Affinity purified tetanus toxin binds to isolated chromaffin granules and inhibits catecholamine release in digitonin‐permeabilized chromaffin cells

Lazarovici, Philip; Fujita, Kanna; Contreras, Margarita L.; DiOrio, James P.; Lelkes, Peter I.

doi:10.1016/0014-5793(89)80943-3

Cited by 16 publications

(6 citation statements)

References 30 publications

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“…When the toxin's binding and internalization are modified by ganglioside loading (Marxen et al, 1989) or membrane permeabilization (Bittner and Holz, 1988;Ahnert-Hilger et al, 1989;Bittner et al, 1989;Lazarovici et al, 1989), normally nonreceptive cells show toxin inhibition of exocytosis. Direct microinjection of the toxin (Penner et al, 1986;Mochida et al, 1989;Poulain et al, 1990) results in very rapid effects.…”

Section: Discussionmentioning

confidence: 99%

Differential Effects of Tetanus Toxin on Inhibitory and Excitatory Neurotransmitter Release from Mammalian Spinal Cord Cells in Culture

Williamson

Fitzgerald

Neale

1992

Journal of Neurochemistry

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The effect of tetanus toxin on depolarization‐evoked and spontaneous synaptic release of inhibitory and excitatory neurotransmitters was examined in murine spinal cord cell cultures. Toxin action on the release of radiolabeled glycine and glutamate was followed over time intervals corresponding to the early phase of convulsant activity through the later phase of electrical quiescence. Tetanus toxin inhibited potassium‐evoked release of [3H]glycine and [3H]glutamate in a time‐ and dose‐dependent manner. Ninety minutes after the application of toxin (6 × 10−10M), the stimulated release of [3H]glycine was blocked completely, whereas stimulated release of [3H]glutamate was not blocked completely until 150–210 min after toxin application. Fragment C, the binding portion of the tetanus toxin molecule, had no effect on stimulated release of either transmitter. The spontaneous synaptic release of [3H]glycine was blocked totally within 90 min of toxin exposure. In contrast, the spontaneous release of [3H]glutamate, in toxin‐exposed cultures, was elevated to nearly twice that of control cultures at this time. Thus, toxin‐induced convulsant activity is characterized by a reduction in the spontaneous synaptic release of inhibitory neurotransmitter with a concomitant increase in the release of excitatory neurotransmitter, as well as the more rapid onset of blockade of depolarization‐evoked release of inhibitory versus excitatory neurotransmitter.

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Section: Discussionmentioning

confidence: 99%

Differential Effects of Tetanus Toxin on Inhibitory and Excitatory Neurotransmitter Release from Mammalian Spinal Cord Cells in Culture

Williamson

Fitzgerald

Neale

1992

Journal of Neurochemistry

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“…The ultrastructure of 3-D cultures maintained in vitro for 4 weeks in Matrigel gel and electrospun fibrous scaffolds, respectively, were examined as previously described. 17 Briefly, the samples were washed with cold 0.2 M Na cacodylate buffer (pH 7.4) and fixed overnight with 2.5% gluteraldehyde in cacodylate buffer. Following an additional wash in buffer, the samples were post-fixed with 2% OsO 4 in cacodylate buffer (1 h, 4°C), dehydrated in graded concentrations of cold ethyl alcohol, and embedded in epoxy resin.…”

