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Biomedical subjects

R Lanot

Publications and source records attributed to R Lanot.

At least 19 recordsLinked to original sources

Postembryonic hematopoiesis in Drosophila.

We have investigated the blood cell types present in Drosophila at postembryonic stages and have analysed their modifications during development and under immune conditions. The anterior lobes of the larval hematopoietic organ or lymph gland contain numerous active secretory cells, plasmatocytes, few crystal cells, and a number of undifferentiated prohemocytes. The posterior lobes contain essentially prohemocytes. The blood cell population in larval hemolymph differs and consists mainly of plasmatocytes which are phagocytes, and of a low percentage of crystal cells which reportedly play a role in humoral melanisation. We show that the cells in the lymph gland can differentiate into a given blood cell lineage when solicited. Under normal nonimmune conditions, we observe a massive differentiation into active macrophages at the onset of metamorphosis in all lobes. Simultaneously, circulating plasmatocytes modify their adhesion and phagocytic properties to become pupal macrophages. All phagocytic cells participate in metamorphosis by ingesting doomed larval tissues. The most dramatic effect on larval hematopoiesis was observed following infestation by a parasitoid wasp. Cells within all lymph gland lobes, including prohemocytes from posterior lobes, massively differentiate into a new cell type specifically devoted to encapsulation, the lamellocyte.

Animals↗

Insect immunity. Constitutive expression of a cysteine-rich antifungal and a linear antibacterial peptide in a termite insect.

Two novel antimicrobial peptides, which we propose to name termicin and spinigerin, have been isolated from the fungus-growing termite Pseudacanthotermes spiniger (heterometabole insect, Isoptera). Termicin is a 36-amino acid residue antifungal peptide, with six cysteines arranged in a disulfide array similar to that of insect defensins. In contrast to most insect defensins, termicin is C-terminally amidated. Spinigerin consists of 25 amino acids and is devoid of cysteines. It is active against bacteria and fungi. Termicin and spinigerin show no obvious sequence similarities with other peptides. Termicin is constitutively present in hemocyte granules and in salivary glands. The presence of termicin and spinigerin in unchallenged termites contrasts with observations in evolutionary recent insects or insects undergoing complete metamorphosis, in which antimicrobial peptides are induced in the fat body and released into the hemolymph after septic injury.

Amino Acid Sequence↗

Treatment of l(2)mbn Drosophila tumorous blood cells with the steroid hormone ecdysone amplifies the inducibility of antimicrobial peptide gene expression.

Insects rely on both humoral and cellular mechanisms to defend themselves against microbial infections. The humoral response involves synthesis of a battery of potent antimicrobial peptides by the fat body and, to a lesser extent, by blood cells. The cellular response on the other hand consists of phagocytosis of small microorganisms and melanization and encapsulation of larger parasites. The l(2)mbn cell line, established from tumorous larval hemocytes, represents a system of choice to dissect the molecular events controlling cellular immunity. We report here that l(2)mbn cells can be efficiently induced to differentiate in adherent, macrophage-like cells by treatment with 20-hydroxyecdysone. Ecdysone treatment increases both the phagocytic capacity of l(2)mbn cells and their competence to express antimicrobial genes in response to immune challenge. We also report that expression of several regulatory molecules thought to be involved in the immune response is up-regulated by ecdysone in l(2)mbn cells.

Animals↗

Drosophila immunity: analysis of larval hemocytes by P-element-mediated enhancer trap.

Our aim was to identify new genes involved in the cellular aspects of defense mechanism of Drosophila, as well as in melanotic tumor formation processes that are linked to blood cell disregulation. We have screened 1341 enhancer detector fly lines for expression of the lacZ reporter gene in larval hemocytes at the end of the third instar. We have selected 21 lines in which we observed a reproducible lacZ expression in blood cells. These lines were classified according to the subsets of hemocytes in which lacZ was expressed, and we identified five lines that can be used as lamellocyte markers. Three lines were selected for further analysis. The first exhibited strong lacZ expression in all lamellocytes. The second expressed lacZ in plasmatocytes and lamellocytes, and exhibited a melanotic tumor phenotype in larvae homozygous for the insertion. A third line showed a striking insertion-linked phenotype of melanized lymph glands (the hematopoietic organ), which resulted in the total absence of circulating hemocytes in the mutant larvae. We anticipate that this mutation, which we named domino, will prove a useful tool in the analysis of the role of hemocytes during the various aspects of immune response and melanotic tumor formation.

Animals↗

Novel inducible antibacterial peptides from a hemipteran insect, the sap-sucking bug Pyrrhocoris apterus.

