Maintenance of pancreatic endocrine cells of the neonatal rat: VII. The effect of 3-amino-3-deoxyglucose on insulin secretion from perifused B cells.

Nakao, Kazuwa; Kagawa, Shohei; Wakabayashi, Shigeru; Matsuoka, Akira

doi:10.1507/endocrj1954.33.561

Cited by 5 publications

(6 citation statements)

References 37 publications

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“…In contrast, 10 mM potassium requires glucose to induce insulin secretion from RIN-38 cells. Forskolin increases the cAMP content of neonatal islets, induces an adultlike insulin secretory response to glucose, and shifts the maximal secretory response to higher concentrations of glucose (9). In the present studies forskolin and other cAMP modulators potentiated secretion, but did not alter the glucose sensitivity of RIN-38 cells; similar results were recently reported with fetal islets (34).…”

Section: Potentiation Of Insulin Secretionsupporting

confidence: 90%

“…Whether in the presence of 10 mM potassium or 0.1 mM IBMX, glucose has an unaltered dose-responsive effect on insulin secretion. As with neonatal islets (9), IBMX in the absence of glucose acts as a secretagogue with RIN-38 cells. In contrast, 10 mM potassium requires glucose to induce insulin secretion from RIN-38 cells.…”

Section: Potentiation Of Insulin Secretionmentioning

confidence: 94%

“…The drawback with these cell lines is that they show a sensitivity to glucose as a secretory signal which is, at best, more typical of immature /3-cells. Normal mature rat islets secrete in response to glucose concentrations greater than 5 mM, with maximal secretion at concentrations of 16 mM or higher (6), while newborn rat islets secrete maximally at lower glucose concentrations (6,8,9). Most /3-cell lines do not secrete insulin in response to glucose or respond maximally at low glucose concentrations (10)(11)(12)(13)(14)(15)(16)(17)(18)(19).…”

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confidence: 99%

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Modulation of Glucose-Induced Insulin Secretion from a Rat Clonal β-Cell Line*

1990

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The present studies demonstrate that the beta-cell line RINr1046-38 (RIN-38) retains the capability to secrete insulin in response to glucose. The maximal effect of glucose was a 5- to 9-fold stimulation of insulin secretion from RIN-38 cells. This glucose-induced insulin secretion was maximal at 0.6 mM and was modulated by other secretagogues. Potassium concentrations of 10 mM, adenylate cyclase activators (glucagon-like peptide-1 and forskolin), and a phosphodiesterase inhibitor (isobutylmethylxanthine) potentiated glucose-induced insulin secretion, but had little or no effect on insulin secretion in the absence of glucose. Potassium concentrations of 20 mM or more, glibenclamide, and carbachol (Cch) stimulated insulin secretion 8- to 12-fold in the absence of glucose, while only Cch potentiated the effect of glucose on insulin secretion. Amino acids (alanine, arginine, leucine, and ketoisocaproate) also stimulated insulin secretion. The alpha 2-adrenergic agonist clonidine (1 microM), low extracellular calcium (less than or equal to 0.5 mM), and extended culture of RIN-38 cells at low glucose concentrations (0.33 mM) inhibited the stimulatory effect of glucose on insulin secretion. Insulin secretion was retained in RIN-38 cells for up to 98 passages. However, extended passage was associated with a decline in cellular insulin content (83% decline over 89 passages). In addition, high passage cells lost the ability to secrete insulin in response to glucose, but continued to respond to other secretagogues (K+, alanine, and carbachol). In fact, in the absence of glucose the effect of Cch on insulin secretion was well maintained in high passage cells (8- and 9.9-fold increase in insulin secretion, passages 9 and 70, respectively). Thus, low passage RIN-38 cells secrete insulin in response to glucose and other insulin secretagogues. High passage cells do not respond to glucose, but continue to respond to other secretagogues. Based on these results we propose that high and low passage RIN-38 cells provide a model for examining molecular mechanisms of glucose-induced insulin secretion. In addition, these findings emphasize that passage information is essential for interpretation of secretion studies with RIN cell lines.

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Section: Potentiation Of Insulin Secretionsupporting

confidence: 90%

Section: Potentiation Of Insulin Secretionmentioning

confidence: 94%

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Modulation of Glucose-Induced Insulin Secretion from a Rat Clonal β-Cell Line*

1990

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“…Perifusion procedure Perifusion of the monolayer cell culture was per¬ formed according to a modification of the method of Kawazu, Kanazawa, Ikeuchi et al (1977), as described previously (Nakao, Kagawa, Wakabayashi & Matsuoka, 1986). In brief, the perifusion chamber was made by placing two discs, made from culture dishes with their rims removed with an electric knife, on both sides of a silicon rubber sheet (thickness 3-5 mm) with a round hole (diameter 3 cm) in the centre.…”

