Insulin and epidermal growth factor. Human fibroblast receptors related to deoxyribonucleic acid synthesis and amino acid uptake.

Hollenberg,; Cuatrecasas, Pedro

doi:10.1016/s0021-9258(19)41475-0

Cited by 355 publications

(17 citation statements)

References 59 publications

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“…However, it is unclear how these changes might relate to the mitogenic effect of the hormone . Similarly, rapid biochemical alterations have been described in cells in response to EGF, including an increase in a aminoisobutyric acid uptake in human fibroblasts (22), anincrease in 2-deoxyglucose uptake and glycolysis in 3T3 cells (3,15), and an increase in 'Rb+ influx (measure of K+ transport) in 3T3 cells (35). Again, however, we do not know how these early biochemical responses to EGF relate to DNA replication and cell division which occur many hours later .…”

Section: Discussionmentioning

confidence: 90%

Epidermal growth factor induces rapid centrosomal separation in HeLa and 3T3 cells.

Sherline¹,

Mascardo²

1982

The Journal of Cell Biology

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Using indirect immunofluorescence, we have found that epidermal growth factor (EGF), at 100 ng/ml, induces centrosomal separation within 20 min in HeLa and 3T3 cells. The effect was evident both in unsynchronized cultures and in HeLa cells blocked in early S phase by hydroxyurea. EGF also induced centrosomal separation in quiescent 3T3 cells blocked in Go/G 1 by serum deprivation, indicating that DNA replication is not necessary for this effect . The mechanism of this rapid centrosomal separation and its role in the mitogenic effects of EGF remains to be determined .Epidermal growth factor (EGF) (6,045 mol wt) is probably the most extensively studied mammalian mitogenic peptide (6) . EGF was originally isolated from mouse submaxillary glands but it is clearly made by other tissues because submaxillary gland excision does not lower plasma levels of EGF (4) . In man, EGF is found in plasma, saliva, and urine but its precise physiological role is unknown (13, 37). Since a wide variety of cells of epithelial (19,33,39), fibroblastic (1, 2, 5, 21), and neuronal (45) origin are stimulated to proliferate in vitro in response to EGF, it seems that this peptide is likely to be an important regulator of cell division in vivo.As with all other peptide hormones studied thus far, the initial event in the action of EGF is binding to specific highaffinity receptors on the external surface of the plasma membrane (1, 5, 6, 21); 15-22 h later DNA synthesis ensues with subsequent cell replication (1,5,6,22) . Although it is clear that major changes in the organization and function ofthe cell must occur in response to EGF before cell division, the key events leading from membrane binding to cell division are unknown.Centrosomes are perinuclear structures which consist of a pair of centrioles surrounded by an electron-dense cloud of pericentriolar material (34,40). During interphase many cytoplasmic microtubules originate from this structure (14,17,29). During mitosis a centrosome is located at each spindle pole and serves as an anchoring site for microtubules that make up the spindle apparatus . The centriole cycle, which includes duplication, elongation, and polar migration is, in general, believed to be tightly coupled to the cell cycle but the precise relationship between centriole and cell-cycle events remains to be elucidated. For example, while centriole formation can proceed in the absence of DNA synthesis (26, 32) it does require the presence of the nucleus (26). Whether centrosomal behavior influences nuclear events is an open question.Using a specific antiserum to a high molecular weight microtubule-associated protein (MAP,), which stains centrosomes THE JOURNAL OF CELL BIOLOGY " VOLUME 93 MAY 1982 507-511 © The Rockefeller University Press " 0021-9525/82/05/0507/06 $1 .00 in a variety ofcells, we have found that, after addition of EGF, the centrosome splits . The paired daughter centrosomes maintain close contact with the nucleus but migrate away from one another along the nuclear surface . The rapidity ofthe re...

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Section: Discussionmentioning

confidence: 90%

Epidermal growth factor induces rapid centrosomal separation in HeLa and 3T3 cells.

Sherline¹,

Mascardo²

1982

The Journal of Cell Biology

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“…Alteration of the cell surface by trypsinisation [21] was not felt to be a factor in these studies since the cells had been grown to confluence after the previous trypsinisation and were exposed to insulin while in an undisturbed monolayer. It has also been suggested that the fibroblast may not be a primary target tissue for insulin and that its receptor is intended for some other closely related insulin-like polypeptide [2].…”

Section: Discussionmentioning

confidence: 99%

“…A number of metabolic effects of insulin have been described in cultures of fibroblasts obtained from human skin. These effects include the enhancement of the uptake of glucose [1] and of alphaaminoisobutyrate [2], an increase in the incorporation of labelled carbon from glucose into carbon dioxide [3], and the restoration of glycogen levels in glycogen-depleted cells [4]. The incorporation of uridine into RNA and of leucine into protein are also stimulated by the exposure of cultured fibroblasts to insulin [1].…”

