0000000000042526
AUTHOR
August Dorn
Testis differentiation in the glowworm,Lampyris noctiluca, with special reference to the apical tissue
The gonads of Lampyris noctiluca are sexually undifferentiated during the first larval instars. They consist of many gonadal follicles that include the germ stem cells enclosed by the somatic cells of the follicle wall. Follicle wall cells are more numerous at the follicle apices than at the distal parts, but different cell types cannot be distinguished. In male larvae, the appearance of apical follicle tissue, derived from follicle wall cells, marks the onset of testis differentiation. When maximally expressed, the apical tissue occupies about the upper half of the testis follicles and can be observed in larvae of the fifth and sixth instar. The apical tissue is characterized by its “light…
Mesadenes maturation and its hormonal control in the milkweed bug, Oncopeltus fasciatus
Abstract Adult maturation of the mesadenia was examined by electron microscopy. The glandular epithelium exhibits only one cell type with a polarity along the haemocoel-lumen axis. In newly emerged males, the glands show little signs of secretory activity although the narrow lumen contains some material. Dramatic changes commence at day 2. The rER increases drastically, and at day 3 cisternae of the rER are considerably distended and include a fine granulated product. The lumen of the gland widens greatly and stores material of the same appearance as in the rER inclusions. The mode of the secretory process is not clear since membrane bound vesicles are not formed. SDS-PAGE of the secretion …
Effects of azadirachtin on the regulation of midgut peristalsis by the stomatogastric nervous system inLocusta migratoria
A single injection of azadirachtin into nymphal and adult locusts results in a decrease of food intake but not in reduced food utilization. Feeding deterrency is correlated with a gradual inhibition of midgut peristalsis. Gut movement is under the control of the stomatogastric nervous system, especially the ingluvial ganglion, as demonstrated in anin vitro system. Serotonin is the most potent peristalsis-stimulating drugin vitro. Concurrent with the cessation of midgut peristalsis after azadirachtin injection, a depletion of the serotoninergic cell bodies in the frontal ganglion can be observed. Midgut preparations which show strongly inhibited contractions due to previous azadirachtin trea…
Ergastoplasmic paracrystalline inclusion bodies in the adipose gonadal envelope and fat body of the glow worm, Lampyris noctiluca (Insecta, Coleoptera).
Abstract The gonads of glow worm larvae are enveloped by adipose tissue which represents a specialized fat body. The adipose gonadal envelope, and also to a lesser extent the fat body cells, contain tubular paracrystalline inclusion bodies (PIBs). Cells of other tissues are devoid of such inclusions. The PIBs form in the cisternae of rough ER. In young larvae PIB formation is sparse, but at advanced larval stages PIBs often occur as bundles in stacks of ergastoplasm. Typically, a PIB within a cisterna consists of four to seven parallel tubules. The outer diameter of a tubule is ca 28.8 nm and the width of the tubule lumen ca 12.2 nm. The “wall” of a tubule contains globular protein subunits…
The serosa of Manduca sexta (Insecta, Lepidoptera): ontogeny, secretory activity, structural changes, and functional considerations
In Manduca sexta, the blastoderm forms successively and becomes immediately cellularized as the cleavage energids reach the surface of the oocyte. Presumptive serosal cells are large and contain 2 or 4 large polyploid nuclei; presumptive embryonic cells are small and mononuclear. All parts of the blastoderm participate in the uptake and digestion of yolk material. About 10 h post-oviposition, the blastoderm breaks at the amnioserosal fold and the extraembryonic part closes above the germ band and constitutes the serosa (12 h post-oviposition, i.e. 10% development completed). At once, the serosa starts to secrete a cuticle consisting of an epi- and a lamellated endocuticle. Detachment of the…
Ultrastructure of embryonic envelopes and integument ofOncopeltus fasciatus dallas (Insecta, Heteroptera)
The embryo ofOncopeltus fasciatus forms a typical secondary dorsal organ (SDO). It develops after katatrepsis from the contracting serosa, the cells of which decrease in diameter but increase considerably in height. After 66 h, the SDO represents a protrusion of the serosal epithelium above the head and is then reduced to a disc-shaped formation, which sinks into the yolk, where it disintegrates after 80 h. During its typical expression, between 66 and 78 h, the SDO shows a zonal arrangement of its cell organelles. The nucleus, which is located in the basal cell region, has a very irregular outline and includes several nucleoli and globular inclusion bodies. Rough and smooth ER are well dev…
