The in vitro development of the pupal integument and the effects of ecdysteroids in Tenebrio molitor (Insecta, Coleoptera)

Quennedey, André; Quennedey, Brigitte; Delbecque, Jean‐Paul; Delachambre, Jean

doi:10.1007/bf00216424

Cited by 32 publications

(7 citation statements)

References 44 publications

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“…Forewings from pupae taken at different ages were dissected, the pupal cuticle removed, the upper wing epidermis isolated and placed into a ninety‐six‐well plate containing Landureau's S20 medium (Landureau & Grellet, 1972), modified according to the Na + /K + ratio (65 : 42) and osmolality (380 mOsm) of Tenebrio haemolymph (Quennedey et al ., 1983), supplemented with 5% fetal bovine serum (FBS) (Seromed) and, according to the experiments, 0.1–100 µ m 20E (Sigma) dissolved in ethanol. Plates were incubated at 25 °C 1 day before transfection.…”

Section: Methodsmentioning

confidence: 99%

A functional analysis of ACP‐20, an adult‐specific cuticular protein gene from the beetle Tenebrio: role of an intronic sequence in transcriptional activation during the late metamorphic period

Lemoine¹,

Mathelin²,

Braquart‐Varnier³

et al. 2004

Insect Molecular Biology

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A gene encoding the adult cuticular protein ACP-20 was isolated in Tenebrio. It consists of three exons interspersed by two introns, intron 1 interrupting the signal peptide. To understand the regulatory mechanisms of ACP-20 expression, ACP-20 promoter-luciferase reporter gene constructs were transfected into cultured pharate adult wing epidermis. Transfection assays needed the presence of 20-hydroxyecdysone, confirming that ACP-20 is up-regulated by ecdysteroids. Analysis of 5' deletion constructs revealed that three regions are necessary for high levels of transcription. Interaction experiments between intronic fragments and epidermal nuclear proteins confirmed the importance of intron 1 in ACP-20 transcriptional control, which results from the combined activity of regulatory cis-acting elements of the promoter and those of intron 1.

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Section: Methodsmentioning

confidence: 99%

A functional analysis of ACP‐20, an adult‐specific cuticular protein gene from the beetle Tenebrio: role of an intronic sequence in transcriptional activation during the late metamorphic period

Lemoine¹,

Mathelin²,

Braquart‐Varnier³

et al. 2004

Insect Molecular Biology

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“…It is also important to note, however, that pupal procuticle deposition occurs even in the continuous presence of ecdysone in Drosophila (references in Fristrom et al, 1982) see for example (Fristrom et al, 1973;Johnson and Milner, 1990;Milner and Muir, 1987), yet at slower rates than under a pulse regimen of hormone. Similarly, adult sternal cuticle of Tenebrio was synthesized in vitro in the continuous presence of peak levels (4 mg/ml) of 20E, but at a much slower rate than in vivo (Quennedey et al, 1983). Culture parameters may be also important, as for procuticle deposition in vitro, the inhibitory effect of 20E is not as important in the presence of BSA in the culture medium (Fristrom and Yund, 1980).…”

Section: Importance Of the Ecdysteroid Pulsementioning

confidence: 97%

The regulation of expression of insect cuticle protein genes

Charles

2010

Insect Biochemistry and Molecular Biology

150

102

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“…The specific role of ecdysone, in vivo and in vitro, on epidermal differentiation has been observed in Schistocerca gregaria and Tenebrio molitor (Quennedey et al 1983). Locust epidermis cultured in a hormone-free medium or in a medium containing ecdysone detaches from the overlying cuticle (pseudo-apolysis).…”

Section: Discussionmentioning

confidence: 96%

Ultrastructural studies of the in vitro chitin and cuticle secretions in the wing pads of Locusta

Cassier¹,

Papillon²

1991

Can. J. Zool.

