The anatomy and ultrastructure of the suctorial organ of Solifugae (Arachnida)

Solifugae possess an evertable, adhesive pedipalpal organ (suctorial organ) at the tip of the distal tarsus of each pedipalp that is unique among arachnids. When inverted inside the pedipalp, the suctorial organ is covered with two cuticular lips, a dorsal upper lip and a ventral lower lip, but it can be protruded rapidly in order to facilitate grasping prey or climbing on bushes or even climbing on smooth surfaces due to its remarkable adhesive properties. In this study, the suctorial organs of different species from old world families Galeodidae and Karschiidae and new world families Ammotrechidae and Eremobatidae were investigated by means of light microscopy, scanning and transmission electron microscopy. In all representatives, the suctorial organ is formed by an evertable, cuticular pad with a complex internal stabilizing structure. The procuticle of this pad consists of a lattice-like basal plate and numerous stalked structures connected to this basal plate. The shafts of the stalked structures are regularly organized and ramify apically. The surface of the suctorial organ is constituted of a very thin epicuticle overlaying the ramifying apices forming ridges and furrows on the ventral side of the suctorial organ.     

A sticky situation: solifugids (Arachnida, Solifugae) use adhesive organs on their pedipalps for prey capture

Solifugids (Arachnida, Solifugae) have unique evertable adhesive organs on the tips of their pedipalps, named ‘suctorial’ or ‘palpal’ organs. Previous studies have shown that these organs enable solifugids to climb smooth glass-like surfaces and have hypothesized that these structures facilitate prey capture. Here, we use high-speed videography to demonstrate that the suctorial organs of Eremochelis bilobatus are its primary means of capturing insect prey. We also present calculations of the adhesive pressure exerted by these suctorial organs during real prey capture events.     

Ultrastructure of spermatozoa of Solifuges (Arachnida, Solifugae): possible characters for their phylogeny?

The ultrastructure of spermatozoa is a widely accepted source of characters for phylogenetic studies. In this study the fine structure of sperm cells of representatives of six different New and Old World families (Ammotrechidae, Daesiidae, Eremobatidae, Galeodidae, Karschiidae, Solpugidae) of solifuges (Arachnida, Solifugae) were investigated in order to reveal putative characters suitable for subsequent systematic and phylogenetic analyses. The spermatozoa of solifuges represent a relatively simple type of sperm cells. In general, their spermatozoa are roundish, oval shaped (Ammotrechidae, Daesiidae, Eremobatidae, Solpugidae) or plate-shaped (Karschiidae) with or without membrane protuberances and devoid of a flagellum. Only in Galeodidae, very conspicuous thin and elongated sperm cells occur. The spermatozoa either occur as single cells (Eremobatidae, Solpugidae) or in groups of loose knit cells (Ammotrechidae) or in highly ordered groups (Karschiidae). In contrast to the other families studied here, within the Galeodidae and in the genus Blossia (Daesiidae) sperm cells surrounded by a secretion sheath, clearly representing coenospermia, could be observed.     

Position of Pharyngeal Gland Outlets in Monhysteridae (Nemata)

In this study an attempt was made to determine the position of the outlets and nuclei of the pharyngeal glands in four monhysterid genera. Five Eumonhystera spp., seven Monhystera spp., and eight Monhystrella spp. were studied under the light microscope. Longitudinal sections of an undescribed Monhystera sp. and cross sections of Geomonhystera disjuncta were also studied under the scanning and transmission electron microscope, respectively. The results of the light microscopic studies were inconclusive about the position of the outlets but showed a number of nuclei in the basal part of the pharynx. The scanning and transmission electron microscopic studies revealed five pharyngeal glands and their outlets; their position was as follows: dorsal gland outlet at the base of buccal tooth, first pair of ventrosublateral gland outlets halfway along the pharynx, and second pair of ventrosublateral gland outlets close to the base of the pharynx. It is concluded that at least three, and possibly five, nuclei are in the basal part of the pharynx. This pattern, in the position of the outlets and nuclei, is similar to that in Caenorhabditis elegans (Maupas, 1900) Dougherty, 1953 and may well be the basic plan in the Class Chromadorea (including Secernentia as a subclass).     

Sense organs on palps and fore tarsi of gamasid mites (Parasitiformes, Rhinonyssidae), parasites of the nasal cavity of the Great Tit, the Rock Dove, and the Eurasian Coot

Palpal and tarsal sensilla of mites parasitizing in the nasal cavity of the Great Tit Parus major (Ptilonyssus sairae, Ptilonyssus pari), the Rock Dove Columba livia (Mesonyssus melloi), and the Eurasian Coot Fulica atra (Rallinyssus caudistigmus), were examined under a scanning electron microscope. Differences in the topography of the tarsal organ reflect phylogenetic relations of species and genera, whereas differences in the structure of palpal receptors reflect the ecological peculiarities of parasitism.     

