Regulation of Cell Motility by Cyclic AMP

Johnson, George S.; Morgan, Willie D.; Pastan, Ira

doi:10.1038/235054a0

Cited by 164 publications

(34 citation statements)

References 19 publications

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“…The demonstration that phosphate addition decreases the steady-state level of cAMP provides a possible explanation for these effects. Thus, the stimulation of cell migration by phosphate (10) is consistent with a role of cAMP in control of movement (16) and with the effect of phosphate addition on cAMP levels reported here.…”

Section: Discussionsupporting

confidence: 72%

Kinetics of Early Changes in Phosphate and Uridine Transport and Cyclic AMP Levels Stimulated by Serum in Density-Inhibited 3T3 Cells

Asúa

Rozengurt

Dulbecco

1974

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

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A kinetic study of the early changes in uridine-and phosphate-transport rates and 3':5'-cyclic AMP levels induced by the addition of serum to quiescent 3T3 cells revealed that the increase in phosphate uptake and the decrease in the intracellular concentration of 3': 5'-cyclic AMP immediately follow the addition of serum. In contrast, the increase in uridine transport occurred after a lag of several minutes and was clearly preceded by the other changes. The activation of phosphate transport involved an increase in V.., while a more complex pattern was observed for uridine transport. A possible role of phosphate in connection with the other early events was also studied. The addition of phosphate to quiescent cells produced a time-dependent decrease in 3': 5'-cyclic AMP levels and this ion was required for full activation of uridine transport by serum. The results show that the increase in phosphate transport is a primary event in the reinitiation of growth and suggest that the increase in phosphate transport may be connected with subsequent metabolic steps.Addition of serum to density-inhibited 3T3 cells rapidly stimulates several changes in membrane-dependent properties (1) and subsequently promotes reinitiation of DNA synthesis and cell division (2). Within minutes of serum addition, phosphate-and uridine-transport systems are increased several fold (3), and 3':5'-cyclic AMP (cAMP), which is strongly implicated in the regulation of growth and morphology of cultured fibroblasts, decreases (4-6). cAMP may play an important role in the changes of several transport systems (7), and it strongly affects the early activation of uridine transport by serum or insulin (8). However, the stimulation of phosphate transport by serum is affected only slightly by changes of cAMP concentration (8), suggesting that this activation is achieved by a cAMP-independent mechanism. Therefore, phosphate transport is of particular interest because it may be a primary event in the regulation of cell growth. This possibility is also suggested by the observation that the saturation density (9) and the migration of 3T3 cells (10) are influenced by the concentration of this ion in the medium.In this paper, we report on the activation of phosphate transport in density-inhibited 3T3 cells by serum, defining its relationship to other early events such as changes of cAMP levels and of uridine transport activity. MATERIALS AND METHODSBALB/c/3T3 cells (Clone A31) (17) were maintained in reinforced Eagle's medium supplemented with 10% serum, 100 units/ml of penicillin, and 100 /Ag/ml of streptomycin in Nunc petri dishes. Transport was measured at 370 on cells attached to 30-mm dishes. Cultures of confluent cells were usually used 3 days after the last change of medium. In order to measure uptake of labeled substrates into acid-soluble pools, the cells 96 were washed four times with ice-cold isotonic saline and extracted for 30 min at 40 with 5% trichloroacetic acid. Radioactivity was measured in an aliquot counted in Triton-toluene scintill...

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Section: Discussionsupporting

confidence: 72%

Kinetics of Early Changes in Phosphate and Uridine Transport and Cyclic AMP Levels Stimulated by Serum in Density-Inhibited 3T3 Cells

Asúa

Rozengurt

Dulbecco

1974

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

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“…The inverse correlation between adhesiveness of the various cell types to substrate and their apparent responsiveness to CD may also be related to the differential responsiveness of individual cells . It is pertinent that adhesivity of monolayers is greatly increased by the addition of dibutyryl cyclic AMP (44) and their mobility is correspondingly diminished (46) . Prior application of dibutyryl cyclic AMP, under appropriate conditions, markedly diminishes the sensitivity of cell lines such as HeLa to the contractile (60) and zeiotic effects of CD (60,75) .…”

