Increase in activity of acetylcholinesterase by 20-OH-ecdysone in a Chironomus tentans cell line

Spindler-Barth, Margarethe; Schmidt, Heinrich; Drews, Ulrich; Spindler, K.-D.

doi:10.1007/bf00375957

Cited by 21 publications

(11 citation statements)

References 23 publications

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“…There have been several reports on the stimulation of AchE activity by molting hormones in various insect cell lines (Cherbas et al, 1977; Archives of Insect Biochemistry and Physiology et al., 1978;Berger and Wyss, 1980;Cohen, 1981;Spindler et al, 1988;Spindler-Barth, 1991). However, the physiological significance of these results remains unknown.…”

Section: Discussionmentioning

confidence: 95%

“…Most convincing evidence for non-neuronal AchE activity was the appearance of AchE during vertebrate morphogenesis, as a part of a muscarinic cholinergic system that may be involved in cell movement during differentiation (Oettling et al, 1985). In addition, a correlation was observed between the induction of morphological changes by molting hormones and the increase in specific AchE activity in the C. tentans cell line (Spindler-Barth et al, 1988). The induction of AchE by ecdysteroids was a successful, sensitive, and simple test for the detection of molting hormone agonists.…”

Section: Discussionmentioning

confidence: 95%

“…Cherbas et al (1977) suggested that the presence and stimulation of this enzyme by 20E may indicate that the cells are of neuronal origin. Nevertheless, the basal level of AchE activity in the Drosophila cell line was only 2% of the AchE activity in Drosophila nervous tissue (Spindler-Barth et al, 1988). In addition, Berger and Wyss (1980) demonstrated by electrophoresis that the AchE from Drosophila brain differed from the one expressed in the S3 Drosophila cell line.…”

Section: Discussionmentioning

confidence: 98%

“…The change in absorbance at 412 nm was recorded during 10 min using the kinetic mode of a 96-well microtiter plate spectrophotometer (MultiPower wave X-340) and the initial rate was estimated. The true AchE activity was defined as the net rate of eserine-sensitive hydrolysis of acetylthiocholine iodide, which means the overall rate of hydrolysis minus any component not eliminated by 10 µM eserine (Spindler-Barth et al, 1988).…”

Section: Archives Of Insect Biochemistry and Physiologymentioning

confidence: 99%

“…The cessation of cell growth, observed as a decrease in cell proliferation, is also indicative of an ecdysteroid action. In addition, induction of acetylcholinesterase (AchE) and competition with 3 H-ponasteroneA (PoA) for EcR binding can be used to prove an ecdysteroid activity (Spindler-Barth et al, 1988).…”

Section: Introductionmentioning

confidence: 99%

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Action of 24-epibrassinolide on a cell line of the beet armyworm,Spodoptera exigua

Decombel

Tirry

Smagghe

2005

Arch. Insect Biochem. Physiol.

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The Spodoptera exigua cell line Se4 is sensitive for ecdysteroid activity stimulated by the insect molting hormone, 20-hydroxyecdysone (20E), showing a cease in cell proliferation (with 50% inhibition around 1 microM) and characteristic cell morphology changes with aggregation and formation of long filamentous cytoplasmic extensions. The bisacylhydrazine tebufenozide also triggered such typical cellular effects in Se4, and in addition, it showed an affinity for binding in competition with 3H-ponasterone A (PoA) that was similar to 20E (with 50% competition around 1 microM), confirming that such non-ecdysteroids display an ecdysteroid agonist activity. In contrast, when Se4 cells were incubated with the native plant hormone 24-epibrassinolide (24BR), none of the effects triggered by 20E were observed. Hence, a competition binding experiment with 3H-PoA demonstrated no affinity of 24BR for binding to the ecdysteroid receptor in the Se4 cell line. In another series of experiments, the Se4 cell line was tested in sensitivity response to increased acetylcholinesterase (AchE) activity after treatment with ecdysteroid active compounds. The AchE activity measured in the cell line is discussed in relation to inhibition by eserine. The obtained results suggest that 24BR exerted no ecdysteroid activity.

