The fine structure of muscle attachments in a spider (Latrodectus mactans, Fabr.)

The fine structure of a spider myo-apodeme junction is described, and discussed in terms of other arthropod muscle attachments. This is contrasted with the situation in the venom gland, equipped with muscle fibers that control expulsion of the secreted material. The latter involves a cell-free collagenous matrix, lying between the muscle cells and the sheath of the gland. As in other arthropods, skeletal fibers are attached to the apodeme cuticle via specialized epidermal cells, containing oriented microtubules. Interdigitations between these cells and muscle, basally, and cuticle, apically, are described. Extracellular tonofibrillae described elsewhere are inconspicuous in the apodeme cuticle.     

The structure of developing muscle insertions in insects

The relationship between muscles and the components of the integument in muscle insertions have been studied with the electron microscope in two insects, Calpodes ethlius (Hesperiidae, Lepidoptera) and Rhodnius prolixus (Reduviidae, Hemiptera). The area of contact between the muscles and the epidermis is increased by interdigitating processes whose membranes are joined by intermediate junctions. The junctions occur at the level of a Z line so that actin filaments attach directly to them. Within the epidermis, microtubules extend from the junctions of the myoepidermal connection to the cuticle, where they attach to hemidesmosomes which line deep indentations of the membrane. The microtubules probably enable the tendinous epidermal cells to withstand the tensions exerted upon them by the muscles. The epidermis is anchored to the cuticle by tonofibrillae, homogeneous rods secreted in the deep indentations of the plasma membrane. Since the tonofibrillae of successive instars are continous, they penetrate and attach to the cuticulin, the outermost layer of the epicuticle.     

The virgin bursa copulatrix of the butterfly, Calpodes

The different parts of the bursa copulatrix of virgin females of Calpodes ethlius have fine structures consistent with their different functions. The thin pitted cuticle of the bursal sac and its underlying epithelium suggest an absorptive function, with their 'pores' and numerous mitochondria respectively. The highly folded junctional region with its robust muscle and epithelium anchored to the thick folded spinous cuticle by sockets and their tonofibrillae suggest a holding structure capable of great muscular activity. The bursal duct with its hardened cuticle might serve to hold the body of the elongated, cone-shaped sperm sac straight and in position so that its contents might be more easily squeezed out into the seminal duct, whose opening is adjacent to the open end of the sperm sac.     

Ultrastructure of the colon of Locusta migratoria

The ultrastructure of the colon of Locusta migratoria is described. The colon is lined by a thick cuticle that, for the most part, adheres to the underlying epithelium. The cuboid epithelial cells are characterized by moderate invaginations of the apical and, to a lesser extent, basal plasma membranes; the lateral plasma membranes are relatively flat. The bulk of the mitochondria are located in the apical region of the cell and are not particularly associated with any of the plasma membranes. The basal region of the cells contains much rough endoplasmic reticulum, glycogenlike granules, and a predominance of spherical, electron-dense bodies of various sizes. Where muscle fibers make contact with the epithelium, the cells are much reduced; the cytoplasm is usually less electron-dense, and, typically, the nucleus has a thick layer of granular material associated with the inner nuclear membrane. The apical and basal plasma membranes of the reduced epithelial cells contain numerous hemidesmosomes. The apical hemidesmosomes occur in pairs around an extracellular space that contains electron-opaque material. The latter forms tonofibrillae that extend into the endocuticle. Bundles of microtubules are associated with the hemidesmosomes. The tubules traverse the cell from the apical to the basal region. The possible significance of these findings is discussed.     

Hormonal control of tanning by the American cockroach: Probable bursicon mediated translocation of protein-bound phenols

The injection of 2-¹⁴C-dopamine into the haemocoel of the American cockroach at ecdysis results in the binding of diphenols to blood proteins. These same proteins are apparently translocated to the cuticle where the label becomes incorporated into the hard insoluble matrix. On the other hand, if labelled dopamine is injected during the pre-ecdysis period, the radioactivity is found bound to the protein but it is not incorporated into the cuticle. Further investigations have revealed that translocation occurs during the immediate postecdysial phase, which suggests that the epidermal permeability may be mediated by bursicon, the tanning hormone.     