Section: Transmission Electron Microscopymentioning

confidence: 99%

Engineering Three-Dimensional Pulmonary Tissue Constructs

et al. 2006

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In this paper, we report on engineering 3-D pulmonary tissue constructs in vitro. Primary isolates of murine embryonic day 18 fetal pulmonary cells (FPC) were comprised of a mixed population of epithelial, mesenchymal, and endothelial cells as assessed by immunohistochemistry and RT-PCR of 2-D cultures. The alveolar type II (AE2) cell phenotype in 2-D and 3-D cultures was confirmed by detection of SpC gene expression and presence of the gene product prosurfactant protein C. Three-dimensional constructs of FPC were generated utilizing Matrigel hydrogel and synthetic polymer scaffolds of poly-lactic-co-glycolic acid (PLGA) and poly-L-lactic-acid (PLLA) fabricated into porous foams and nanofibrous matrices, respectively. Three-dimensional Matrigel constructs contained alveolar forming units (AFU) comprised of cells displaying AE2 cellular ultrastructure while expressing the SpC gene and gene product. The addition of tissue-specific growth factors induced formation of branching, sacculated epithelial structures reminiscent of the distal lung architecture. Importantly, 3-D culture was necessary for inducing expression of the morphogenesis-associated distal epithelial gene fibroblast growth factor receptor 2 (FGFr2). PLGA foams and PLLA nanofiber scaffolds facilitated ingrowth of FPC, as evidenced by histology. However, these matrices did not support the survival of distal lung epithelial cells, despite the presence of tissue-specific growth factors. Our results may provide the first step on the long road toward engineering distal pulmonary tissue for augmenting and/or replacing dysfunctional native lung in diseases, such as neonatal pulmonary hypoplasia.

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“…Tetanus and botulinum neurotoxins can inhibit exocytosis in permeabilized chromaffin or PC12 cells (Bittner et al, 1989a,b;Ahnert-Hilger et al, 1989a,b;Mochida et al, 1989;Lazarovici et al, 1989;Ahnert-Hilger et al, 1992) or when injected via patch pipette (Penner et al, 1986). A major finding of these studies was that the ability of clostridial neurotoxins to inhibit secretion is confined to the light-chain region of these proteins.…”

Section: Introductionmentioning

confidence: 99%

Protein kinase C and clostridial neurotoxins affect discrete and related steps in the secretory pathway

Bittner

Holz

1993

Cell Mol Neurobiol

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1. The effects on catecholamine secretion of activation of protein kinase C and clostridial neurotoxins were examined in digitonin-permeabilized bovine adrenal chromaffin cells. 2. The enhancement by phorbol esters increased only the initial rate of secretion; later rates were unaffected. This enhancement was present over a wide range of Ca2+ concentrations and was elicited at 18 as well as at 27 degrees C. 3. Tetanus toxin inhibited both ATP-dependent and ATP-independent secretion, indicating that the tetanus toxin target is important during the final steps in the pathway. 4. Prior activation of protein kinase C by the phorbol ester 12-O-tetradecanoyl phorbol acetate rendered the primed state more sensitive to inhibition by tetanus toxin. The data indicate that a phosphorylated protein kinase C substrate is either identical to or closely associated with the tetanus toxin target protein at the final steps in the pathway. 5. The interaction between the effect of protein kinase activation and that of tetanus toxin suggests that protein kinase C activation does not stimulate a separate pathway of secretion but, rather, modulates the activity of the ongoing pathway. 6. The enhancement of secretion by protein kinase C is caused, at least in part, by a qualitative change in the characteristics of the primed state. This is indicated by the increased sensitivity of primed secretion to inhibition by tetanus toxin and a threefold increase in sensitivity of primed secretion to Ca2+. 7. Because activation of protein kinase C does not increase the later rates of secretion that are limited by ATP-dependent priming reactions, it is unlikely that enhancement of the maximal rate of secretion by TPA is due to an increased amount of the primed state. Instead, protein kinase C activation may increase the efficacy with which Ca2+ stimulates secretion at all Ca2+ concentrations.

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Affinity purified tetanus toxin binds to isolated chromaffin granules and inhibits catecholamine release in digitonin‐permeabilized chromaffin cells

Cited by 16 publications

References 30 publications

Differential Effects of Tetanus Toxin on Inhibitory and Excitatory Neurotransmitter Release from Mammalian Spinal Cord Cells in Culture

Differential Effects of Tetanus Toxin on Inhibitory and Excitatory Neurotransmitter Release from Mammalian Spinal Cord Cells in Culture

Engineering Three-Dimensional Pulmonary Tissue Constructs

Protein kinase C and clostridial neurotoxins affect discrete and related steps in the secretory pathway

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