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Endocrinology, Vol 130, 167-178, Copyright © 1992 by Endocrine Society

ARTICLES

Establishment of 2-mercaptoethanol-dependent differentiated insulin- secreting cell lines

M Asfari, D Janjic, P Meda, G Li, PA Halban and CB Wollheim
Department of Medicine, University of Geneva, Switzerland.

New insulin-secreting cell lines (INS-1 and INS-2) were established from cells isolated from an x-ray-induced rat transplantable insulinoma. The continuous growth of these cells was found to be dependent on the reducing agent 2-mercaptoethanol. Removal of this thiol compound caused a 15-fold drop in total cellular glutathione levels. These cells proliferated slowly (population doubling time about 100 h) and, in general, showed morphological characteristics typical of native beta-cells. Most cells stained positive for insulin and did not react with antibodies against the other islet hormones. The content of immunoreactive insulin was about 8 micrograms/10(6) cells, corresponding to 20% of the native beta-cell content. These cells synthesized both proinsulin I and II and displayed conversion rates of the two precursor hormones similar to those observed in rat islets. However, glucose failed to stimulate the rate of proinsulin biosynthesis. In static incubations, glucose stimulated insulin secretion from floating cell clusters or from attached cells. Under perifusion conditions, 10 mM but not 1 mM glucose enhanced secretion 2.2-fold. In the presence of forskolin and 3-isobutyl-1-methylxanthine, increase of glucose concentration from 2.8-20 mM caused a 4-fold enhancement of the rate of secretion. Glucose also depolarized INS-1 cells and raised the concentration of cytosolic Ca2+. This suggests that glucose is still capable of eliciting part of the ionic events at the plasma membrane, which leads to insulin secretion. The structural and functional characteristics of INS-1 cells remained unchanged over a period of 2 yr (about 80 passages). Although INS-2 cells have not been fully characterized, their insulin content was similar to that of INS-1 cells and they also remain partially sensitive to glucose as a secretagogue. INS-1 cells retain beta-cell surface antigens, as revealed by reactivity with the antigangloside monoclonal antibodies R2D6 and A2B5. These findings indicate that INS-1 cells have remained stable and retain a high degree of differentiation which should make them a suitable model for studying various aspects of beta-cell function.


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S. Cheviet, P. Bezzi, R. Ivarsson, E. Renstrom, D. Viertl, S. Kasas, S. Catsicas, and R. Regazzi
Tomosyn-1 is involved in a post-docking event required for pancreatic {beta}-cell exocytosis
J. Cell Sci., July 15, 2006; 119(14): 2912 - 2920.
[Abstract] [Full Text] [PDF]

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 Am. J. Physiol. Endocrinol. Metab.Home page
N. Sekine, K. Takano, N. Kimata-Hayashi, T. Kadowaki, and T. Fujita
Adrenomedullin inhibits insulin exocytosis via pertussis toxin-sensitive G protein-coupled mechanism.
Am J Physiol Endocrinol Metab, July 1, 2006; 291(1): E9 - E14.
[Abstract] [Full Text] [PDF]

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 J. Pharmacol. Exp. Ther.Home page
G. Liu, S. M. P. Jacobo, N. Hilliard, and G. H. Hockerman
Differential Modulation of Cav1.2 and Cav1.3-Mediated Glucose-Stimulated Insulin Secretion by cAMP in INS-1 Cells: Distinct Roles for Exchange Protein Directly Activated by cAMP 2 (Epac2) and Protein Kinase A
J. Pharmacol. Exp. Ther., July 1, 2006; 318(1): 152 - 160.
[Abstract] [Full Text] [PDF]

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 EndocrinologyHome page
L.-X. Li, P. E. MacDonald, D. S. Ahn, G. Y. Oudit, P. H. Backx, and P. L. Brubaker
Role of Phosphatidylinositol 3-Kinase{gamma} in the {beta}-Cell: Interactions with Glucagon-Like Peptide-1
Endocrinology, July 1, 2006; 147(7): 3318 - 3325.
[Abstract] [Full Text] [PDF]