Studies on the fat body of oncopeltus fasciatus invaded by pseudomonas aeruginosa1
A fatal disease in laboratory cultures of Oncopeltus fasciatus is caused by Pseudomonas aeruginosa , which invades the fat body and destroys it. The cellular breakdown was studied by electron microscopy. Infected fat body tissue shrinks, disintegrates, and dissolves. The nuclei, apparently, remain intact until immediately before cell death. Bacteria, which are found singularly in cytoplasm, form an expanding electron-lucent halo. Within this halo, free ribosomes and ribosomes of rough endoplasmic reticulum (ER) disappear. The changes are presumably caused by proteolytic activity of the parasite, which results in the destruction of the fat body and death of the host. Lipid droplets dissolve,…
Ovicide-induced serosa degeneration and its impact on embryonic development in Manduca sexta (Insecta: Lepidoptera)
Abstract Eggs of Manduca sexta treated with the ovicide Ov. 165049 turn orange, and the embryos later die. The orange pigmentation is at first confined to the serosa, and is accompanied by pathological changes of serosal cells. Lipid vesicles aggregate and spindle-shaped electron-lucent vesicles—normally forming a single layer below the apical cell surface—greatly accumulate. The mitochondria swell considerably, and their matrices become electron-lucent. Subsequently, the serosal cells develop additional features of necrosis. They form many autophagic vacuoles which contain mostly degradating mitochondria, but also segregated rough endoplasmic reticulum (rER) and glycogen granules. The whol…
Electron microscopic study on the larval and adult corpus allatum of Oncopeltus fasciatus dallas (insecta, heteroptera)
1. The ultrastructure of the corpora allata of last larval instars and adults of Oncopeltus was studied. The unpaired gland undergoes submicroscopic alterations and shows signs of degradation in old animals. The organ is partly covered and penetrated by corpus cardiacum tissue. Axons with different types of neurosecretory granules form synaptoid contacts with the corpus allatum cells.
20-Hydroxyecdysone-induced differentiation and apoptosis in the Drosophila cell line, l(2)mbn
20-Hydroxyecdysone has an inhibitory effect on the proliferation of l(2)mbn cells, causes vacuolization and fragmentation of cells, and promotes a strong phagocytotic activity. From several lines of evidence, it can be concluded that 20-hydroxyecdysone induces apoptosis. Long-term video observations following the fate of individual cells, scanning and transmission electron microscopy reveal the typical characteristics of apoptosis: sequestration of small cellular protuberances or larger parts of the cell with nuclear fragments (apoptotic bodies), chromatin condensation, condensation and vesiculation of cytoplasm, whereas the mitochondria retain their normal appearance. The induction of apop…
Structural characterization and primary in vitro cell culture of locust male germline stem cells and their niche
AbstractThe establishment of in vitro culture systems to expand stem cells and to elucidate the niche/stem cell interaction is among the most sought-after culture systems of our time. To further investigate niche/stem cell interactions, we evaluated in vitro cultures of isolated intact male germline–niche complexes (i.e., apical complexes), complexes with empty niche spaces, and completely empty niches (i.e., isolated apical cells) from the testes of Locusta migratoria and the interaction of these complexes with isolated germline stem cells, spermatogonia (of transit-amplifying stages), cyst progenitor cells, cyst progenitor cell-like cells, cyst cells, and follicle envelope cells. The stru…
Differentiation and ultrastructure of the spermatogonial cyst cells in the milkweed bug,Oncopeltus fasciatus
Summary The testes of the newly hatched larva of Oncopeltus fasciatus consist of three different cell types enclosed by the tunica propria: the germline stem cells, the apical cells, and the follicle border cells which are the predecessors of the cyst progenitor cells. There are usually twelve somatically derived apical cells per follicle that no longer divide during postembryonic development. Their fine structural organization remains constant from hatching until death from old age. The apical complex cells are surrounded by at least four germline stem cells which all send cell projections toward the apical complex cells. A testicular follicle includes at least four border cells at the fir…
No evidence of androgenic hormone from the testes of the glowworm, Lampyris noctiluca.
