Patterns of juvenile hormone have been intensively studied in the cockroach Blattella germanica under different physiological situations. However, data have been mainly obtained in vitro, and refer ...to hormone synthesized by isolated corpora allata, whereas information available on hormone concentration in the hemolymph is restricted to adult females. In order to complement our studies in vitro, we have measured juvenile hormone titer in the hemolymph of B. germanica females in four characteristic physiological situations: penultimate and last instar nymphs, adults during the first vitellogenic cycle, and adults transporting egg cases (ootheca). In general, a significant positive correlation between rates of hormone synthesis and concentration in the hemolymph is observed. The main disparities appear in the penultimate day of the period of ootheca transport, where titer is high whereas synthesis is low, and on day 6 of the first vitellogenic cycle, where synthesis increases whereas titer decreases. At these stages, the observed disparities between synthesis and titer might be explained by differential action of degradation enzymes.
Une recherche sur les noctuelles nous a permis d'elever des larves de nombreuses especes de lepidopteres, principalement des noctuelles, sur un substrat artificiel du nom de <<McMorran diet>> en ...laboratoire. Nous reportons le developpement de 103 especes de lepidopteres, dont 39 especes qui n'ont pas encore ete documentees, comme pouvant se developper sur ce substrat artificiel. Etant donne son faible cout, facilite de preparation, et large champ d'action, ce substrat artificiel peut grandement faciliter la recherche sur les lepidopteres et autres insectes en laboratoire.
New cell lines from embryos of Ephestia kuehniella were recently developed. Primary cultures were initiated in September 2002 from 2 to 4 day old eggs in either modified TC-100 or ExCell 400 medium. ...From these initial cultures, one, originally isolated in the Ex-Cell medium, produced sufficient cell growth to allow subcultivation and eventually led to the establishment of two cell strains, one that forms multicellular vesicles in suspension and one consisting of tightly attached epithelial-like cells. The strains were compared to an extract from E. kuehniella eggs by isozyme analysis and shown to be from the same species. Both strains were inoculated with various insect viruses, including nucleopolyhedroviruses from Autographa californica, Anagrapha falcifera, Anticarsa gemmatalis, Galleria mellonella, Heliothis armigera, Helicoverpa zea, Lymantria dispar, Plutella xylostella, and Rachoplusia ou. Both strains were highly susceptible to most of the nucleopolyhedroviruses (with the exception of the viruses from Helicoverpa zea and Lymantria dispar which did not show cytopathology to either cell strain) with large numbers of occlusion bodies produced in most of the inoculated cells. Our results suggest these new lines can be useful in biocontrol research. NPV nucleopolyhedrovirus, the prefix M or S refers to multiple or single nucleocapsids Ek-x4T an attached cell line from E. kuehniella Ek-x4V a vesicular cell line from E. kuehniella nucleopolyhedrovirus, the prefix M or S refers to multiple or single nucleocapsids an attached cell line from E. kuehniella a vesicular cell line from E. kuehniella
Topical application of 400µg of the juvenile hormone analog, methoprene, to females of the penultimate instar of Leucophaea maderae failed to induce vitellogenin synthesis. However, last instar ...females showed an increasing response level in making vitellogenin as they aged during the first half of the instar. In the second half of the last instar the response to methoprene declined to nearly zero when the prothoracic glands have become highly active. Then, a few days before the metamorphic molt the responsiveness reached maximal levels, i.e., comparable to adult females. These data suggest that the fat body develops competency to produce vitellogenin during the last nymphal instar, but increasing titers of ecdysone then interfere with the action of methoprene and consequently production of vitellogenin is curtailed. When prothoracic glands from the second half of the last instar were implanted into adult females, the normal activation of the corpora allata, or their accelerated activation induced by mating, did not occur. Likewise, an activation of the corpora allata due to the severance of the NCCI was not observed when prothoracic glands had been implanted prior to such operations. Thus, ecdysone released by the prothoracic glands appeared to directly inhibit the isolated corpora allata in vivo i.e. without the mediation by the brain. Methoprene applied to allatectomized adult females induced vitellogenin synthesis in a dose dependent manner. This induction was, however, quantitatively reduced by implanted active prothoracic glands, particularly when low doses of methoprene had been applied. Methoprene higher than 5µg overcame the inhibitory potency of the implanted prothoracic glands. The effect of the prothoracic glands, i.e. ecdysone, appears to signal an interference with the action of methoprene at the target tissues, the fat body. The exposure of the fat body to a given juvenile hormone/ecdysone ratio dictates the apparent effectiveness of ecdysone. The precise mode of the interaction of juvenile hormone and ecdysone on the adult fat body is not known. These data show that ecdysone inhibits vitellogenesis by two independent mechanisms: 1) inhibition of the corpora allata resulting in the inhibition of juvenile hormone production and 2) inhibition of vitellogenin synthesis by the fat body. Both of these mechanisms appear to be operative in immature and mature animals. However, the action of ecdysones on the fat body is only seen after it had acquired competency to make vitellogenin during the last nymphal instar.