The harvestman tarsus and tarsal flexor system with notes on appendicular sensory structures in laniatores

The tarsal flexor system, a novel system of retinacular structures, is described for the first time based on morphological and ultrastructural examinations of several Neotropical harvestmen (Opiliones: Laniatores). The tarsal flexor system is made up of many individual pulleys that function to maintain close apposition between the tendon and internal ventral surface of the cuticle in the tarsus. Pulley cells are specialized tendinous cells that form the semi‐circular, retinacular pulley system in the tarsus; these cells contain parallel arrays of microtubules that attach to cuticular fibers extending from deep within the cuticle (i.e., tonofibrillae). The tarsal flexor system is hypothesized to provide mechanical advantage for tarsal flexion and other movements of the tarsus. This system is discussed with regards to other lineages of Opiliones, especially those that exhibit prehensility of the tarsus (i.e., Eupnoi). Comparing tarsal morphology of laniatorid harvestmen to other well‐studied arachnids, we review some literature that may indicate the presence of similar tarsal structures in several arachnid orders. The general internal organization of the tarsus is described, and ultrastructural data are presented for a number of tarsal structures, including sensilla chaetica and the tarsal perforated organ. Sensilla chaetica possess an internal lumen with dendritic processes in the center and exhibit micropores at the distal tip. With respect to the tarsal perforated organ, we found no ultrastructural evidence for a sensory or secretory function, and we argue that this structure is the result of a large pulley attachment site on the internal surface of the cuticle. A small, previously undocumented muscle located in the basitarsus is also reported. J. Morphol. 274:1216–1229, 2013. © 2013 Wiley Periodicals, Inc.     

Development of the crystalliferous cuticle of Chamaecyparis lawsoniana (A. Murr.) Pari. (Gupressaceae)

A developmental study of the cuticle has shown that it consists of a homogeneous cuticle proper apposed on the wall and a heterogeneous cuticular layer generated by intussusception of cutin into the wall. At an early stage, the adcrusted cuticle proper is underlain by a ruthenium red-positive layer in which the cuticular layer originates. The origin of the anticlinal flange is referable to an electron-dense, ruthenium red-positive ridge which arises above the anticlinal wall and which also becomes cutinized. At leaf maturity, the inner surface of the cuticular layer, including that of the flange, forms interdigitating protuberances with the cell wall.
Development of the cuticle coincides with deposition of crystals of calcium oxalate in the epidermal cell wall. Initiation of large, early-formed crystals is associated with electron-opaque membranous structures formed close and parallel to the plasmalemma in the young cell wall. Crystals undergo periclinal and anticlinal growth and subsequently become engulfed within the cuticle by development of the cuticular layer. Cutin/polysaccharide interaction during development and the significance of crystal deposition are discussed.     

桃树叶片的角质蒸腾实验

折枝处理降低了叶片下表皮蒸腾速率，提高了上表皮角质蒸腾占叶总蒸腾的比值，使角质蒸腾的上下振幅增大。２ ｍ 高度叶片低于１ ｍ 高度叶片的上表皮角质蒸腾。位于黑纸套内及阴处的叶片上表皮角质蒸腾占叶总蒸腾的比值均增大，而全日上表皮角质蒸腾的振幅却减少。     

Embryonic and post-embryonic development of the Early Cambrian cnidarian Olivooides