Section: Dif Ferential Sensitivitymentioning

confidence: 99%

Action of Cytochalasin D on Cells of Established Lines

Miranda

Godman

Deitch

et al. 1974

The Journal of Cell Biology

216

119

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HeLa, Vero, L, HEp2, and MDBK cells respond immediately to 0.2–0.5 µg/ml cytochalasin D (CD) with sustained contraction (contracture), loss of microvilli, expression of endoplasmic contents (zeiosis), nuclear protrusion, and extension of cytoplasmic processes. The development of these changes is depicted, and the dose-response patterns in these cell lines are described. MDBK is generally most resistant and HeLa most sensitive to these effects of CD. Cells in G1 are most sensitive to CD; responsiveness decreases progressively during early S and is least in mid S through G2. CD inhibits transport of [14C]deoxyglucose in HeLa by about 45% but has no significant effect on hexose uptake in Vero and MDBK; sugar transport is thus apparently unrelated to any morphologic effect of CD. Although spreading and attachment are impeded, CD does not decrease and may even enhance the adhesiveness of established monolayers. Contraction appears to be a primary early effect of CD, upon which other visible changes follow. It is prevented by some inhibitors of energy metabolism (deoxyglucose and dinitrophenol) and does not occur in glycerinated models without ATP. The possible bases of the contractile response to CD are discussed. Although direct or indirect action of CD on some microfilaments may occur, a generalized structural disruption of contractile filaments by CD is considered unlikely.

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“…Adenosine 3': 5'-monophosphate (cAMP) regulates numerous functions of cultured fibroblastic cells. Among these are growth rate (1)(2)(3)(4)(5), cell shape (2,(6)(7)(8), adhesiveness (9), motility (10), and agglutination by plant lectins such as concanavalin A (Con A) (11)(12)(13)(14). Indeed, when transformed cells are treated with cAMP analogues such as dibutyryl cAMP (Bt2cAMP), their agglutinability is markedly diminished (11)(12)(13)(14).…”

mentioning

confidence: 99%

Cyclic AMP modulates microvillus formation and agglutinability in transformed and normal mouse fibroblasts.

Pastan¹

1975

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

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We have utilized dark field microscopy to observe the surface microstructure of living cultured cells. Using this method, we have found that dibutyryl cAMP treatment causes regression of the numerous, long cell surface microvilli present on L929 cells. Thirty minutes after removal of dibutyryl cAMP, microvilli reappear. An inhibitor of phosphodiesterase (methylisobutylxanthine) and a stimulator of adenylate cyclase (prostaglandin El), both of which raise cAMP levels, cause regression of microvilli in 15 min. Untransformed 3T3 cells show very few microvilli when viewed still attached to their substratum or after removal with EDTA. Treatment of these cells with trypsin causes the formation of numerous microvilli on their surface.. When clumps of cells agglutinated by concanavalin A are examined by thin section electron microscopy, the cells are seen to be held together by a "forest" of interdigitating microvilli and only rarely is there apposition of the areas of membrane between microvilli. At the same time the distribution of surfacebound concanavalin A was examined using immunofluorescent light microscopy, and concanavalin A was found to be uniformly distributed over the cell surface. We propose that agglutinability of mouse and rat fibroblasts is regulated through the modulation of cell surface microvilli by cAMP, and that transformed cells are highly agglutinable because their low cAMP levels result in the formation of numerous surface microvilli. Adenosine 3': 5'-monophosphate (cAMP) regulates numerous functions of cultured fibroblastic cells. Among these are growth rate (1-5), cell shape (2, 6-8), adhesiveness (9), motility (10), and agglutination by plant lectins such as concanavalin A (Con A) (11)(12)(13)(14). Indeed, when transformed cells are treated with cAMP analogues such as dibutyryl cAMP (Bt2cAMP), their agglutinability is markedly diminished (11)(12)(13)(14) MATERIALS AND METHODS Examination of Cells by Light Microscopic Methods. L929or 3T3-4 cells were grown on 22 X 40 mm sterile glass coverslips in 20 cm2 plastic dishes in Dulbecco-Vogt's modified Eagle's medium supplemented with 10% calf serum and penicillin-streptomycin (50 units/ml each) at 370 in 95% air-5% CO2. Preparation and addition of Bt2cAMP, 1-methyl-3-isobutylxanthine, and prostaglandin El were performed as previously described (8). Cells on coverslips were viewed either under phase contrast microscopy or dark field, visible light microscopy on a microscope stage in a plastic enclosure containing 95% air, 5% CO2. The plastic enclosure was kept in a constant temperature room at 37°. Coverslips were removed from their dishes and overlaid with a second sterile coverslip. This sandwich with cells in between was placed on the stage of a Zeiss RA or inverted phase contrast microscope. For dark field observation, a dark field oil condenser (Zeiss ultracondenser) and a 40 X oil immersion lens with an integral iris diaphragm were used. The light source for this observation was a dc-powered mercury vapor light source (HBO 200W/4) u...

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Regulation of Cell Motility by Cyclic AMP

Cited by 164 publications

References 19 publications

Kinetics of Early Changes in Phosphate and Uridine Transport and Cyclic AMP Levels Stimulated by Serum in Density-Inhibited 3T3 Cells

Kinetics of Early Changes in Phosphate and Uridine Transport and Cyclic AMP Levels Stimulated by Serum in Density-Inhibited 3T3 Cells

Action of Cytochalasin D on Cells of Established Lines

Cyclic AMP modulates microvillus formation and agglutinability in transformed and normal mouse fibroblasts.

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