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.     

Fine structure of the stretch receptor in the bursa copulatrix of the butterfly,Pieris rapae crucivora

Summary A pair of multipolar stretch-receptive neurons were found in the bursa copulatrix of the female cabbage white butterfly, Pieris rapae crucivora. The cell body of each neuron, about 10 m in diameter, lies on the edge of the muscular region in the antero-lateral wall of the corpus bursae. No special accessory structure, such as a receptor muscle, is associated with the neuron. The several dendrites extend radially into the muscle layer. The dendrites are ensheathed except for their terminal tips, and, on their course, they anchor repeatedly on the epithelial cells or the muscle fibers in such a manner that their basement membranes fuse together. While the ensheathed dendrite is usually 0.1–0.2 m in diameter, it often forms 1–2 m varicosities especially at anchor sites, so that it looks like a varicose, or beaded, chain. The varicosities contain a number of mitochondria, but only microtubules are found in the fine interconnecting parts of the dendrite. The naked dendritic tips terminate in the basement membrane of the epithelial cell. The varicosities, as well as naked tips, seem to be important for stimulus transduction in the sensory cell of this type.     

The localization of a peroxidase associated with hard cuticle formation in an insect, Calpodes ethlius stoll, Lepidoptera, Hesperiidae

The distribution of a peroxidase associated with the formation of hard cuticle has been studied in developing larvae of Calpodes ethlius. It occurs in granules in several cell types but is most easily observed in the cells making the proleg spines at the 4th to 5th molt. Light microscopy shows peroxidase in numerous granules about 0.5mu in diameter at the time the cuticle of the spine shaft is being deposited. Electron microscopy shows these granules to be multivesicular bodies with peroxidase in the matrix. Peroxidase is also found in cisternae of the rough ER near Golgi complexes, in vesicles of Golgi complexes and in the secretory vesicles which discharge to make cuticle at the apical surface. The cuticle above the plasma membrane where peroxidase is being deposited reacts with DAB in the absence of hydrogen peroxide. Presumably this cuticle has been 'peroxidized' as a first stage in stabilization by cross-linking. Some of the peroxidase secreted at the apical surface is pinocytosed and transported to the multivesicular bodies, suggesting that there may be a precise control of the cuticular environment through the turnover of its soluble components.     

Studies on the silk glands of Calpodes ethlius stoll, (Lepidoptera,hesperiidae)

Each silk gland of Calpodes ethlius consists of five distinct regions: the duct, the green, anterior, middle and posterior regions. Although the gland increases approximately tenfold in length during the larval life, the number of cells remains constant with a concomitant increase in ploidy which is not constant either throughout larval life or in the different regions of the gland. Histochemistry on the glands of the mid-fifth instar larva shows that progressively more mucosubstances are deposited in the lumen, so that while in the distal regions there is only one weakly acidic deposit, this is increased to three more acidic bands in the proximal regions. These bands can be correlated with materials of different electron density. All five regions have characteristic secretory ultrastructure, with prominent secretory vesicles or granules and microvilli. However, the posterior and middle regions have electron-translucent vesicles and relatively short microvilli, while the other three regions have electron dense granules and a more complex, microvillate apical surface. This complexity is greatest in the duct which suggests that it may function in water reabsorption.     

The structure and function of the bursa copulatrix of the monarch butterfly (Danaus plexippus)

The bursa compulatrix of the Monarch butterfly was investigated utilizing light microscopy, histochemistry, and scanning and transmission electron microscopy in order to relate its morphology to the release of sperm from the spermatophore. The bursa has a row of large chitinous teeth on either side of the organ. The dorsal and ventral surfaces are covered with chitinous plates, the plates having bristles on one side. A single layer of cells lies under both the plates and teeth, one columnar cell under each plate, one cuboidal cell under each tooth. The toothed area has no muscle cells. However, the dorsal and ventral hemispheres of the bursa each have a crescent-shaped packet of muscle fibers that traverse the organ; there are no longitudinal fibers. Spermatophores with thick walls were found in the bursal lumen. Morphological evidence suggests that the presence of the spermatophores is sensed by the bristles and that the packets are opened by contraction of the muscles bringing the large teeth into contact with the spermatophore wall.     