Phosphatized specimens of Olivooides from the Early Cambrian of Shaanxi, China, represent a number of developmental stages. These include cleavage, gastrulation, organogenesis, cuticularization, pre-hatching, post-hatching and subsequent growth. This allows the reconstruction of a nearly full developmental sequence of this animal. Olivooides had large (600-870 μm in diameter), sphaerical eggs, indicating a high yolk content. Development was direct. Thus adult characters were forming already in the embryo, and there was no free larval stage. The embryonic development took place within a smooth protective membrane. Gastrulation probably was by polar ingression, and the blastopore appears to correspond to the aperture of the later stages. An embryonic cuticle formed which carried star-shaped structures, stellae, over the entire surface except for a radially folded non-stellate portion around the future aperture. At a later stage, the stellate cuticle was thrown into folds concentric with the aperture. This radially folded tissue then became more dominant. After hatching, the body assumed the shape of a strongly annulated cone, with the stellate cuticle forming the apical part and the folded cuticle forming a longitudinally striate cuticle around the aperture. Subsequent growth took place through the addition of striate tissue. A pentaradial symmetry of the body is suggested by lateral folds in the apical part. Olivooides is interpreted as a cnidarian, probably closely related to the scyphozoans. The conical test may have housed a polyp similar to the thecate polyps of modern coronate scyphozoans, but, unlike the latter, Olivooides had no visible attachment structures. There is no evidence for or against a free medusa stage. The prevalence of lecithotrophic direct developers in the Neoproterozoic and Cambrian, unless reflecting a preservational bias, casts some doubts on evolutionary models that assume larval planktotrophy to be primitive among metazoans.     

AN ULTRASTRUCTURAL STUDY OF VEGETATIVE CELL DIVISION IN OEDOGONIUM BORISIANUM12

Vegetative cell division in Oedogonium borisianum is initiated by the formation of a 3-layered ring adjacent to the wall in the upper portion of the cell. This structure enlarges by the coalescence of vesicles. When the ring is fully developed, the parent wall splits adjacent to the ring, and the ring expands into a cylinder, which becomes the cuticle of the upper daughter cell. The lateral wall then forms between this cuticle and the plasmalemma of the cell. Concurrent with ring development and expansion, the nucleus migrates to a position in the center of the cell and karyokinesis occurs. Commencing with late telophase, evidence of transverse wall formation becomes apparent. The zone between the daughter nuclei contains a layer of microtubules in a plane parallel to the plane in which the transverse wall will develop. Subsequently a random coalescence of vesicles occurs along this plane. During the latter stages of this process, the ring expands and the plane of the transverse wall moves upward to the base of the ring cylinder. The completed transverse wall then fuses at is periphery with the newly formed lateral wall.     

Origin and development of stomatal microanatomy in two species ofEucalyptus

Summary Development of the stomata ofEucalyptus orbifolia (in which they are relatively superficial) andE. incrassata (in which they are deeply sunken) is described from light microscopy of thin sections of resin-embedded material. The envelope of the guard mother cell is retained intact while in the daughter cells (guard cells) the inner and outer thickenings are formed. The mother cell envelope may even remain discrete and intact during early stages of formation of the separation spaces, precursors of the future stomatal pore, between the thickenings. Remnants of the guard mother cell wall may be retained as parts of at least the inner stomatal ledges. Likewise, remnants of the wall which divides the mother cell persist on the maturing guard cells.Sudan III-positive materials, probably cutin, are removed from the cuticle over the mother cell soon after it is formed. The cuticle above the guard cell is finally perforated by enzymic attack forming, inE. incrassata, a large cavity outside the developing stoma into which the outer stomatal ledges grow as extensions of the upper guard cell walls.The termostiole is suggested for the aperture in the cuticle. The flanges of cuticle seen in section to bound it are termedostiolar ledges. The ostiolar ledges are to be distinguished from the outer stomatal ledges, which develop from the upper thickenings of the guard cell initials. The distinction is clear inE. incrassata (and other species with deeply sunken stomata) but not in mesophytic plants or species with superficial stomata such asE. orbifolia in which the outer stomatal ledges are fused with the cuticle.Growth of the outer stomatal ledges inE. incrassata involves transport of wall materials through an annular space, the equivalent of an ectocythode.The relevance of the observations to stomatal development in other genera is discussed.     