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 EndocrinologyHome page
E. Karaskov, C. Scott, L. Zhang, T. Teodoro, M. Ravazzola, and A. Volchuk
Chronic Palmitate But Not Oleate Exposure Induces Endoplasmic Reticulum Stress, Which May Contribute to INS-1 Pancreatic {beta}-Cell Apoptosis
Endocrinology, July 1, 2006; 147(7): 3398 - 3407.
[Abstract] [Full Text] [PDF]

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 J. Physiol.Home page
G. Kang, O. G. Chepurny, B. Malester, M. J. Rindler, H. Rehmann, J. L. Bos, F. Schwede, W. A. Coetzee, and G. G. Holz
cAMP sensor Epac as a determinant of ATP-sensitive potassium channel activity in human pancreatic {beta} cells and rat INS-1 cells
J. Physiol., June 15, 2006; 573(3): 595 - 609.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
M. K. Lingohr, I. Briaud, L. M. Dickson, J. F. McCuaig, C. Alarcon, B. L. Wicksteed, and C. J. Rhodes
Specific Regulation of IRS-2 Expression by Glucose in Rat Primary Pancreatic Islet beta-Cells
J. Biol. Chem., June 9, 2006; 281(23): 15884 - 15892.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
X. Yang, P. Xu, Y. Xiao, X. Xiong, and T. Xu
Domain Requirement for the Membrane Trafficking and Targeting of Syntaxin 1A
J. Biol. Chem., June 2, 2006; 281(22): 15457 - 15463.
[Abstract] [Full Text] [PDF]

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 J Mol EndocrinolHome page
J. Vikman, X. Ma, G. H Hockerman, P. Rorsman, and L. Eliasson
Antibody inhibition of synaptosomal protein of 25 kDa (SNAP-25) and syntaxin 1 reduces rapid exocytosis in insulin-secreting cells.
J. Mol. Endocrinol., June 1, 2006; 36(3): 503 - 515.
[Abstract] [Full Text] [PDF]

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 DiabetesHome page
V. Cirulli, J. Zalatan, M. McMaster, R. Prinsen, D. R. Salomon, C. Ricordi, B. E. Torbett, P. Meda, and L. Crisa
The Class I HLA Repertoire of Pancreatic Islets Comprises the Nonclassical Class Ib Antigen HLA-G.
Diabetes, May 1, 2006; 55(5): 1214 - 1222.
[Abstract] [Full Text] [PDF]

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 DiabetesHome page
L. I. Rachek, N. P. Thornley, V. I. Grishko, S. P. LeDoux, and G. L. Wilson
Protection of INS-1 Cells From Free Fatty Acid-Induced Apoptosis by Targeting hOGG1 to Mitochondria.
Diabetes, April 1, 2006; 55(4): 1022 - 1028.
[Abstract] [Full Text] [PDF]

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 Proc. Natl. Acad. Sci. USAHome page
P. A. Antinozzi, A. Garcia-Diaz, C. Hu, and J. E. Rothman
Functional mapping of disease susceptibility loci using cell biology.
PNAS, March 7, 2006; 103(10): 3698 - 3703.
[Abstract] [Full Text] [PDF]

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 J. Clin. Endocrinol. Metab.Home page
P. de Lonlay, A. Simon-Carre, M.-J. Ribeiro, N. Boddaert, I. Giurgea, K. Laborde, C. Bellanne-Chantelot, V. Verkarre, M. Polak, J. Rahier, et al.
Congenital Hyperinsulinism: Pancreatic [18F]Fluoro-L-Dihydroxyphenylalanine (DOPA) Positron Emission Tomography and Immunohistochemistry Study of DOPA Decarboxylase and Insulin Secretion
J. Clin. Endocrinol. Metab., March 1, 2006; 91(3): 933 - 940.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
A. L. Garcia, S.-K. Han, W. G. Janssen, Z. Z. Khaing, T. Ito, M. J. Glucksman, D. L. Benson, and S. R. J. Salton
A Prohormone Convertase Cleavage Site within a Predicted {alpha}-Helix Mediates Sorting of the Neuronal and Endocrine Polypeptide VGF into the Regulated Secretory Pathway
J. Biol. Chem., December 16, 2005; 280(50): 41595 - 41608.
[Abstract] [Full Text] [PDF]