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CASSIER, P., and PAPILLON, M. 1991. Ultrastructural studies of the in vitro chitin and cuticle secretions in the wing pads of Locusta. Can. J . Zool. 69: [29][30][31][32][33][34][35][36][37][38].Cuticle deposition and chitin synthesis in the wing pads of 1-day-old fifth-instar larvae of Locusta were stimulated in vitro by 20-hydroxyecdysone and makisterone A. Stimulation was dose-dependent from 2 X lop7 to 2 X lop5 M. The response appeared after a 36-to 48-h lag period characterized by inhibition of the postecdysial activity of the previous cuticular cycle. The hormones stimulated deposition of the epicuticular layers, triggered uptake of the chitin precursor N-acetyl-D-glucosamine and biosynthesis of chitin, and initiated microtubular bundle orientation. Ecdysone has no effect on these processes. Ecdysone and 20-hydroxyecdysone do not act synergistically on chitin biosynthesis. These data confirm earlier observations on the differential effects of the major forms of ecdysteroids, ecdysone and 20-hydroxyecdysone, on the activity of epidermal cells of locusts. Chitin biosynthesis by 20-hydroxyecdysone stimulated cells requires the integrity of the microtubular system and the chitin-synthetase complex. It was inhibited by colcemid, a microtubular poison, and by diflubenzuron, an inhibitor of chitin synthetase. These drugs had no effect on apolysis and epicuticular secretion. The in vitro biosynthesis of chitin was also inhibited by azadirachtin, a cytotoxic drug. CASSIER, P., et PAPILLON, M. 1991. Ultrastructural studies of the in vitro chitin and cuticle secretions in the wing pads of Locusta. Can. J . Zool. 69 : 29-38.Le dCpBt de la cuticule et la biosynthese de la chitine par les ptkrotheques de larves du cinquieme stade de Locusta, BgCes de 1 jour, sont stimulCs in vitro par la 20-hydroxyecdysone et par la makistkrone A. La stimulation est fonction de la dose aux concentrations comprises entre 2 x lop7 et 2 X M. La rCponse apparait apres une pCriode de latence de 36-48 h caractCrisCe par l'inhibition Qe I'activitC post-exuviale du cycle cuticulaire prickdent. Les hormones favorisent le dCpBt des assises Cpicuticulaires, stimulent l'incorporation de la N-acCty1-D-glucosamine, prCcurseur de la chitine, dCclenchent la biosynthese de la chitine et la disposition en faisceaux des microtubules. L'ecdysone n'a pas d'effet significatif sur ces processus; en revanche, elle induit la diffkrenciation cellulaire. L'ecdysone et la 20-hydroxyecdysone n'ont pas d'effet synergique sur la biosynthese de la chitine. Ces observations confirment les donnCes antirieures quant h l'action diffkrentielle des ecdystkroi'des majeurs, l'ecdysone et la 20-hydroxyecdysone, sur I'activitC des cellules Cpidermiques des Criquets. La biosynthese de la chitine par les cellules stimulkes par la 20-hydroxyecdysone requiert I'intCgritC du systeme microtubulaire et de la chitine-synthCtase. Elle est inhibCe par la colcCmide, poison microtubulaire, et par le diflubenzuron, inhibiteur de la chitine-synthCtase. Ces drogues n'ont pas d'effet sur l'apol...

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The in vitro development of the pupal integument and the effects of ecdysteroids in Tenebrio molitor (Insecta, Coleoptera)

Cited by 32 publications

References 44 publications

A functional analysis of ACP‐20, an adult‐specific cuticular protein gene from the beetle Tenebrio: role of an intronic sequence in transcriptional activation during the late metamorphic period

A functional analysis of ACP‐20, an adult‐specific cuticular protein gene from the beetle Tenebrio: role of an intronic sequence in transcriptional activation during the late metamorphic period

The regulation of expression of insect cuticle protein genes

Ultrastructural studies of the in vitro chitin and cuticle secretions in the wing pads of Locusta

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