Insects belonging to the recent orders of the endopterygote clade (Lepidoptera, Diptera, Hymenoptera and Coleoptera) respond to bacterial challenge by the rapid and transient synthesis of a battery of potent antibacterial peptides which are secreted into their haemolymph. Here we present the first report on inducible antibacterial molecules in the sap-sucking bug Pyrrhocoris apterus, a representative species of the Hemiptera, which predated the Endoptergotes by at least 50 million years in evolution. We have isolated and characterized from immune blood of this species three novel peptides or polypeptides: (i) a 43-residue cysteine-rich anti-(Gram-positive bacteria) peptide which is a new member of the family of insect defensins; (ii) a 20-residue proline-rich peptide carrying an O-glycosylated substitution (N-acetylgalactosamine), active against Gram-negative bacteria; (iii) a 133-residue glycine-rich polypeptide also active against Gram-negative bacteria. The proline-rich peptide shows high sequence similarities with drosocin, an O-glycosylated antibacterial peptide from Drosophila, and also with the N-terminal domain of diptericin, an inducible 9 kDa antibacterial peptide from members of the order Diptera, whereas the glycine-rich peptide has similarities with the glycine-rich domain of diptericin. We discuss the evolutionary aspects of these findings.

Amino Acid Sequence↗

Characterization and transcriptional profiles of a Drosophila gene encoding an insect defensin. A study in insect immunity.

Insect defensins are a family of 4-kDa, cationic, inducible antibacterial peptides which bear six cysteine residues engaged in three intramolecular disulfide bridges. They owe their name to certain sequence similarities with defensins from mammalian neutrophiles and macrophages. We report the characterization of a novel defensin isoform from Drosophila and the cloning of the gene encoding a preprodefensin. The gene, which is intronless and present in a single copy/haploid genome, maps at position 46CD on the right arm of the second chromosome. The analysis of the upstream region of the gene reveals the presence of multiple putative cis-regulatory sequences similar to mammalian regulatory motifs of acute-phase-response genes. Transcriptional profiles indicate that the Drosophila defensin gene is induced by bacterial challenge with acute-phase kinetics. It is also expressed in the absence of immune challenge during metamorphosis. These and other data on the Drosophila defensin gene lead us to suggest that insect and mammalian defensins have evolved independently.

Amino Acid Sequence↗

Effects of an imidazole derivative (KK-42) on development and ecdysteroid production in Locusta migratoria (Insecta, Orthoptera).

The imidazole derivative KK-42 was applied in various experimental conditions to larvae and adult females of Locusta migratoria. The effect of this compound was monitored on the development of larvae, on oocyte growth in adult females and on the development of eggs laid by these females. KK-42 had only minor effects on postembryonic development; anticipation of imaginal moult was never observed. In contrast oocyte and egg development were markedly affected by KK-42: this effect is however not related to modifications of the synthesis of ecdysteroids in the ovaries.

Animals↗

Further experimental evidence for the involvement of ecdysone in the control of meiotic reinitiation in oocytes of Locusta migratoria (Insecta, Orthoptera).

Ecdysone has recently been shown to be able to trigger meiotic reinitiation in vitro in submature oocytes of Locusta. In the present study we have experimentally depressed (by 60-70%) ecdysone biosynthesis in the ovaries of adult females by rearing them on a diet with a modified sterol profile. Mature oocytes from such females fail to undergo normal reinitiation, but when incubated in vitro, can be induced to break their meiotic arrest by the addition of exogenous ecdysone. These results lend further support to the hypothesis that in Locusta, ovarian ecdysone is involved in the control of meiotic reinitiation.

Animals↗

Ecdysteroid biosynthesis and embryonic development are disturbed in insects (Locusta migratoria) reared on plant diet (Triticum sativum) with a selectively modified sterol profile.

Wheat seedlings germinating in the presence of the systemic fungicide fenpropimorph accumulate 9beta,19-cyclopropylsterols (95% of total sterols) in place of Delta(5)-sterols, which are normally produced in these plants. Adult females of the phytophagous insect Locusta migratoria show a dramatic decrease in their cholesterol content when reared on fenpropimorph-treated wheat. These females lay eggs with the ecdysteroid concentration reduced by up to 80% as compared to controls. Injection of fenpropimorph to the insects or feeding them on wheat coated with the fungicide (normal sterol composition) does not affect their sterol or ecdysteroid profiles; addition of cholesterol to fenpropimorph-treated wheat prior to feeding restores normal ecdysteroid titers in the insects. The severe reduction of the ecdysteroid content in eggs laid by females reared on fenpropimorph-treated wheat is associated with a series of developmental arrests and/or abnormalities. The results show that the dietary 9beta,19-cyclopropylsterols cannot be used by Locusta in place of Delta(5)-sterols for ecdysteroid biosynthesis. They suggest that the selective inhibition of specific enzymes in the sterol biosynthetic pathway of the plants can be used as a strategy to control insect development.

