The cytoplasmic matrix of the adrenal chromaffin cells of rats under normal and stressed conditions.
ABSTRACT In embedment-free electron microscopy with polyethylene glycol embedding and subsequent deembedding, the conventional cytoplasm of the chromaffin cells was revealed to consist of a three-dimensional lattice of microtrabeculae and gives the impression that the chromaffin granules are held in place by the lattice. After the restraint stress, a substantial number of chromaffin cells were almost free of granules, and the microtrabecular lattice was much more compact than that in cytoplasmic regions occupied with remaining granules or increased mitochondria. In immunocytochemistry, actin immunofluorescence was confined to the subplasmalemmal regions, while tubulin and tropomyosin immunofluorescence appeared throughout the entire cytoplasm of normal chromaffin cells. After the stress, the immunofluorescence for actin and tubulin increased in intensity, while that for tropomyosin decreased. Immunogold labelings for actin and tubulin were found mainly on the thinner subplasmalemmal microtrabeculae and the thicker perikaryal ones, respectively, while some were deposited in the form of small aggregates on portions of microtrabeculae. No specific association between the gold labelings for actin or tubulin and the chromaffin granules was found, even in the subplasmalemmal regions. A hypothetical interpretation was proposed in which a more compact lattice of the microtrabeculae in spatial association with a looser lattice represents a gelated state of the cytoplasm. The significance of the gel-sol transition of the cytoplasmic matrix in relation to the secretory mechanism was discussed.
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ABSTRACT: To compare the effects of cytochalasins on the cellular level with those on the molecular level, 24 cytochalasins, 20 natural compounds and 4 derivatives, were used. The following effects were tested for each of 24 cytochalasins; (a) four high dose (2-20 muM) effects on the cellular level: rounding up of fibroblastic cells, contraction of actin cables, formation of hairy filaments containing actin, and inhibition of lymphocyte capping; (b) a low dose (0.2-2 muM) effect: inhibition of membrane ruffling; and (c) two in vitro effects: an inhibition of actin filament elongation (the high affinity effect [low dose effect] in vitro) and an effect on viscosity of actin filaments(the low affinity effect [high dose effect] in vitro). These results indicated that there are almost the same hierarchic orders of relative effectiveness of different cytochalasins between low and high dose effects and between cellular and molecular effects. From the data obtained with the 24 cytochalasins, we have calculated correlation coefficients of 0.87 and 0.79 between an effect in vivo, inhibition of capping, and an effect in vitro, inhibition of actin filament elongation, as well as between inhibition of capping and another effect in vitro, effect on viscosity of actin filaments, respectively. Furthermore, a correlation coefficient between the high affinity effect and the low affinity effect determined in vitro was calculated to be 0.90 from the data obtained in this study. The strong positive correlation among low and high dose effects in vivo and those in vitro suggests that most of the effects caused by a cytochalasin, irrespective of doses or affected phenomena, might be attributed to the interaction between the drug and the common target protein, actin. In the course of the immunofluorescence microscope study on cytochalasin-treated cells using actin antibody, we have found that aspochalasin D, a 10-isopropylcytochalasin, strongly induced the formation of rodlets containing actin in the cytoplasm of the treated fibroblasts. In contrast, the other cytochalasins, including cytochalasin B, cytochalasin C, cytochalasin D, and cytochalasin H, were found to induce the formation of nuclear rodlets. Both cytoplasmic and nuclear rodlets found in the cytochalasin-treated cells were similar in ultrastructures to those induced by 5 to 10 percent (vol/vol) dimethyl sulfoxide in the same type of cells.The Journal of Cell Biology 02/1982; 92(1):69-78. · 10.82 Impact Factor
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ABSTRACT: Spectrin is an ubiquitous protein composed of heterodimers with alpha and beta subunits. It was first described in erythrocyte cell membranes (see ref. 1 for review) and subsequently in brain, intestinal brush borders, kidney, liver and adrenals. Brain spectrin (fodrin) alpha-subunit, responsible for actin binding, has a relative molecular mass (Mr) of 240,000, whereas the beta-subunit, involved in membrane attachment, has an Mr of 235,000 (refs 1, 3, 9-13). The membrane of secretory granules from adrenal chromaffin cells membrane of secretory granules from adrenal chromaffin cells increases the viscosity of F-actin solution, and spectrin-like protein is associated with storage granule and plasma membranes. Here, we report the localization of fodrin in secretory cells using monospecific antibodies against the alpha-subunit of fodrin using indirect immunofluorescence. We find that the alpha-subunit forms an intensely stained continuous ring in the subplasmalemmal region of resting chromaffin cells. On stimulation of the cell with nicotine, high potassium or ionophores in the presence of calcium, fodrin forms patches. This aggregation is inhibited by trifluoperazine, hence the entrance of calcium into cells following cell depolarization seems to be the calmodulin-dependent stimulus initiating patch formation.Nature 01/1985; 315(6020):589-92. · 38.60 Impact Factor
- Annals of the New York Academy of Sciences 12/2006; 493(1):435 - 447. · 4.38 Impact Factor