Fine structure of diffused pseudobranchial neurosecretory cells associated with carotid labyrinth in an air‐breathing catfish Clarias batrachus

Fish are known to have branchial chemoreceptors and even extrabranchial chemoreceptors to meet the challenges of aquatic environment. The pseudobranchial neurosecretory system associated with carotid labyrinth (CL) is one such example. CL – a chemosensory organ is well known in amphibians. The homologous structure also exists in fish. Clusters of neurosecretory cells, close to the CL and the first two efferent branchial arteries occur in catfish and a few other groups of teleosts. These cells belong to the pseudobranchial neurosecretory system (PNS). To reveal the ultrastructure of CL and the pseudobranchial neurosecretory cells (PNSCs), environmental scanning electron microscope (ESEM) and transmission electron microscope (TEM) investigations were made in an Asian air‐breathing catfish Clarias batrachus. Under ESEM, the PNS appeared as a mass of cells innervated by nerves and supplied by blood capillaries. The CL appeared to have a network of blood capillaries. The transmission electron microscopic investigations showed pear shaped PNSCs having different sizes of dense cored vesicles (DCVs), numerous mitochondria, nerve varicosities, indicating a secretory function of the cells. The CL shows a close association with PNSCs and smooth muscles. Although the exact function of the CL and associated PNSCs in the biology of fish is far from clear, their morphology suggests they are involved in a stress response such as to hypoxia.     

Analysis of the Anointing and Grooming Behavior of Several Adult Insects in Typhlocybinae (Hemiptera: Cicadellidae)

Anointing and grooming behaviors of the adult of Singapora shinshana (Matsumura) (Erythroneurini), Empoascanara (Empoascanara) mai Dworakowska (Erythroneurini), Austroasca vittata (Lethierry) (Empoascini), and Limassolla diospyri Chou et Ma (Zyginellini) were recorded and analysed. Through observation and analysis of their anointing and grooming, we found that there is a preparation stage before anointing behavior in typhlocybines. Anointing and grooming are repeated several times and the numbers and interval times differ from species to species among the Typhlocybinae. Ultramorphological examination of brochosomes and the morphology of the brochosome field, metathoracic tibiae and the structures on the head, face, mesothorax and pygofer of E. mai were done using a scanning electron microscope. Brochosomes (IBS) on the cuticle of typhlocybine adults were spherical visible above these integument. Characteristics of anointing and grooming behavior, the provide evidence that the brochosome field may function as a storage site in preparation for the next grooming session.     

Developmental characteristics of floral organs and pollen of Chinese cabbage (Brassica campestris L. ssp. chinensis)

Morphological and cytological studies are complementary approaches to understand the molecular mechanisms that regulate floral developmental pathways. To better understand abnormal mutant phenotypes in floral development, we conducted detailed observations and investigations of the morphology, cytology, and cell ultrastructure of wild-type Chinese cabbage (Brassica campestris L. ssp. chinensis Makino and syn. B. rapa ssp. chinensis) flowers when they developed from primordia to anthesis. First, we measured bud and organ length with a stereo microscope and observed the developmental status and characteristics of the floral organs using a scanning electron microscope; then we made thin slices of anthers to observe the developmental stage and characteristics of pollen using an optical microscope; and finally, we made super-thin slices of anthers to observe the ultrastructure of pollen during its development with the aid of a transmission electron microscope. In this study, the floral developmental continuum was divided into 17 stages based on significant changes in the shape of floral primordia, and the pollen developmental continuum was divided into 14 stages based on the developmental characteristics. The results could provide the morphological basis for further research on the molecular mechanisms that regulate development of the floral organs and/or pollen of Chinese cabbage and their allied species.     