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confidence: 99%

Insulin stimulation of glycogen synthase activity in cultured human fibroblasts from diabetic and control subjects

1980

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The influence of insulin on the enzyme glycogen synthase was studied in cultured skin fibroblasts from 5 non-diabetic subjects and 3 diabetic patients. Confluent fibroblast cultures were maintained in serum-free medium for 24 hours before exposure to graduated concentrations of insulin (0.1 to 2000 mU/ml) for one hour. The independent form of glycogen synthase activity (determined in the absence of glucose-6-phosphate) was increased in some experiments by exposure of cells to 0.1mU of insulin/ml; 1.0mU/ml produced a consistent increase; but at least 1000 mU/ml was required for maximum effect. In the non-diabetic subjects, the mean baseline total activity of glycogen synthase was 0.182 ~mol/mg protein/h with 16.8% in the independent form; with 1000 m U of insulin/ml the total activity was 0.167 gmol/mg/h with 45.4% in the independent form. In the diabetic fibroblasts, the mean percentage of glycogen synthase in the independent form was lower than in the control cells at baseline and with all concentrations of insulin utilised; at baseline, total activity was 0.117 ,umol/mg protein/h with 11.7% in the independent form while with 1000mU of insulin/ml the values were 0.146 ~tmol/mg protein/h and 30.7% respectively. Demonstration of an effect of insulin on glycogen synthase in human fibroblasts provides another phenomenon for comparison in cells from control subjects and diabetic patients.

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“…The uptake of 2-deoxyglucose was measured by a modification of the procedure described by Hollenberg and Cuatrecasas (27) for the study of 3-O-methyl-D-glucose transport. PC12 cells cultured on 35-mm tissue culture dishes were rinsed free of medium with warm phosphate-buffered saline (PBS) containing 0.1% bovine serum albumin (BSA).…”

Section: Methodsmentioning

confidence: 99%

Nerve growth factor-induced alteration in the response of PC12 pheochromocytoma cells to epidermal growth factor

Huff¹,

End²,

Guroff³

1981

The Journal of Cell Biology

259

172

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PC12 cells, which differentiate morphologically and biochemically into sympathetic neuronlike cells in response to nerve growth factor, also respond to epidermal growth factor . The response to epidermal growth factor is similar in certain respects to the response to nerve growth factor . Both peptides produce rapid increases in cellular adhesion and 2-deoxyglucose uptake and both induce ornithine decarboxylase . But nerve growth factor causes a decreased cell proliferation and a marked hypertrophy of the cells. In contrast, epidermal growth factor enhances cell proliferation and does not cause hypertrophy . Nerve growth factor induces the formation of neurites ; epidermal growth factor does not.When both factors are presented simultaneously, the cells form neurites . Furthermore, the biological response to epidermal growth factor, as exemplified by the induction of ornithine decarboxylase, is attenuated by prior treatment of the cells with nerve growth factor . PC12 cells have epidermal growth factor receptors. The binding of epidermal growth factor to these receptors is rapid and specific, and exhibits an equilibrium constant of 1 .9 x 10 -9 M. Approximately 80,000 receptors are present per cell, and this number is independent of cell density.Treatment of the cells with nerve growth factor reduces the amount of epidermal growth factor binding by at least 809'0. The decrease in receptor binding begins after -12-18 h of nerve growth factor treatment and is complete within 3 d . Scatchard plots indicate that the number of binding sites decreases, not the affinity of the binding sites for epidermal growth factor .The rat pheochromocytoma clone PC 12 has been used extensively as a model of neuronal differentiation . This clone, developed from an adrenal tumor by Greene and his colleagues (12, 18-20, 22, 45), is of adrenergic neural crest derivation . The cells grow readily under standard conditions and exhibit many of the properties of adrenal medullary chromaffin cells . In the presence of nerve growth factor in nanogram or even subnanogram amounts, the cells differentiate into a phenotype resembling sympathetic neurons . The changes occur within a few days and include process formation, electrical excitability, amine uptake and storage, and a cessation of cell division . The cells contain increased levels of certain transmitter-synthesizing enzymes (13,20,41) and will make synapses with appropriate target tissues (41). The differentiation appears to be reversible, but except for this, it resembles, as far as has been explored, the changes occurring in normal cells differentiating into sympathetic neurons .

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Insulin and epidermal growth factor. Human fibroblast receptors related to deoxyribonucleic acid synthesis and amino acid uptake.

Cited by 355 publications

References 59 publications

Epidermal growth factor induces rapid centrosomal separation in HeLa and 3T3 cells.

Epidermal growth factor induces rapid centrosomal separation in HeLa and 3T3 cells.

Insulin stimulation of glycogen synthase activity in cultured human fibroblasts from diabetic and control subjects

Nerve growth factor-induced alteration in the response of PC12 pheochromocytoma cells to epidermal growth factor

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