Insulin Rescues Retinal Neurons from Apoptosis by a Phosphatidylinositol 3-Kinase/Akt-mediated Mechanism That Reduces the Activation of Caspase-3

Alistair Barber, Makoto Nakamura, Ellen B. Wolpert, Chad E.N. Reiter, Gail M. Seigel, David A. Antonetti, Thomas W. Gardner

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Abstract

The ability of insulin to protect neurons from apoptosis was examined in differentiated R28 cells, a neural cell line derived from the neonatal rat retina. Apoptosis was induced by serum deprivation, and the number of pyknotic cells was counted. p53 and Akt were examined by immunoblotting after serum deprivation and insulin treatment, and caspase-3 activation was examined by immunocytochemistry. Serum deprivation for 24 h caused ∼20% of R28 cells to undergo apoptosis, detected by both pyknosis and activation of caspase-3. 10 nM insulin maximally reduced the amount of apoptosis with a similar potency as 1.3 nM (10 ng/ml) insulin-like growth factor 1, which acted as a positive control. Insulin induced serine phosphorylation of Akt, through the phosphatidylinositol (PI) 3-kinase pathway. Inhibition of PI 3-kinase with wortmannin or LY294002 blocked the ability of insulin to rescue the cells from apoptosis. SN50, a peptide inhibitor of NF-κB nuclear translocation, blocked the rescue effect of insulin, but neither insulin or serum deprivation induced phosphorylation of IκB. These results suggest that insulin is a survival factor for retinal neurons by activating the PI 3-kinase/Akt pathway and by reducing caspase-3 activation. The rescue effect of insulin does not appear to be mediated by NF-κB or p53. These data suggest that insulin provides trophic support for retinal neurons through a PI 3-kinase/Akt-dependent pathway.

Original languageEnglish (US)
Pages (from-to)32814-32821
Number of pages8
JournalJournal of Biological Chemistry
Volume276
Issue number35
DOIs
StatePublished - Aug 31 2001

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Phosphatidylinositol 3-Kinase
Retinal Neurons
Caspase 3
Neurons
Chemical activation
Insulin
Apoptosis
Phosphorylation
Serum
2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
Somatomedins
Immunoblotting
Serine
Retina
Rats
Cell Count
Immunohistochemistry
Cells

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Barber, Alistair ; Nakamura, Makoto ; Wolpert, Ellen B. ; Reiter, Chad E.N. ; Seigel, Gail M. ; Antonetti, David A. ; Gardner, Thomas W. / Insulin Rescues Retinal Neurons from Apoptosis by a Phosphatidylinositol 3-Kinase/Akt-mediated Mechanism That Reduces the Activation of Caspase-3. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 35. pp. 32814-32821.
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abstract = "The ability of insulin to protect neurons from apoptosis was examined in differentiated R28 cells, a neural cell line derived from the neonatal rat retina. Apoptosis was induced by serum deprivation, and the number of pyknotic cells was counted. p53 and Akt were examined by immunoblotting after serum deprivation and insulin treatment, and caspase-3 activation was examined by immunocytochemistry. Serum deprivation for 24 h caused ∼20{\%} of R28 cells to undergo apoptosis, detected by both pyknosis and activation of caspase-3. 10 nM insulin maximally reduced the amount of apoptosis with a similar potency as 1.3 nM (10 ng/ml) insulin-like growth factor 1, which acted as a positive control. Insulin induced serine phosphorylation of Akt, through the phosphatidylinositol (PI) 3-kinase pathway. Inhibition of PI 3-kinase with wortmannin or LY294002 blocked the ability of insulin to rescue the cells from apoptosis. SN50, a peptide inhibitor of NF-κB nuclear translocation, blocked the rescue effect of insulin, but neither insulin or serum deprivation induced phosphorylation of IκB. These results suggest that insulin is a survival factor for retinal neurons by activating the PI 3-kinase/Akt pathway and by reducing caspase-3 activation. The rescue effect of insulin does not appear to be mediated by NF-κB or p53. These data suggest that insulin provides trophic support for retinal neurons through a PI 3-kinase/Akt-dependent pathway.",
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Insulin Rescues Retinal Neurons from Apoptosis by a Phosphatidylinositol 3-Kinase/Akt-mediated Mechanism That Reduces the Activation of Caspase-3. / Barber, Alistair; Nakamura, Makoto; Wolpert, Ellen B.; Reiter, Chad E.N.; Seigel, Gail M.; Antonetti, David A.; Gardner, Thomas W.

In: Journal of Biological Chemistry, Vol. 276, No. 35, 31.08.2001, p. 32814-32821.

Research output: Contribution to journalArticle

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T1 - Insulin Rescues Retinal Neurons from Apoptosis by a Phosphatidylinositol 3-Kinase/Akt-mediated Mechanism That Reduces the Activation of Caspase-3

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