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 Mol. Endocrinol.Home page
L. Waselle, R. R. L. Gerona, N. Vitale, T. F. J. Martin, M.-F. Bader, and R. Regazzi
Role of Phosphoinositide Signaling in the Control of Insulin Exocytosis
Mol. Endocrinol., December 1, 2005; 19(12): 3097 - 3106.
[Abstract] [Full Text] [PDF]

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 DiabetesHome page
K. Laubner, T. J. Kieffer, N. T. Lam, X. Niu, F. Jakob, and J. Seufert
Inhibition of Preproinsulin Gene Expression by Leptin Induction of Suppressor of Cytokine Signaling 3 in Pancreatic {beta}-Cells
Diabetes, December 1, 2005; 54(12): 3410 - 3417.
[Abstract] [Full Text] [PDF]

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 J. Cell Sci.Home page
F. Allagnat, D. Martin, D. F. Condorelli, G. Waeber, and J.-A. Haefliger
Glucose represses connexin36 in insulin-secreting cells
J. Cell Sci., November 15, 2005; 118(22): 5335 - 5344.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
H. Iguchi, Y. Ikeda, M. Okamura, T. Tanaka, Y. Urashima, H. Ohguchi, S. Takayasu, N. Kojima, S. Iwasaki, R. Ohashi, et al.
SOX6 Attenuates Glucose-stimulated Insulin Secretion by Repressing PDX1 Transcriptional Actvity and Is Down-regulated in Hyperinsulinemic Obese Mice
J. Biol. Chem., November 11, 2005; 280(45): 37669 - 37680.
[Abstract] [Full Text] [PDF]

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 Am. J. Physiol. Endocrinol. Metab.Home page
J. T. Taylor, L. Huang, B. M. Keyser, H. Zhuang, C. W. Clarkson, and M. Li
Role of high-voltage-activated calcium channels in glucose-regulated {beta}-cell calcium homeostasis and insulin release
Am J Physiol Endocrinol Metab, November 1, 2005; 289(5): E900 - E908.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
J. Rasschaert, L. Ladriere, M. Urbain, Z. Dogusan, B. Katabua, S. Sato, S. Akira, C. Gysemans, C. Mathieu, and D. L. Eizirik
Toll-like Receptor 3 and STAT-1 Contribute to Double-stranded RNA+ Interferon-{gamma}-induced Apoptosis in Primary Pancreatic {beta}-Cells
J. Biol. Chem., October 7, 2005; 280(40): 33984 - 33991.
[Abstract] [Full Text] [PDF]

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 J EndocrinolHome page
J. Jensen, E. D Galsgaard, A. E Karlsen, Y. C Lee, and J. H Nielsen
STAT5 activation by human GH protects insulin-producing cells against interleukin-1{beta}, interferon-{gamma} and tumour necrosis factor-{alpha}-induced apoptosis independent of nitric oxide production
J. Endocrinol., October 1, 2005; 187(1): 25 - 36.
[Abstract] [Full Text] [PDF]

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 J. Cell Sci.Home page
S. Obermuller, A. Lindqvist, J. Karanauskaite, J. Galvanovskis, P. Rorsman, and S. Barg
Selective nucleotide-release from dense-core granules in insulin-secreting cells
J. Cell Sci., September 15, 2005; 118(18): 4271 - 4282.
[Abstract] [Full Text] [PDF]

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 DiabetesHome page
R. Veluthakal, I. Chvyrkova, M. Tannous, P. McDonald, R. Amin, T. Hadden, D. C. Thurmond, M. J. Quon, and A. Kowluru
Essential Role for Membrane Lipid Rafts in Interleukin-1{beta}-Induced Nitric Oxide Release From Insulin-Secreting Cells: Potential Regulation by Caveolin-1+
Diabetes, September 1, 2005; 54(9): 2576 - 2585.
[Abstract] [Full Text] [PDF]