Combining confocal laser scanning and transmission electron microscopy for revealing the mastax musculature in Bryceella stylata (Milne, 1886) (Rotifera: Monogononta)

The rotiferan jaw apparatus (mastax) is characterized by enormous plasticity and according to morphology and feeding strategy, different mastax types can be distinguished. The cuticular hard parts (trophi) of the mastax are often highly specialized and have both a major taxonomic and phylogenetic relevance. Owing to numerous light and scanning electron microscopic studies, the morphology of the trophi is well known but only few attempts have been made to analyze the morphology and functionality of the mastax as a whole. Particularly, the complex muscular system connecting the individual trophi elements and moving them against each other was disregarded in the past. Therefore, the subject of the present study is a detailed analysis of the mastax musculature of the proalid rotifer Bryceella stylata using a combination of transmission electron and confocal laser scanning microscopic techniques, previously applied for revealing the somatic musculature in rotifers exclusively. Based on ultrathin serial sections and phalloidin-dyed specimens, a total number of six paired and two unpaired individual mastax muscles have been identified for the modified malleate trophi system of B. stylata. Possibly homologous muscles in other, so far investigated rotifer species are discussed as well as functional considerations of the individual mastax muscles and their interaction when moving the trophi elements are suggested.     

Combining confocal laser scanning and transmission electron microscopy for revealing the mastax musculature in Bryceella stylata (Milne, 1886) (Rotifera: Monogononta)

The rotiferan jaw apparatus (mastax) is characterized by enormous plasticity and according to morphology and feeding strategy, different mastax types can be distinguished. The cuticular hard parts (trophi) of the mastax are often highly specialized and have both a major taxonomic and phylogenetic relevance. Owing to numerous light and scanning electron microscopic studies, the morphology of the trophi is well known but only few attempts have been made to analyze the morphology and functionality of the mastax as a whole. Particularly, the complex muscular system connecting the individual trophi elements and moving them against each other was disregarded in the past. Therefore, the subject of the present study is a detailed analysis of the mastax musculature of the proalid rotifer Bryceella stylata using a combination of transmission electron and confocal laser scanning microscopic techniques, previously applied for revealing the somatic musculature in rotifers exclusively. Based on ultrathin serial sections and phalloidin-dyed specimens, a total number of six paired and two unpaired individual mastax muscles have been identified for the modified malleate trophi system of B. stylata. Possibly homologous muscles in other, so far investigated rotifer species are discussed as well as functional considerations of the individual mastax muscles and their interaction when moving the trophi elements are suggested.     

Scanning and transmission electron microscopy of the subfornical organ of the grass frog (Rana pipiens)

Summary The ventricular surface of the subfornical organ of the frog is made up of ependymal cells with numerous apical microvilli, occasional cytoplasmic protrusions and many vacuoles projecting into the lumen of the third ventricle. Between these cells dendrites of cerebrospinal fluid-contacting neurons reach the ventricle to terminate in bulbous enlargements. In addition, flask-shaped encephalo-chromaffin cells, containing granulated vesicles and aggregates of filaments in their cytoplasm, project into the cerebrospinal fluid. Surrounding the centrally located capillaries are enlarged dendrites and axons of heterogeneous morphology, some of which appear to originate within the subfornical organ, intermingled with dendrites and axons of normal structure. The glial cells in this region, especially the microglial cells, often contain large lipofuscin inclusions, suggestive of degeneration and subsequent phagocytosis of some of the enlarged dendrites and axons. The normally scarce neurosecretory peptidergic axons become more evident and form typical Herring bodies in stalk-transected animals. Neuronal perikarya of varying morphology are predominantly located peripheral to the region of enlarged dendrites and axons. Supraependymal macrophages are particularly numerous on the subfornical organ.Abbreviations used CSF cerebrospinal fluid - SEM scanning electron microscope, scanning electron microscopy - SFO subfornical organ - TEM transmission electron microscope, transmission electron microscopy Supported, in part, by NIH grant NB 07492The skillful technical assistance of J.G. Linner and the secretarial assistance of Ann Gerdom are gratefully acknowledged. The SEM studies were made possible through a grant from the Graduate College of Iowa State University and the use of the SEM facility in the Department of Botany     

Fine structure and primary sensory projections of sensilla located in the labial-palp pit organ of Helicoverpa armigera (Insecta)

The fine structure and primary sensory projections of sensilla located in the labial-palp pit organ of the cotton bollworm Helicoverpa armigera (Insecta, Lepidoptera) are investigated by scanning electron and transmission electron microscopy combined with confocal laser scanning microscopy. The pit organ located on the third segment of the labial palp is about 300 μm deep with a 60-μm-wide opening, each structure containing about 1200 sensilla. Two sensillum types have been found, namely hair-shaped and club-shaped sensilla, located on the upper and lower half of the pit, respectively. Most sensilla possess a single dendrite. The dendrite housed by the club-shaped sensilla is often split into several branches or becomes lamellated in the outer segment. As reported previously, the sensory axons of the sensilla in the labial pit organ form a bundle entering the ipsilateral side of the subesophageal ganglion via the labial palp nerve and project to three distinct areas: the labial pit organ glomerulus in each antennal lobe, the subesophageal ganglion and the ventral nerve cord. In the antennal lobe, the labial pit organ glomerulus is innervated by sensory axons from the labial pit organ only; no antennal afferents target this unit. One neuron has been found extending fine processes into the subesophageal ganglion and innervating the labial palp via one branch passing at the base of the labial palp nerve. The soma of this assumed motor neuron is located in the ipsilateral cell body layer of the subesophageal ganglion. Our results provide valuable knowledge concerning the neural circuit encoding information about carbon dioxide and should stimulate further investigations directed at controlling pest species such as H. armigera.     

