidae)

The Dufour gland in workers of vespine wasps appears as an unpaired tubiform gland that opens in close proximity to the sting base. The epithelial cells that line the central reservoir are characterized by apical microvillus-like projections and deep basal invaginations. Their cytoplasm contains a well-developed Golgi apparatus, numerous mitochondria, as well as strands of smooth endoplasmic reticulum. The Dufour gland duct occurs ventrally to the venom gland duct, and bends downward near the sting base to open in the dorsal vaginal wall. In this region, the duct is dorsoventrally flattened, and shows conspicuous bundles of parallel microtubules in the epithelial cells, that transmit the pulling forces of the myofilaments of the underlying muscular supply to the cuticle. This results in an active opening mechanism regulated by muscular contraction, while passive closure probably results from the return of the cuticular intima to a rest position.     

Morphology,morphometry and ultrastructure of the maxillary gland of the terrestrial isopod Porcellio scaber (Crustacea,Oniscidea)

Summary The ultrastructure of the maxillary gland of the terrestrial isopod Porcellio scaber is described. The gland is composed of an end sac, an excretory duct and a terminal duct which opens by a valve at the base of maxilla 2. An epithelium of podocytes in the end sac enables passive ultrafiltration by haemolymph pressure. The excretory duct shows ultrastructural adaptations to secretion and resorption. SEM micrographs reveal the location and the morphology of the valve at the excretory pore. A model reconstructed from serial sections allowed the calculation of morphometric data of the gland. The ultrafiltration area of the end sac and the area of resorption and secretion of the excretory duct amount to 0.091 and 0.157 cm² per 1 g of fresh body weight, respectively. The total volume of the gland is calculated to be 0.04 mm³ in a specimen of 13.7 mm length. In comparison with the marine species Mesidotea entomon, the relative areas for ultrafiltration and resorption of the gland of P. scaber are more than twice as large as the corresponding areas of the marine species. This relative enlargement of the gland in P. scaber and the form of the valve at the excretory pore are seen as adaptations to terrestrial life.     

Ultrastructure of the venom gland of the scorpion,Centruroides sculpturatus (Ewing)

The venom apparatus of the scorpion, C. sculpturatus (Ewing) was studied with light and electron microscopy. Each of the paired glands is lined by secretory epithelium made up of a single layer of columnar cells. Extensive folding in the epithelial layer creates a primitive acinar gland. The secretory products are either membrane-bound or unbound vesicles with discrete morphologies and are observed in the extruded venom, within the lumen of the gland, and within single secretory cells. The venom apparatus, including connective tissues, nerve cells, and muscle tunic is described and correlations are made with observations in other Athropods.