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Section: Discussionmentioning

confidence: 95%

Section: Discussionmentioning

confidence: 95%

Section: Discussionmentioning

confidence: 98%

Section: Archives Of Insect Biochemistry and Physiologymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Action of 24-epibrassinolide on a cell line of the beet armyworm,Spodoptera exigua

Decombel

Tirry

Smagghe

2005

Arch. Insect Biochem. Physiol.

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Presence of Muscarinic Acetylcholine Receptors in the Cattle TickBoophilus microplusand in Epithelial Tissue Culture Cells ofChironomus tentans

et al. 1996

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A muscarinic acetylcholine receptor (mAChR) has been demonstrated and partially characterized in larvae of the cattle tick Boophilus microplus. Its properties are compared with mAChR from an epithelial cell line from the dipteran insect Chironomus tentans. Competition studies with cholinergic ligands of different specificity revealed the muscarinic nature of the cholinergic receptors investigated in both species. In homogenates from tick larvae, specific binding sites for [3H]quinuclidinyl benzilate (QNB) with high affinity (1·2±(0·13) nM; Bmax 22·5 pmol mg protein−1) were detected that do not bind nicotinic compounds specifically. The estimated IC50 values for nicotine, imidacloprid and α‐bungarotoxin were all in the mM range. Additionally, with tick larvae, high‐affinity nicotinic binding sites were detected with [3H]nicotine which could be displaced by high concentrations of imidacloprid or QNB. The estimated IC50 values for nicotine, α‐bungarotoxin, imidacloprid and QNB were 43(±8) nM, 0·8(±0·2) μM, 2·8(±0·6) μM and 78(±1·9) μM, respectively. With homogenates of the non‐neuronal insect cell line from C. tentans, only high‐affinity binding sites for [3H]QNB were found. Muscarinic antagonists selectively displaced [3H]quinuclidinyl benzilate (QNB) binding to tick larvae homogenates. The mAChR of B. microplus preferred pirenzepine (IC50 2·13(±1·02) μM) among different subtype‐specific mAChR antagonists (4‐DAMP had IC50 49·9(±9·13) μM and methoctramine had IC50 121(±14·2) μM) indicating a type of binding site similar to the vertebrate M1 mAChR subtype. The tick muscarinic receptor seems to be a G‐protein‐coupled receptor, as concluded from the 4·8‐fold reduction in receptor affinity for binding of the muscarinic agonist oxotremorine M upon treatment with the non‐hydrolysable GTP‐analogue γ‐S‐GTP. Binding data for the agonists oxotremorine M (IC50 71·3(±19·6) μM) and carbachol (IC50 253(±87·1) μM) parallel the biological efficacy of these compounds, in that, while oxotremorine M showed some activity against ticks, carbachol was ineffective.

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On the action of RH 5849, a nonsteroidal ecdysteroid agonist, on a cell line from Chironomus tentans

Spindler-Barth

Turberg

Spindler

1991

Arch Insect Biochem Physiol

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RH 5849 competes with 3H-ponasterone A for ecdysteroid binding sites in Chironomus tentans cells with an about fourfold lower relative affinity as compared to 20-OH-ecydsone. It does not interfere with glucocorticoid and estrogen binding sites in vertebrates. It i s also not recognized by an ecdysteroid antibody. RH 5849 exerts typical morphological and physiological effects ascribed to the action of 20-OH-ecdysone o n C. tentans cells, namely an increase in acetylcholinesterase activity and an inhibition of chitin synthesis.

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Increase in activity of acetylcholinesterase by 20-OH-ecdysone in a Chironomus tentans cell line

Cited by 21 publications

References 23 publications

Action of 24-epibrassinolide on a cell line of the beet armyworm,Spodoptera exigua

Action of 24-epibrassinolide on a cell line of the beet armyworm,Spodoptera exigua

Presence of Muscarinic Acetylcholine Receptors in the Cattle TickBoophilus microplusand in Epithelial Tissue Culture Cells ofChironomus tentans

On the action of RH 5849, a nonsteroidal ecdysteroid agonist, on a cell line from Chironomus tentans

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