Identification and mass analysis of human fibrinogen molecules and their domains by scanning transmission electron microscopy

The domainal substructure and molecular conformation of human fibrinogen have been investigated by evaluating scanning transmission electron microscopic images of freeze-dried or negatively contrasted native fibrinogen (fractions I-4 and I-9), glutaraldehyde-treated fibrinogen, or plasmic core fragments D₁ and E₂. Although some unstained freeze-dried native or glutaraldehyde-treated fibrinogen molecules were relatively compact and even occasionally spheroidal, typical images were elongated symmetrical tridomainal structures 460 Å ± 20 Å in length; frequently they were bent into a variety of elongated though non-linear arrangements. Their identification as monomolecular forms of fibrinogen by scanning transmission electron microscopic mass measurements resolved uncertainties relating to the identity of such objects as single molecules. The central domains of fraction I-4 molecules had a greater mass than those of fraction I-9 (1.01 × 10⁵M_rversus 7.5 × 10 M_r, respectively). This difference accounted for the observed mass difference between fraction I-4 and fraction I-9 molecules (i.e. 3.27 × 10⁵M_rversus 2.97 × 10⁵M_r, respectively) and suggested that the COOH-terminal region of the Aα chain (major portions of which are always absent from fraction I-9 molecules) is situated within the mass integration radius for the central domain. When the COOH-terminal region of the Aα chain was present it appeared in negative stain as a thread-like structure originating between the middle and outer domains and extending toward the central domain, sometimes appearing to wind around the long axis.The outer domains of negatively stained molecules resembled negatively stained images of fragment D₁ and could frequently be resolved into at least two discrete subdomains, forming an oblong structure usually canted at an angle of ～120 ° to 150 ° relative to the long axis. Our findings are consistent with prevailing tridomainal structural models of fibrinogen and suggest that these molecules are flexible and may exist in unfolded configurations, or as relatively compact, partially or completely folded forms.     

Cytochemical characterization of basement membranes in the enamel organ of the rat incisor

Ameloblasts are unique epithelial cells, in that once they have deposited the entire thickness of enamel and the process of maturation begins, they reform a basal lamina-like structure at their apical surface. In order to characterize further this basal lamina, its composition was analysed using (1) lectin-gold cytochemistry for glycoconjugates, (2) high-iron diamine (HID) staining for sulfated glycoconjugates and (3) immunogold labeling for collagen type IV and laminin. The labeling patterns were compared to that of other more typical basement membranes found in the enamel organ. Sections of rat incisor enamel organs embedded in Lowicryl K4M were stained with Helix pomatia agglutinin (HPA), Ricinus communis I agglutinin (RCA), wheat germ agglutinin (WGA) and Ulex europaeus I agglutinin (UEA). Samples from the late maturation stage were also reacted en bloc with lectins and embedded in Epon for transmission electron microscopic examination or prepared for scanning electron microscopy. Such samples were also stained with HID and conventionally processed for Epon embedding. Tissue sections were then reacted with thiocarbohydrazide-silver proteinate (TCH-SP). Analysis of the lectin labeling suggested that the region of extracellular matrix immediately adjacent to ameloblasts, where the basal lamina is situated, was intensely reactive with HPA and RCA, moderately reactive with WGA, and weakly reactive with UEA. In general, other basement membranes were mildly reactive with all lectins used. No HID-TCH-SP staining was observed directly over the basal lamina while numerous stain deposits were present over other basement membranes of the enamel organ. Immunolocalization of collagen type IV and laminin yielded a weak and variable labeling over the basal lamina. These results are consistent with the concept of basement membrane heterogeneity and, although the precise nature and composition of the basal lamina associated with maturation stage ameloblasts remain to be determined, they suggest that it may possibly function as a specialized basement membrane with particular compositional characteristics.     