New cell lines from embryos of Ephestia kuehniella were recently developed. Primary cultures were initiated in September 2002 from 2 to 4 day old eggs in either modified TC-100 or ExCell 400 medium. ...From these initial cultures, one, originally isolated in the Ex-Cell medium, produced sufficient cell growth to allow subcultivation and eventually led to the establishment of two cell strains, one that forms multicellular vesicles in suspension and one consisting of tightly attached epithelial-like cells. The strains were compared to an extract from E. kuehniella eggs by isozyme analysis and shown to be from the same species. Both strains were inoculated with various insect viruses, including nucleopolyhedroviruses from Autographa californica, Anagrapha falcifera, Anticarsa gemmatalis, Galleria mellonella, Heliothis armigera, Helicoverpa zea, Lymantria dispar, Plutella xylostella, and Rachoplusia ou. Both strains were highly susceptible to most of the nucleopolyhedroviruses (with the exception of the viruses from Helicoverpa zea and Lymantria dispar which did not show cytopathology to either cell strain) with large numbers of occlusion bodies produced in most of the inoculated cells. Our results suggest these new lines can be useful in biocontrol research.
Three ecdysteroid 7,9(11)-dien-7-ones (dacryhainansterone, 25-hydroxydacryhainansterone and kaladasterone) were prepared by dehydration of the corresponding 11a-hydroxy ecdysteroids (ajugasterone C, ...turkesterone and muristerone A, respectively). The biological activities of the dienones in the Drosophila melanogaster B(II) cell bioassay, which reflect the affinity for the ecdysteroid receptor complex, showed that the dienones retain high biological activity. Irradiation at 350 nm of the ecdysteroid dienones (100 nM) with bacterially-expressed dipteran and lepidopteran ecdysteroid receptor proteins (DmEcR/DmUSP or CfEcR/CfUSP), followed by loading with (3)Hponasterone A revealed that irradiation of dacryhainansterone or kaladasterone resulted in blocking of >70% of the specific binding sites. Thus, ecdysteroid dienones show considerable potential as photoaffinity analogues for ecdysteroid binding proteins.
Topical application of 400 micro g of the juvenile hormone analog, methoprene, to females of the penultimate instar of Leucophaea maderae failed to induce vitellogenin synthesis. However, last instar ...females showed an increasing response level in making vitellogenin as they aged during the first half of the instar. In the second half of the last instar the response to methoprene declined to nearly zero when the prothoracic glands have become highly active. Then, a few days before the metamorphic molt the responsiveness reached maximal levels, i.e., comparable to adult females. These data suggest that the fat body develops competency to produce vitellogenin during the last nymphal instar, but increasing titers of ecdysone then interfere with the action of methoprene and consequently production of vitellogenin is curtailed. When prothoracic glands from the second half of the last instar were implanted into adult females, the normal activation of the corpora allata, or their accelerated activation induced by mating, did not occur. Likewise, an activation of the corpora allata due to the severance of the NCCI was not observed when prothoracic glands had been implanted prior to such operations. Thus, ecdysone released by the prothoracic glands appeared to directly inhibit the isolated corpora allata in vivo i.e. without the mediation by the brain. Methoprene applied to allatectomized adult females induced vitellogenin synthesis in a dose dependent manner. This induction was, however, quantitatively reduced by implanted active prothoracic glands, particularly when low doses of methoprene had been applied. Methoprene higher than 5 micro g overcame the inhibitory potency of the implanted prothoracic glands. The effect of the prothoracic glands, i.e. ecdysone, appears to signal an interference with the action of methoprene at the target tissues, the fat body. The exposure of the fat body to a given juvenile hormone/ecdysone ratio dictates the apparent effectiveness of ecdysone. The precise mode of the interaction of juvenile hormone and ecdysone on the adult fat body is not known. These data show that ecdysone inhibits vitellogenesis by two independent mechanisms: 1) inhibition of the corpora allata resulting in the inhibition of juvenile hormone production and 2) inhibition of vitellogenin synthesis by the fat body. Both of these mechanisms appear to be operative in immature and mature animals. However, the action of ecdysones on the fat body is only seen after it had acquired competency to make vitellogenin during the last nymphal instar.