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 J. Cell Sci.Home page
H. Wang, G. Kouri, and C. B. Wollheim
ER stress and SREBP-1 activation are implicated in {beta}-cell glucolipotoxicity
J. Cell Sci., September 1, 2005; 118(17): 3905 - 3915.
[Abstract] [Full Text] [PDF]

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 J EndocrinolHome page
C. W Hay, E. M Sinclair, G. Bermano, E. Durward, M. Tadayyon, and K. Docherty
Glucagon-like peptide-1 stimulates human insulin promoter activity in part through cAMP-responsive elements that lie upstream and downstream of the transcription start site
J. Endocrinol., August 1, 2005; 186(2): 353 - 365.
[Abstract] [Full Text] [PDF]

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 Mol. Cell. Biol.Home page
D. Lu, T. Ventura-Holman, J. Li, R. W. McMurray, J. S. Subauste, and J. F. Maher
Abnormal Glucose Homeostasis and Pancreatic Islet Function in Mice with Inactivation of the Fem1b Gene
Mol. Cell. Biol., August 1, 2005; 25(15): 6570 - 6577.
[Abstract] [Full Text] [PDF]

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 Mol. Endocrinol.Home page
L. Marzban, G. Trigo-Gonzalez, and C. B. Verchere
Processing of Pro-Islet Amyloid Polypeptide in the Constitutive and Regulated Secretory Pathways of {beta} Cells
Mol. Endocrinol., August 1, 2005; 19(8): 2154 - 2163.
[Abstract] [Full Text] [PDF]

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 J EndocrinolHome page
M Asfari, S Berta, S Coffy, M Kergoat, C Charon, and A Audet
Short-term treatment with mycophenolic acid and tacrolimus is tolerogenic for INS-1 cell clone transplantation and the deleterious effects of the drugs are limited: in vivo and in vitro studies
J. Endocrinol., July 1, 2005; 186(1): 213 - 220.
[Abstract] [Full Text] [PDF]

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 DiabetesHome page
K. N. Samli, M. J. McGuire, C. B. Newgard, S. A. Johnston, and K. C. Brown
Peptide-Mediated Targeting of the Islets of Langerhans
Diabetes, July 1, 2005; 54(7): 2103 - 2108.
[Abstract] [Full Text] [PDF]

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 J. Physiol.Home page
G. Kang, O. G Chepurny, M. J Rindler, L. Collis, Z. Chepurny, W.-h. Li, M. Harbeck, M. W Roe, and G. G Holz
A cAMP and Ca2+ coincidence detector in support of Ca2+-induced Ca2+ release in mouse pancreatic {beta} cells
J. Physiol., July 1, 2005; 566(1): 173 - 188.
[Abstract] [Full Text] [PDF]

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 Proc. Natl. Acad. Sci. USAHome page
J. C. Schisler, P. B. Jensen, D. G. Taylor, T. C. Becker, F. K. Knop, S. Takekawa, M. German, G. C. Weir, D. Lu, R. G. Mirmira, et al.
The Nkx6.1 homeodomain transcription factor suppresses glucagon expression and regulates glucose-stimulated insulin secretion in islet beta cells
PNAS, May 17, 2005; 102(20): 7297 - 7302.
[Abstract] [Full Text] [PDF]

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 Mol. Endocrinol.Home page
M. F. Pino, D. Z. Ye, K. D. Linning, C. D. Green, B. Wicksteed, V. Poitout, and L. K. Olson
Elevated Glucose Attenuates Human Insulin Gene Promoter Activity in INS-1 Pancreatic {beta}-Cells via Reduced Nuclear Factor Binding to the A5/Core and Z Element
Mol. Endocrinol., May 1, 2005; 19(5): 1343 - 1360.
[Abstract] [Full Text] [PDF]

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 DiabetesHome page
L. D. Mastrandrea, S. M. Sessanna, and S. G. Laychock
Sphingosine Kinase Activity and Sphingosine-1 Phosphate Production in Rat Pancreatic Islets and INS-1 Cells: Response to Cytokines
Diabetes, May 1, 2005; 54(5): 1429 - 1436.
[Abstract] [Full Text] [PDF]