Electron microscopy of the surfaces of bacillus spores

The surface structures of the spores of Bacillus cereus, Bacillus thuringiensis, and Brevibacillus laterosporus were studied by transmission and scanning electron microscopy. Platinum deposition and negative staining with uranyl acetate revealed appendages and exosporium in B. thuringiensis and B. cereus. The exosporium structure was visualized by negative staining and ultrathin sectioning. For staining the exosporium polysaccharide, Alcian blue was used during fixation. The results obtained show the differences in structural organization of appendages and exosporium in different strains. Canoe-shaped inclusions were revealed in all Br. laterosporus strains, while strain IGM16-92 had a fibrillar capsule as well. Electron microscopy using a dual beam scanning electron microscope Quanta 200 3D provided the information of the spore surface relief without sample treatment (fixation and dehydration). The spores of Br. laterosporus strains had folded surface, unlike the smooth surface of B. cereus and B. thuringiensis spores. The diversity of external spore structures was shown within a species, which may be used for detection of bacteria at the strain level. Optimized procedures for visualization of spore surface by different electron microscopic techniques were discussed.     

THE ULTRASTRUCTURE OF PHALACROCLEPTES VERRUCIFORMIS, AN UNCILIATED CILIATE PARASITIZING THE POLYCHAETE SCHIZOBRANCHIA INSIGNIS

The organization of Phalacrocleptes verruciformis is, in general, less complex than that of other ciliates, and no kinetosomes have been observed. However, there are numerous suctorial tentacles at the surface of the body, and the pellicle is characterized by close-set villus-like projections. The tentacles are very small (about 430 mµ in length, and about the same in diameter), but show the essential features of tentacles of suctorians such as Tokophrya, Podophrya, and Ephelota. Each tentacle is reinforced by eight pairs of fibrils arranged concentrically just within its wall, and contains a single missile-like body (MLB). The tentacles become attached to the cilia of the host, and serve for feeding upon the plasmatic contents of the cilia as well as for maintaining contact with the host. The MLB's originate in the endoplasm, and then migrate toward the surface and become incorporated into the tentacles. When feeding is initiated, the membrane covering the outermost nozzle-like portion of the MLB becomes continuous with the membrane of the cilium, and there are other changes in the structure of the MLB which suggest enzymatic activity. Although it appears that Phalacrocleptes is a suctorian, the complete absence of kinetosomes sets this organism apart from other members of the group.     

Method: automatic segmentation of mitochondria utilizing patch classification,contour pair classification,and automatically seeded level sets

Background  

While progress has been made to develop automatic segmentation techniques for mitochondria, there remains a need for more accurate and robust techniques to delineate mitochondria in serial blockface scanning electron microscopic data. Previously developed texture based methods are limited for solving this problem because texture alone is often not sufficient to identify mitochondria. This paper presents a new three-step method, the Cytoseg process, for automated segmentation of mitochondria contained in 3D electron microscopic volumes generated through serial block face scanning electron microscopic imaging. The method consists of three steps. The first is a random forest patch classification step operating directly on 2D image patches. The second step consists of contour-pair classification. At the final step, we introduce a method to automatically seed a level set operation with output from previous steps.     

Plant epicuticular lipids: alteration by herbicidal carbamates

The effect of several carbamates and trichloroacetic acid on the biosynthesis of epicuticular lipids from leaves of pea (Pisum sativum) was tested by chemical and visual methods. The carbamates tested included S-(2,3-dichloroallyl) diisopropylthiocarbamate (diallate), N-(3-chlorophenyl) isopropylcarbamate (chloropropham), S-ethyl dipropylthiocarbamate, and 2-chloroallyl diethyldithiocarbamate. Diallate reduced epicuticular lipids by 50% when the plants were root-treated and by 80% when vapor-treated. These results were supported by scanning electron microscopy and carbon replica techniques with transmission electron microscopy. The ratio of wax lipid components in the diallate-treated plants remained unchanged, with the exception of the primary alcohols, which were reduced. Diallate appears to interfere with the biosynthesis of a precursor to the elongation-decarboxylation pathway of lipid synthesis. N-(3-Chlorophenyl)isopropylcarbamate had no significant effect on total amounts of extractable epicuticular lipids, nor did it alter the structure of the wax formation on the leaves. The scanning electron microscopy micrographs indicated that S-ethyl dipropylthiocarbamate significantly reduced wax formation on pea leaves. 2-Chloroallyl diethyldithiocarbamate altered the structure of the wax formations, but not the total amount of wax (scanning electron microscopy). Trichloroacetic acid had little effect on wax deposition compared to diallate or S-ethyl dipropylthiocarbamate (scanning electron microscopy). The implication of the effect of the carbamates on epicuticular lipids and penetration of subsequent topically applied chemicals is discussed.