The widely accepted concept, stating that insects have no true sex hormones, and that primary as well as secondary sex characteristics are controlled by the genetic inventory of each single cell, is challenged by the report of Naisse, J. [1966a. Controle endocrinien de la differenciation sexuelle chez l'Insecte Lampyris noctiluca (Coleoptere Malacoderme Lampyride). I. Role androgene des testicules, Arch. Biol. Liege, 77, 139-201] on the discovery of an androgenic hormone in the glowworm, Lampyris noctiluca. This case is of special interest, since it may point to an ancestral mode of sex differentiation in arthropods, considering that androgenic hormones have been discovered and characterize…
Evidence for involvement of a nuclear envelope-associated RNA helicase activity in nucleocytoplasmic RNA transport
It seems well established that translocation of at least some mRNAs through the nuclear pore is (1) an energy-dependent process, and (2) dependent on the presence of the poly(A) segment attached to most mRNA species. We describe that RNA helicase (RNA duplex unwindase) activity is present in a nuclear envelope (NE) preparation, which also appears to be involved in nucleocytoplasmic RNA transport. This activity unwinds RNA: RNA hybrids. The helicase has a pH optimum of 7.5 and a temperature optimum of 30 degrees C. Applying the sealed NE vesicle system, it was shown that duplex RNA species are readily released from the vesicles in an unidirectional manner, in contrast to single-stranded RNA,…
Differential Infectivity of Two Pseudomonas Species and the Immune Response in the Milkweed Bug, Oncopeltus fasciatus (Insecta: Hemiptera)
Pseudomonas aeruginosa and Pseudomonas putida show a profound differential infectivity after inoculation in Oncopeltus fasciatus. Whereas P. putida has no significant impact on nymphs, P. aeruginosa kills all experimental animals within 48 h. Both Pseudomonas species, however, induce the same four hemolymph peptides in O. fasciatus. Also injection of saline solution and injury induced these peptides. In general peptide induction was stronger in nymphs than in adult males. A significantly higher number of nymphs survived a challenge with P. aeruginosa when an immunization with P. putida preceded. The antibacterial properties of the hemolymph were demonstrated in inhibition experiments with P…
Midgut pseudotumors and the maintenance of tissue homeostasis: studies on aging and manipulated stick insects.
Stick insects (Carausius morosus) develop pseudotumors in aging adults. Pseudotumor formation starts at the M2 midgut region where an accumulation of stomatogastric nerve terminals is observed. Pseudotumors arise from dying columnar cells whose basal parts form an “amorphous substance” at the basement membrane whereas the apical parts, including the nucleus, are expelled into the gut lumen. The “amorphous substance” is ensheathed by hemocytes. These nodules, which do not melanize, characterize the phenotype of the pseudotumors. With age, cell death and pseudotumor infestation increases. It is shown that the maintenance of midgut tissue homoeostasis is disturbed and becomes more serious with…
Degenerative and regenerative processes involved in midgut pseudotumor formation in the stick insect (Carausius morosus)
Spontaneous and experimentally induced pseudotumor formation in Carausius morosus impairs the midgut tissue homeostasis. Spontaneous pseudotumor formation begins by the break down of a single or a small group of columnar cells (CCs) and is followed by the degeneration of neighboring CCs. There are not only marked similarities but also decisive differences between normal dying CCs in healthy specimens and the degeneration of CCs leading to pseudotumors: in both cases, the apical cell parts with the nucleus are extruded into the midgut lumen, but only during of pseudotumor formation an "amorphous substance" originates from the basal parts of the CCs. Hemocytes are attracted to this substance …
Structure and function of prothoracic glands and oenocytes in embryos and last larval instars of Oncopeltus fasciatus Dallas (Insecta, Heteroptera).