Differential mechanism of oxidation of N-acetyldopamine and N-acetylnorepinephrine by cuticular phenoloxidase from Sarcophaga bullata

The mechanism of oxidation of two related sclerotizing precursors—N-acetyldopamine and N-acetylnorepinephrine—by the cuticular phenoloxidase from Sarcophaga bullata was studied and compared with mushroom tyrosinase-mediated oxidation. While the fungal enzyme readily generated the quinone products from both of these catecholamine derivatives, sarcophagid enzyme converted N-acetyldopamine to a quinone methide derivative, which was subsequently bound to the cuticle with the regeneration of o-dihydroxy phenolic function as outlined in an earlier publication [Sugumaran: Arch Insect Biochem Physiol, 8, 73 (1988)]. However, it converted N-acetylnorepinephrine to its quinone and not to the quinone methide derivative. Proteolytic digests of N-acetyldopamine-treated cuticle liberated peptides that had covalently bound catechols, while N-acetylnorepinephrine-treated cuticle did not release such peptides. Acid hydrolysis of N-acetyldopamine-treated cuticle, but not N-acetylnorepinephrine-treated cuticle liberated 2-hydroxy-3′,4′-dihydroxyacetophenone and arterenone. These results further confirm the unique conversion of N-acetyldopamine to its corresponding quinone methide derivative and N-acetylnorepinephrine to its quinone derivative by the cuticular phen-oloxidase. Significance of this differential mechanism of oxidation for sclerotization of insect cuticle is discussed.     

An enzyme from locust cuticle involved in the formation of crosslinks from N-acetyldopamine

Locust cuticle is shown to contain an enzyme activating the β-position in the side chain of N-acetyldopamine. When isolated cuticle is incubated with N-acetyldopamine part of the substrate becomes incorporated into the cuticle and part of it forms soluble reaction products, one of which is identified as a dimer of N-acetyldopamine. In the structure suggested for the dimer both phenolic groups of one molecule of N-acetyldopamine are connected to the β-position of another.     

Cuticle proteins of the boll weevil,Anthonomus grandis,abdomen: Structural similarities and glycosylation

Cuticle proteins are thought to be important in defining the structural and functional differences occurring in insect cuticle. In order to explain and better understand the structural similarities among the cuticle proteins of the cotton boll weevil, Anthonomus grandis Boheman, described in a previous study (Stiles and Leopold, 1990, Insect Biochem.20, 113–125) three series of monoclonal antibody producing hybridoma cell lines were produced. Larval, pupal or adult cuticle proteins were used as antigens. While some of the monoclonal antibodies were specific for one or two cuticle proteins from a single developmental stage, the majority showed multiple cuticle protein binding patterns on Western blots. To determine whether this cross-reaction was due to common oligosaccharide chains bound to the proteins, lectins were used to probe Western blots. Many of the cuticle proteins were found to be glycosylated. The majority of the Con A reactive carbohydrate could be removed from the protein by N-glycosidase F digestion (specific for N-asparagine linked carbohydrate). N-glycosidase F digestion did not reduce the multiple cross-reactions of the monoclonal antibodies, nor did periodate oxidation of the CP. The carbohydrate remaining after enzyme digestion is presumably O-linked to serine/threonine.     

Evolution of striated muscle: jellyfish and the origin of triploblasty

The larval and polyp stages of extant Cnidaria are bi-layered with an absence of mesoderm and its differentiation products. This anatomy originally prompted the diploblast classification of the cnidarian phylum. The medusa stage, or jellyfish, however, has a more complex anatomy characterized by a swimming bell with a well-developed striated muscle layer. Based on developmental histology of the hydrozoan medusa this muscle derives from the entocodon, a mesoderm-like third cell layer established at the onset of medusa formation. According to recent molecular studies cnidarian homologs to bilaterian mesoderm and myogenic regulators are expressed in the larval and polyp stages as well as in the entocodon and derived striated muscle. Moreover striated and smooth muscle cells may have evolved directly and independently from non-muscle cells as indicated by phylogenetic analysis of myosin heavy chain genes (MHC class II). To accommodate all evidences we propose that striated muscle-based locomotion coevolved with the nervous and digestive systems in a basic metazoan Bauplan from which the ancestors of the Ctenophora (comb jellyfish), Cnidaria (jellyfish and polyps), as well as the Bilateria are derived. We argue for a motile tri-layered cnidarian ancestor and a monophyletic descent of striated muscle in Cnidaria and Bilateria. As a consequence, diploblasty evolved secondarily in cnidarian larvae and polyps.     