Journal Article↗

The larval integument of Calliphora erythrocephala Meigen (Diptera : Calliphoridae) during metamorphosis. Changes in macromolecular content and in incorporation rate of labelled precursors.

The amount of proteins, RNA and DNA in the larval integument of Calliphora erythrocephala, decreased markedly when metamorphosis began. The total incorporation per epidermis of labelled precursors also decreased at this time, but far less in RNA than in DNA and proteins. Thus, incorporation, when expressed in terms of specific activity, decreased in proteins, remained stable in DNA and increased in RNA. The significance of these results is discussed.

Animals↗

Conversion of a radiolabelled ecdysone precursor, 2,22,25-trideoxyecdysone, by embryonic and larval tissues of Locusta migratoria.

A high specific activity tritiated ecdysone precursor, 2,22,25-trideoxyecdysone, was used to probe the capacity of various embryonic and larval tissues to perform the last 3 hydroxylation steps in ecdysone biosynthesis. Embryos at early stages of development, prior to the differentiation of their endocrine glands and embryonic heads, thoraces and abdomens of later stages, were found to have the capacity to hydroxylate the precursor to ecdysone. Larval epidermis and fat body are also able to transform 2,22,25-trideoxyecdysone into ecdysone; Malpighian tubules and midgut hydroxylate the precursor at C-2 but are apparently unable to hydroxylate both at C-22 and C-25. Larval prothoracic glands convert the precursor to ecdysone at a very efficient rate, which is 1-2 magnitudes higher than that of the other tissues investigated; several data argue for the existence of a privileged sequence of hydroxylations, C-25, C-22, C-2, in the larval prothoracic glands.

Abdomen↗

[Regression of chick embryo tail gut. Stage of determination of cell degeneration and role of the associated mesenchyme].

In chick embryo the complete degeneration of the tail gut is carried out between H and H stages 20 to 24 and cell alterations are noticeable from stage 19. This paper reports a study to ascertain whether degeneration of the tail gut cells is already determined. The tail guts of donor embryos were taken at stages 17 to 20 and implanted into the leg bud of 4 to 5-day host embryos. In another series, stage 19 tail guts were implanted with their associated mesenchyme. The fate of the implants was histologically studied in the hosts at different times during their subsequent development. The younger grafts formed tubular structures almost identical to the normal tail gut. Implanted stage 19 tail guts with practically no mesenchyme did not degenerate; they maintained their structure but did not differentiate further. The tail guts implanted with associated mesenchyme degenerated, and stage 20 tail guts also regressed. The death of the tail gut cells was not determined until the degeneration processes had actually begun, the fate of the tail gut cells depending on their environment.

Animals↗

[Proteins, nucleic acids and acid hydrolases in the chick embryo tail bud].

During the 3d and 4th days of incubation the tail bud displays cell degeneration which result in the complete involution of both postanal gut and primitive streak. In spite of the degenerations the amounts of DNA, RNA and proteins are still growing. Over this period the activities of acid phosphatase and cathepsin are changing. The specific activity of phosphatase reaches a maximum when the tissue resorption is most intensive. The specific activity of cathepsin is then at its lowest level but it has passed earlier through a maximum.

Acid Phosphatase↗

[Tail gut regression and associated phenomena in chick embryo (author's transl)].

The regression of the tail gut and the associated phenomena in chick embryo were studied using cytological and biochemical methods. The cells of the tail gut begin degenerating at stage 20. The regression is completed at stage 24. The remnant of the primitive streak which have been incorporated into the tail bud degenerate at the same time. During this period dead cells are sporadically found in the other tissues of the tail bud, chiefly in the mesenchym. The morphological characteristics of cell degeneration and elimination, in the tail gut as in the mesenchyme, are defined. The biochemical study showed that the degenerative phenomena correspond to an enhanced activity of acid phosphatase. This study shows further that these phenomena do not change the total growth rate of the tail bud while they are developing.

Acid Phosphatase↗

[Tooth regeneration in the guinea pig (Cavia porcellus)].

The first inferior molar has been extracted, a part of its being reimplanted or not. A new molar of normal form regenerated, apparently from the apex of the tooth germ, in all cases in which the alveolus was left free or implanted with a tooth freagment deprived of pulpa.

Animals↗