1. Active prothoracic glands and oenocytes of last larval stage are both characteristized by well-developed smooth and rough endoplasmic reticulum (ER). Prothoracic glands also show plasma membrane infoldings, but not oenocytes which contain a large number of pleomorphic vesicles. 2. The fine structure of embryonic oenocytes corresponds after blastokinesis with that of active larval and adult cells. Thus, an activity in the late embryo can be assumed. Embryonic prothoracic glands reveal no signs of activity: smooth and rough ER are absent. The subcellular structure resembles that of organ anlagen, i.e. not yet fully differentiated tissue. Hormone synthesis is not likely. 3. Ecdysone titer w…
An Incremental Analysis of the Embryonic Development of the Tobacco Hornworm,Manduca sexta
Summary A timetable of the embryonic development of Manduca sexta has been compiled to serve as a basis for physiological studies. Embryogenesis is complete within 117 hr at 24° C and was analyzed by examining specimens representing 20 stages, i.e. intervals of 5% of the total developmental time. Stage 0 denotes the newly deposited egg. Germ band formation, gastrulation, differentiation of tissues and organs, and blastokinesis are described in detail. By stage 1 (6 hr post-oviposition) the cellular blastoderm is evident and the presumptive serosal and embryonic cells are distinguishable. At stage 2 the germ band has separated from the blastoderm, and by stage 3 it has elongated enormously, …
Apoptosis of male germ-line stem cells after laser ablation of their niche.
AbstractMale germ-line stem cells (GSCs) and their niche-the apical cells or hub cells-display a unique feature at the apices of insect testicular follicles. In the locust, Locusta migratoria, the niche consists of only one large apical cell surrounded by about 60 GSCs. The apical cell can be readily identified in the intact follicle. Using laser ablation it is feasible to destroy the apical cell exclusively without injuring neighboring GSCs or any other cells. The most immediate effect on GSCs is the loss of their structural polarity. Beginning about 3 h after laser treatment chromatin starts to clump and condense in individual GSCs, and some show the first signs of cellular breakdown. The…
Tumor cell specific toxicity of Inula helenium extracts.
The aim of the research program was to identify botanical extracts with antineoplastic activity. In this respect extracts prepared from Inula helenium roots showed a remarkable activity. As evidenced by the MTT assay, the Inula helenium extract revealed a highly selective toxicity toward four different tumor cell lines (HT-29, MCF-7, Capan-2 and G1), but a much lower toxicity against healthy human peripheral blood lymphocytes (PBLs) from two donors. The extract-induced death of tumor cells was studied extensively by electron microscopy. There was a remarkable similarity of morphological alterations observed in the four cell lines: patchy chromatin condensations, cytoplasmic vesiculation, sw…
Embryonic integument and "molts" in Manduca sexta (Insecta, Lepidoptera).
