Cytochalasin D suppresses beta‐adrenergic induced protein discharge without inhibiting membrane fusion

Busson-Mabillot, S; Chambaut-Guérin, Anne-Marie; Huleux-Maurs, C.; Ovtracht, L; Rossignol, B.

doi:10.1111/j.1768-322x.1985.tb00367.x

Cited by 6 publications

(11 citation statements)

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“…Abbreviations: CD, cytochalasin D; IP~, inositol trisphosphste ; DG, diacylglycerol; CCh, carbachol; ISO, isoproterenol. no effect on protein secretion induced by the activation of cholinergic mffscarinic receptors [6,11,25].…”

Section: Introductionmentioning

confidence: 98%

Microfilaments and cellular signal transduction: Effect of cytochalasin D on the production of cAMP, inositol phosphates, and on calcium movements in rat parotid glands

Huleux

Dreux

Imhoff

et al. 1989

Biology of the Cell

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In rat parotid glands, the involvement of the microfilament system in the cellular signal transmission mechanism was tested by measuring the effect of cytochalasin D (which disturbs the microfilament system) on the production of intracellular second messengers. Cytochalasin D (CD) did not affect unstimulated calcium movements (measured by the 45Ca efflux technique) or inositol phosphate production or cAMP accumulation. Neither did it modify the generation of intracellular second messengers induced by activation of the cholinergic muscarinic receptor (calcium and inositol phosphates). CD dit not affect the cAMP accumulation induced by the activation of the beta-adrenergic receptor whereas it strongly inhibited the calcium movements induced by activation of the same receptor. These data suggest that, in rat parotid glands, calcium movements, induced by beta-adrenergic receptor stimulation need an intact microfilament system to occur, whereas the muscarinic pathway (via IP3) does not.

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Section: Introductionmentioning

confidence: 98%

Microfilaments and cellular signal transduction: Effect of cytochalasin D on the production of cAMP, inositol phosphates, and on calcium movements in rat parotid glands

Huleux

Dreux

Imhoff

et al. 1989

Biology of the Cell

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“…As a matter of fact, carbachol, which is able to induce calcium entry in the cell even in the presence of cytochalasin D [19] partially abolished the inhibitory effect of cytochalasin D. Furthermore, the fact that in the absence of calcium in the external medium carbachol is not able to abolish the inhibitory effect of cytochaiasin D, indicates that the intracellular calcium mobilized by carbachol (via IP3) is not sufficient to restore ~-adrenergic secretion. We also have shown that, when isoproterenol was present as the same time as cytochalasin D, protein granules were "discharged in the vacuoles formed" by cytochalasin D [8]. In parotid glands cytochaiasin D would inhibit p-adrenergic induced secretion only because it inhibits p-adrenergic calcium movements [19].…”

Section: Discussionmentioning

confidence: 86%

“…These results lead us to suggest that the inhibitory effect of cytochalasin D could be abolished by a calcium entry in the cell. Actually, we had already shown that cytochalasin D induced the formation of empty vacuoles [8]. These results lead us to think that there would be no correlation between actin reorganization and the re-establishment of protein secretion.…”

Section: Discussionmentioning

confidence: 97%

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Process of re‐establishment of β‐adrenergic induced protein secretion after cytochalasin D inhibition in rat parotid gland. Effect of cholinergic agonist, phorbol ester and calcium

Huleux¹,

Dreux²,

Lemullois³

et al. 1991

Biology of the Cell

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In rat parotid gland, 3H-protein secretion is stimulated by beta-adrenergic receptor activation (via cAMP) and also by cholinergic receptor activation (via IP3, calcium and diacylglycerol). The disorganization of microfilament system by cytochalasin D induced an inhibition of beta-adrenergic induced 3H-protein secretion whereas it did not modify the cholinergic muscarinic one. Cytochalasin D induced the formation of vacuoles in the parotid cell. In this work we show that the activation of muscarinic receptors (with carbachol) partially abolished the inhibitory effect of cytochalasin D on beta-adrenergic induced secretion. Since carbachol induced both intracellular calcium increase and protein kinase C activation, we decided to test separately the effect of calcium (using the calcium ionophore A23187) and protein kinase C activation (using phorbol ester) on the inhibitory effect of cytochalasin D on beta-adrenergic induced secretion. A23187, in the presence of calcium in the external medium was able to partially abolish cytochalasin D effect (ie re-establishing protein secretion) whereas activation of protein kinase C by phorbol 12-13 di-butyrate had no effect. These results suggest that protein kinase C is not involved in re-establishing a 'normal' secretion phenomenon whereas calcium does interfere. Furthermore, our fluorescence study shows that, when cytochalasin D is present in the incubation medium, the actin network is disturbed even in the presence of carbachol. This indicates that a calcium entry in the cell is not sufficient to restore a 'normal' actin network.(ABSTRACT TRUNCATED AT 250 WORDS)