The structure of the haemocytes of Calpodes ethlius (Lepidoptera)

The haemocytes of Calpodes ethlius are described with the light and electron microscopes. Five fine structurally distinct types are distinguishable. However only three of these, the granular haemocytes, sphaerule cells and oenocytoids can be positively identified using a series of histochemical stains on smears and on thick sections of araldite-embedded material. The classification is based entirely on the structural features even though several suggestions concerning their function can be made from their fine structure. Intermediates having features of more than one cell type suggest developmental relationships.     

Structure and formation of the chorion in the butterfly, Calpodes

The hemispherical eggshell of Calpodes consists of a domed dorsal surface of thin inner endochorion and a thick, lamellate, outer exochorion, and a flat bottom of predominantly unlamellated endochorion. The endochorion is traversed by small pores and the exochorion by larger ones, which are formed by the withdrawal of processes from the follicular cells. The presence of a phenoloxidase in the ovariolar ducts, lateral and common oviducts and in ovulated eggs, and the stability of ovulated eggs suggest that stabilization of the eggshell is accomplished through quinone tanning. However, the endochorion, which is soluble in sodium dodecyl sulphate (SDS), is more likely cross-linked by di- and tri-tyrosyls through the action of a peroxidase, present in the cells of the ovariolar duct and in the endochorion.     

The origin,transport and cleavage of the molt-associated cuticular protein CECP22 from Calpodes ethlius (Lepidoptera,Hesperiidae)

CECP22 (Calpodes ethlius Cuticular Protein 22 kDa) is a molt associated protein found in the cuticle of C. ethlius larvae and pupae. The mRNA for the CECP22 cuticular protein is expressed in the epidermis and fat body during the intermolt. The protein itself accumulates in intermolt hemolymph, but at molting, when the cuticle is being digested, it is also found in the cuticle of surface integument, tracheae, foregut and hindgut and in the molting fluid. CECP22 exists in two forms. The large form (19.17 kDa, pI 6.2) becomes smaller (16.1 kDa, pI 7.4) by cleavage at the proteolytic cleavage site (position 170) with amidation of the C-terminal. The small, more basic peptide, appears only at molting, first in the cuticle and then in the molting fluid. It is presumed to be the active form of an amidase involved in the earliest stages of cuticle degradation. The inactive form accumulates in the hemolymph during the long intermolt and probably represents an abundant source of precursor enzyme that can be provided to all cuticle containing organs for a precise initiation of cuticle degradation.     

The structure and formation of the cuticulin layer in the epicuticle of an insect, Calpodes ethlius (Lepidoptera, Hesperiidae)

The cuticulin layer is defined as the dense lamina (120–175 Å thick in Calpodes larvae, depending upon the stage) forming the outer part of the epicuticle in insects. It completely invests an insect except for the gut and the openings of some sense organs. It is the first layer to be secreted during the formation of new cuticle. The formation of the cuticulin membrane may be a useful model for studying the origin of membranes in general. It arises as a triple layer de novo and is not a modified plasma membrane. Growth is by accretion at the edges of patches of cuticulin which increase in area until they cover the new surface. The triple layer (i.e. three dense laminae) may develop striations about 30 Å apart transverse to the membrane, which perhaps form a sieve allowing small molecules to pass while protecting the cell from enzymes in the molting fluid. A similar porous structure persists in the tracheoles. After the resorption of molting fluid the triple layered structure again becomes obvious and the outermost layer separates from the other two to become what may be the surface lipid monolayer. The surface patterns in cuticle of various sorts probably arise by buckling of the cuticulin layer as it increases in surface area.     

Microstructure and diversity of the bursa copulatrix wall in Tortricidae (Lepidoptera)

The inner surface and muscle structure of the bursa copulatrix are examined for the first time with Electron Microscopy in some representatives of the family Tortricidae (Lepidoptera). The internal microprotuberances reveal taxon dependent characters unstudied in the Lepidoptera until now. Acanthae occur in almost all taxa studied, whereas ctenidia and papillae are found only in representatives of Tortricinae and Olethreutinae, respectively. Muscles are radially arranged from the signa. Areas of muscle insertion on the bursal wall are not covered by other muscle fibers. Muscle attachments to evaginated areas, e.g. capitulum and diverticulum, suggest an apodeme role for these structures, correlated with long protruding signa. The potential evolutionary and adaptive significance of these new findings are discussed.     