In Manduca sexta the germ band is formed 12 h post-oviposition (p.o.) (=10% development completed) and is located above the yolk at the egg surface. The cells show a polar organization. They are engaged in the uptake and degradation of yolk globules, pinched off from the yolk cells. This process can be observed in the integumental cells during the first growth phase of the embryo that lasts until “katatrepsis,” an embryonic movement that takes place at 40% development completed. At 37% development completed, the ectoderm deposits a thin membrane at its apical surface, the first embryonic membrane, which detaches immediately before katatrepsis. The second period of embryonic growth—from kata…
Elektronenmikroskopische Studien �ber eine extrazellul�r auftretende Substanz in der Muskulatur des Insekts Oncopeltus fasciatus Dallas
Im Stroma von Aorta und Retrocerebralkomplex sowie von verschiedenen Muskeln (Oberschenkel-, Mitteldarm-, Speicheldrusen-, Samenleiter- und Eileitermuskeln), jedoch nicht im Stroma der Flugmuskulatur und des Herzschlauches, kommt eine fibrillar strukturierte Substanz (X-Substanz) von auserordentlich hoher Elektronendichte vor.
Expression of phase-specific haemolymph polypeptides in a laboratory strain and field catches of Schistocerca gregaria.
Uvarov's theory of locust phase polymorphism implies a differential phase-specific gene expression. It was the goal of the present investigation to provide data on the molecular level for this concept. We used the technique of 2D gel electrophoresis to generate haemolymph polypeptide maps from mature locust males. Under the given conditions we identified 238 polypeptide spots by their molecular weight and isoelectrical point. Isolated and crowded males from the Mainz strain, all originating from crowded ancesters, differed in 20 spots: three were solitary-specific and 17 crowded-specific. Field catches of solitary and gregarious S. gregaria in Mauritania revealed that those males showed the…
Ultrastructure of differentiating hemocytes in the embryo of Oncopeltus fasciatus dallas (insecta, heteroptera).
The hemocytes of Oncopeltus differentiate rather early during embryogenesis. They are segregated by the mesoderm soon after its formation (about 50h after egg deposition). Newly segregated hemocytes show the “typical” features of “embryonic” cells: many free ribosomes, a few strands of rough ER, the cisternae of which are considerably distended, electron lucent vacuoles around the periphery, and glycogen deposits. A few hours thereafter the hemocytes undergo striking subcellular changes. First, glycogen, electron lucent vacuoles and rough ER disappear and phagocytotic activity can be observed. Golgi complexes become well expressed and give rise to electron dense vesicles which fuse to large…
Case of unilateral wing formation in the female of the glowwormLampyris noctiluca
On July 27, 1999, the first author found a unilaterally winged adult glowworm in a park in the city of Mainz. Except for the wings on the left side, the specimen exhibited female characteristics that extended to external sexual appendages, the lantern and the gonads. The internal organization showed some remarkable differences between right (wingless) and left (winged) side. The right ovary contained three times more mature eggs than the left side and the volume of the corpus allatum of this side was about one-third larger than that of the left side. This suggests that aptery and egg maturation are affected by corpus allatum activity, i.e., juvenile hormone production. The findings do not s…
Binding Sites for Ca2+‐Channel Effectors and Ryanodine inPeriplaneta americana—Possible Targets for New Insecticides
The calcium channel and the 'calcium release channel' of muscle membrane of the cockroach Periplaneta americana have been characterized. Biological assays with calcium channel blockers and ryanodine on different insects and acari revealed pronounced insecticidal effects with ryanodine, but not with calcium channel blockers, at concentrations between 0.1 and 300 μg ml -1 . Skeletal muscle membranes derived either from the tubular network or from the sarcoplasmatic reticulum of P. americana were characterized with respect to the binding of the dihydropyridine (DHP) [ 3 H]isradipine (PN 200-110), the phenylalkylamine [ 3 H]verapamil and the alkaloid [ 3 H]ryanodine. Preliminary binding studies…
Die endokrinen dr�sen im embryo von Oncopeltus fasciatus Dallas (Insecta, Heteroptera)
The differentiation of the endocrine glands in the embryo of Oncopeltus fasciatus is described. The function of these glands can be correlated with the embryonic moults. The nuclei of some tissues already become polyploid in the embryo. It is discussed whether the endomitotic growth is dependent upon the function of the endocrine glands.