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 DiabetesHome page
D. E. Johnson, H. Yamazaki, K. M. Ward, A. W. Schmidt, W. S. Lebel, J. L. Treadway, E. M. Gibbs, W. S. Zawalich, and H. Rollema
Inhibitory Effects of Antipsychotics on Carbachol-Enhanced Insulin Secretion From Perifused Rat Islets: Role of Muscarinic Antagonism in Antipsychotic-Induced Diabetes and Hyperglycemia
Diabetes, May 1, 2005; 54(5): 1552 - 1558.
[Abstract] [Full Text] [PDF]

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 DiabetesHome page
S. Ullrich, S. Berchtold, F. Ranta, G. Seebohm, G. Henke, A. Lupescu, A. F. Mack, C.-M. Chao, J. Su, R. Nitschke, et al.
Serum- and Glucocorticoid-Inducible Kinase 1 (SGK1) Mediates Glucocorticoid-Induced Inhibition of Insulin Secretion
Diabetes, April 1, 2005; 54(4): 1090 - 1099.
[Abstract] [Full Text] [PDF]

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 EndocrinologyHome page
C. A. Gysemans, A. K. Cardozo, H. Callewaert, A. Giulietti, L. Hulshagen, R. Bouillon, D. L. Eizirik, and C. Mathieu
1,25-Dihydroxyvitamin D3 Modulates Expression of Chemokines and Cytokines in Pancreatic Islets: Implications for Prevention of Diabetes in Nonobese Diabetic Mice
Endocrinology, April 1, 2005; 146(4): 1956 - 1964.
[Abstract] [Full Text] [PDF]

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 DiabetesHome page
P. E. MacDonald, S. Obermuller, J. Vikman, J. Galvanovskis, P. Rorsman, and L. Eliasson
Regulated Exocytosis and Kiss-and-Run of Synaptic-Like Microvesicles in INS-1 and Primary Rat {beta}-Cells
Diabetes, March 1, 2005; 54(3): 736 - 743.
[Abstract] [Full Text] [PDF]

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 DiabetesHome page
L. Herrero, B. Rubi, D. Sebastian, D. Serra, G. Asins, P. Maechler, M. Prentki, and F. G. Hegardt
Alteration of the Malonyl-CoA/Carnitine Palmitoyltransferase I Interaction in the {beta}-Cell Impairs Glucose-Induced Insulin Secretion
Diabetes, February 1, 2005; 54(2): 462 - 471.
[Abstract] [Full Text] [PDF]

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 EndocrinologyHome page
C. Longuet, C. Broca, S. Costes, E. H. Hani, D. Bataille, and S. Dalle
Extracellularly Regulated Kinases 1/2 (p44/42 Mitogen-Activated Protein Kinases) Phosphorylate Synapsin I and Regulate Insulin Secretion in the MIN6 {beta}-Cell Line and Islets of Langerhans
Endocrinology, February 1, 2005; 146(2): 643 - 654.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
I. Briaud, L. M. Dickson, M. K. Lingohr, J. F. McCuaig, J. C. Lawrence, and C. J. Rhodes
Insulin Receptor Substrate-2 Proteasomal Degradation Mediated by a Mammalian Target of Rapamycin (mTOR)-induced Negative Feedback Down-regulates Protein Kinase B-mediated Signaling Pathway in {beta}-Cells
J. Biol. Chem., January 21, 2005; 280(3): 2282 - 2293.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
B. Rubi, A. del Arco, C. Bartley, J. Satrustegui, and P. Maechler
The Malate-Aspartate NADH Shuttle Member Aralar1 Determines Glucose Metabolic Fate, Mitochondrial Activity, and Insulin Secretion in Beta Cells
J. Biol. Chem., December 31, 2004; 279(53): 55659 - 55666.
[Abstract] [Full Text] [PDF]

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 JCBHome page
M. Trajkovski, H. Mziaut, A. Altkruger, J. Ouwendijk, K.-P. Knoch, S. Muller, and M. Solimena
Nuclear translocation of an ICA512 cytosolic fragment couples granule exocytosis and insulin expression in {beta}-cells
J. Cell Biol., December 20, 2004; 167(6): 1063 - 1074.
[Abstract] [Full Text] [PDF]


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