Section: Reagentsmentioning

confidence: 99%

Monolayer cultures of pancreatic islet cells from adult rats: effect of 2-deoxyglucose

Aoki

Kagawa²,

Yamamura³

et al. 1988

Journal of Endocrinology

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Techniques for the monolayer culture of pancreatic islet cells from adult rats and the responsiveness of B cells are described. Whole pancreatic tissue was enzymatically dispersed and then cultured for 30 days in tissue culture medium 199 containing 5.5 mmol glucose/l, with or without 1 mmol 2-deoxyglucose/l. In the absence of 2-deoxyglucose, the responsiveness of B cells diminished to almost zero by day 15 and islets degenerated. In contrast, addition of 2-deoxyglucose to the medium resulted in a selective degeneration of fibroblasts, yielding monolayers that consisted mostly of islet cells. In this stationary system in which monolayers of islet cells were maintained in medium with 2-deoxyglucose, insulin secretion from B cells on days 15 and 30 increased in a dose-dependent fashion in response to increasing concentrations of glucose, leucine and 2-ketoisocaproate. Similarly, when exposed to 16.7 mmol glucose/l, perifused B cells showed a biphasic pattern of insulin secretion on day 15. Addition of 10 mumol forskolin/l and 200 nmol 12-O-tetradecanoyl phorbol-13-acetate/l remarkably enhanced this response. Likewise, the response to 10 mmol leucine/l or 10 mmol 2-ketoisocaproate/l was biphasic. These results suggest that these monolayer cultures retain the functional properties of the adult rat pancreas, and may be useful not only as a model for the in-vitro study of B cell function, but also for implantation.

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“…In conclusion, these results suggest that cultivation in medium with 5\m=.\5mmol glucose/l could induce precocious development of an adult pattern of insulin secretion compared with that in 16\m=.\7mmol glucose/l and, in addition, that the medium with 16\m=.\7mmol INTRODUCTION It is widely acknowledged that differences exist between the response of the fetal, neonatal and adult pancreas to insulin secretagogues. In the fetal and neonatal rat, cells respond poorly to glucose, with little or no rise in the second phase of secretion (Burr, Kanazawa, Marliss & Lambert, 1971 ;Nakao, Kagawa, Wakabayashi & Matsuoka, 1986). In addition, Grill, Lake & Freinkel (1981) have reported that there is a more generalized defect in the recognition of nutrient secretagogues.…”

mentioning

confidence: 99%

Effect of glucose concentrations on functional maturation of monolayer-cultured B cells from the rat pancreas

Wakabayashi

Kagawa²,

Yamashita³

et al. 1988

Journal of Endocrinology

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The effect of two concentrations of glucose on the maturation of the response of B cells was studied in pancreatic monolayer cultures of the neonatal rat using a perifusion system. After exposure for the initial 3 days to a medium with 5.5 mmol glucose/l and 10 mumol iodoacetic acid/l (day 3), in order to delete fibroblasts selectively, monolayer cultures were kept for a total of 12 days in medium with either 5.5 or 16.7 mmol glucose/l alone. On day 3, B cells responded in a monophasic fashion, with no significant second phase, to acute challenge with 16.7 mmol glucose/l. At a concentration of 10 mmol/l, leucine and 2-ketoisocaproate both produced only minimal increases in the second phase of secretion above the basal level. In contrast, B cells on day 7 cultured in 5.5 mmol glucose/l showed a biphasic response to glucose, leucine and 2-ketoisocaproate. The magnitude in response to glucose was well preserved at day 15 of culture, whereas the stimulatory effects of leucine and 2-ketoisocaproate decreased to 24-57% of that of B cells on day 7. Moreover, B cells on day 7 cultured in 16.7 mmol glucose/l responded in a biphasic manner to glucose, the response being 65% of that of B cells in 5.5 mmol glucose/l. Additionally, the response to leucine and 2-ketoisocaproate still appeared to be monophasic. At day 15 of culture, however, the response of B cells in 16.7 mmol glucose/l to glucose was 105%, to leucine 245% and to 2-ketoisocaproate 127% of that of B cells in 5.5 mmol glucose/l.(ABSTRACT TRUNCATED AT 250 WORDS)

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Maintenance of pancreatic endocrine cells of the neonatal rat: VII. The effect of 3-amino-3-deoxyglucose on insulin secretion from perifused B cells.

Abstract: Abstract

Cited by 5 publications

References 37 publications

Modulation of Glucose-Induced Insulin Secretion from a Rat Clonal β-Cell Line*

Modulation of Glucose-Induced Insulin Secretion from a Rat Clonal β-Cell Line*

Monolayer cultures of pancreatic islet cells from adult rats: effect of 2-deoxyglucose

Effect of glucose concentrations on functional maturation of monolayer-cultured B cells from the rat pancreas

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