Purification and characterization of three major hemolymph proteins of an insect,Calpodes ethlius (lepidoptera,hesperiidae)

Like many other Lepidoptera, fifth-stage Calpodes larvae have three major hemolymph proteins. Their molecular weights were estimated by 3-15% nondenaturing polyacrylamide gel electrophoresis (N-PAGE) as 470,000 (arylphorin; Ar), 580,000 (storage protein 2; SP2) and 720,000 (storage protein 1; SP1). Carbohydrate is associated with all three, but only Ar has lipid. The three proteins have been purified by preparative N-PAGE and sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. On 3-15% SDS gels, Ar dissociated into 82,000 M_r subunits, SP2 into 86,000 M_r subunits, and SP1 into both 86,000 and 90,000 M_r subunits. The 470,000 M_r protein is identified as Ar because it is rich in aromatic amino acids. The 580,000 and 720,000 M_r proteins are rich in glycine and are called storage proteins. Electron microscopy of negatively stained preparations shows that each polymer has a different geometrical arrangement of subunits. SP1 is a cube made from eight subunits. SP2 is a hexamer in the form of a pentahedral prism. Ar is probably an octahedron made from six subunits. All three geometrical arrangements could permit the presence of a central carrying space.     

The molt/intermolt cycle in the epidermis and other tissues of an insect Calpodes ethlius(Lepidoptera, Hesperiidae)

Tissue responses to the brain and prothoracic glands divide the 5th larval stage of Calpodes into three phases of development, characterized by growth, larval syntheses and pupal syntheses. Tissue specific patterns in the timing of nuclear events fall into one of two categories. In both, the cell number is determined mainly before the 4th to 5th ecdysis. Epidermal cells divide during the second phase coincidentally with an elevated rate of larval cuticle deposition, but in other tissues nuclear replication occurs without division during the first phase so that the cells are polyploid by the time they begin their massive larval syntheses.     

L-Glutamate retrieved with the moulting fluid is processed by a glutamine synthetase in the pupal midgut of Calpodes ethlius

From apolysis until pupal ecdysis, the pharate pupa of the Brazilian Skipper (Calpodes ethlius) lies wrapped in a prepupal shell composed of the larval cuticle and an ecdysial space (ES) filled with enzyme-rich moulting fluid (MF). In the 4h before ecdysis the pharate pupa drinks the moulting fluid through its mouth and anus, and transfers the cuticular degradation products to its midgut (MG). At the same time, extra fluid passes across the body wall of the pharate pupa and flushes out the ES. The MF is recovered at an overall rate of 70μl/h and reabsorbed across the pharate pupal midgut at about 26μl/h. L-Glutamate was found to be the dominant amino acid in the moulting fluid. Total MF glutamate peaked at 850nmol about 8h before pupal ecdysis (P-8), but by ecdysis it had dropped to nearly zero as the MF became diluted with new fluid and was consumed. The drop in glutamate in the ES coincided with a rise in the glutamine content of the fluid in the midgut lumen. The highest rate of glutamine synthesis occurred in midguts isolated from pharate pupae actively drinking MF (P相似文献   

The synthesis of hemolymph proteins by the larval epidermis of an insect Calpodes ethlius (Lepidoptera: Hesperiidae)

Sheets of the dorsal abdominal integument from fifth instar larvae of Calpodes ethlius (Lepidoptera: Hesperiidae) were incubated in artificial hemolymph in the presence of [³⁵S]methionine to investigate protein synthesis and vectorial secretion. The epidermis synthesizes and secretes at least 13 polypeptides basally and 15 apically. Two dimensional analysis of proteins labeled in vitro and in vivo showed that (a) most of the polypeptides secreted on apical and basal surfaces are different, (b) in vitro apical secretions are the same as in vivo cuticular proteins, (c) at least four of the basal secretions can be demonstrated in hemolymph labeled in vivo.Antibodies made against whole hemolymph recognized five basally secreted polypeptides and one apically secreted polypeptide both on fluorograms of immunoprecipitates and immunoblots. Arylphorin is secreted from both surfaces. Arylphorin synthesized in vitro has been identified through its precipitation by antibodies to hemolymph arylphorin in epidermis, cuticle and medium. We conclude that insect epidermis has bi-directional secretion. Cuticular proteins are carried to the apical face. A different set of proteins are carried basally to the hemolymph.