Comparison between the sclerotization of adult and larval cuticle in Schistocerca gregaria

Femur cuticle from fifth instar larvae of the desert locust, Schistocerca gregaria, has been characterized with respect to composition, rate of deposition, and rate of sclerotization. The results are compared with those from adult cuticle of the same species. The protein compositions of the two types of cuticle are very similar, but the rates of deposition of both protein and chitin are different. The main difference is, however, that sclerotization is restricted to the first day after ecdysis in larval cuticle, whereas in adult cuticle sclerotization continues for at least a couple of weeks. The result is that the endocuticle remains untanned in the larvae, whereas in the adults the whole cuticle becomes tanned.     

Stage-specific Differences in Lectin Binding to the Surface of Anguina tritici and Meloidogyne incognita

The occurrence and distribution of several lectin binding sites on the outer surfaces of eggs, preparasitic second-stage juveniles (J2), parasitic second-stage juveniles (PJ2), females, and males of two tylenchid nematodes, Anguina tritici and Meloidogyne incognita race 3, were compared. In both species, a greater variety of lectins bound to the eggs than to other life stages; lectin binding to eggs was also more intense than it was to other life stages. Species-specific differences also occurred. More lectins bound to the amphids or amphidial secretions of M. incognita J2 than to the amphids or amphidial secretions of A. tritici J2. Lectins also bound to the amphids or amphidial secretions of adult male and female A. tritici, but binding to the cuticle occurred only at the head and tail and was not consistent in all specimens. Canavalia ensiformis and Ulex europaeus lectins bound specifically to the outer cuticle of M. incognita. Several other lectins bound nonspecifically. Oxidation of the cuticle with periodate under mild conditions, as well as pretreatment of the nematodes with lipase, markedly increased the binding of lectins to the cuticle of A. tritici J2 but not, in most cases, to M. incognita J2 or eggs of either species.     

The expansion of Schistocerca gregaria at the imaginal ecdysis: The mechanical properties of the cuticle and the internal pressure

(1) At the imaginal ecdysis of Schistocerca, the cuticle is viscoelastic rather than elastic. (2) The cuticle becomes softer just before emergence. It is suggested that this is due to an ‘eclosion hormone’. The softening permits the extrication of the appendages and an increase in the size of the locust. (3) The stiffness of the cuticle increases transiently at the end of emergence. (4) In the later part of wing expansion, the cuticle becomes elastic and its stiffness again increases. (5) Maximum pressures are recorded about 10 min into emergence, and pressures in the gut are greater than those in the tracheal system. (6) From these results it is concluded that the expansion of the wings initially depends on the high haemolymph pressure and low stiffness. Only in the last 15 min of wing expansion do the processes involved in autonomous expansion become important.     

Ultrastructural changes in intersegmental cuticle during rotation of the terminal abdominal segments in a mosquito

The terminal abdominal segments of male Aedes aegypti rotate 180° within 24 hr after adult emergence, rotation occurring in the intersegmental membrane between abdominal segments VII and VIII. The ultrastructure of this rotating membrane is compared with non-rotating intersegmental membranes at different developmental stages. The deposition of cuticle in both the rotating and non-rotating intersegments appears ultrastructurally similar, and follows the sequential pattern described for other insects. Shortly after adult emergence, however, disruptive changes occur in the membrane cuticle that are more pronounced in non-rotating intersegments. This disruption occurs initially 1 hr after adult emergence and becomes maximal within 3 hr. Disruption appears to occur by the addition of fluid to the cuticle and results in a ten-fold increase in cuticle thickness in non-rotating intersegments but only a two-fold increase in thickness in the rotating intersegment. While in the disrupted condition, the non-rotating intersegmental membranes become extensively folded whereas the cuticle in the rotating intersegment becomes stretched. During rotation, strain forces in the rotating intersegment result in a reorientation of microfibers in the cuticle from parabolic to parallel. This reorientation is presumably brought about by plastic flow.