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“…In earlier papers (5,8,15) in which colchicine (which forms the irreversible complex with tubulin) was used it was shown that (i) destruction of microtubules slowed down the passage of proteins in the ribosomal endoplasmic reticulum-Golgi area; and (ii) secretion granules formed in the presence of colchicine were not discharged. We could not, however, establish the stage at which the secretory granules underwent the modification that made them incapable of exocytosis.…”

Section: Discussionmentioning

confidence: 99%

Colchicine analogues that bind reversibly to tubulin define microtubular requirements for newly synthesized protein secretion in rat lacrimal gland.

Herman

Busson

Gorbunoff

et al. 1989

Proc. Natl. Acad. Sci. U.S.A.

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The role of microtubules in 3H-labeled protein secretion in rat lacrimal glands was probed by the use of colchicine and two of its analogues that reversibly bind to tubulin. These analogues were 2-methoxy-5-(2,3,4-trimethoxyphenyl)-2,4,6-cycloheptatriene-1-one and 2,3,4,4'-tetramethoxy-1,1'-biphenyl, the latter having been synthesized for these studies. Immunofluorescence revealed that untreated exocrine acinar cells contained an intact microtubule network, which was totally abolished by drug addition. Subsequent drug removal restored the network for the two reversibly binding drugs-more rapidly so for the biphenyl, but this was not the case with colchicine. The protein-secretory process was examined by adding the three drugs at various stages-prepulse incubation, pulse, maturation, apical storage of granules, and discharge under cholinergic stimulation. Comparison with the kinetics of microtubular network restoration, which differed for the two reversibly binding drugs, led to the conclusion that the microtubular system is critical to the maturation phase of secretion.The practical irreversibility of colchicine binding to tubulin (1) limits considerably the use of this drug as a probe of cellular mechanisms because its effects are essentially allor-none in character. Recently, several analogues of colchicine have been synthesized (refs. 2 and 4; M.J.G., unpublished results) that bind reversibly to pure tubulin in the colchicine-binding site and inhibit microtubule assembly reversibly at substoichiometric levels (3, §). It appeared, therefore, that these colchicine analogues should be much more sensitive probes of the linkage between protein secretoryprocess stages and microtubular integrity than the parent drug. In previous studies (5), we proposed that perturbation of the microtubular network by colchicine induces a slowdown of secretory-protein transport from ribosomal endoplasmic reticulum to Golgi region and inhibition of the discharge of secretory granules formed under these conditions. Therefore, microtubules could interfere with the steps offormation, maturation, and apical storage of new secretory granules. From our earlier hypothesis, addition of the reversibly bound colchicine analogues would be predicted to disrupt the cellular microtubule network and consequently arrest the secretory process. Removal of these drugs should reverse such effects if inhibition is caused solely by the absence of microtubules.To test this hypothesis parallel studies were undertaken on the effects of such colchicine analogues on (i) integrity of the microtubule network in acini and (ii) exocrine secretion of protein in lobules from rat exorbital lacrimal glands. The results obtained with two drugs that are progressively simplified analogues of colchicine are reported in this paper. The drugs are 2-methoxy-5-(2,3,4-trimethoxyphenyl)-2,4,6-cycloheptatriene-1-one (MTC) (2) (structure II) and 2,3,4,4'-tetramethoxy-1,1'-biphenyl (TMB) (M.J.G., unpublished results) (structure III). MTC is the colchicine molecule from whi...

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Cytochalasin D suppresses beta‐adrenergic induced protein discharge without inhibiting membrane fusion

Cited by 6 publications

References 0 publications

Microfilaments and cellular signal transduction: Effect of cytochalasin D on the production of cAMP, inositol phosphates, and on calcium movements in rat parotid glands

Microfilaments and cellular signal transduction: Effect of cytochalasin D on the production of cAMP, inositol phosphates, and on calcium movements in rat parotid glands

Process of re‐establishment of β‐adrenergic induced protein secretion after cytochalasin D inhibition in rat parotid gland. Effect of cholinergic agonist, phorbol ester and calcium

Colchicine analogues that bind reversibly to tubulin define microtubular requirements for newly synthesized protein secretion in rat lacrimal gland.

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