In vivo differential effects of fasting, re-feeding, insulin and insulin stimulation time course on insulin signaling pathway components in peripheral tissues

Abdelali Agouni, Carl Owen, Alicja Czopek, Nimesh Mody (Corresponding Author), Mirela Delibegovic (Corresponding Author)

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Abstract

Background: Components of the insulin receptor signaling pathway are probably
some of the best studied ones. Even though methods for studying these components are well established, the in vivo effects of different fasting regimens, and the time-course of insulin receptor phosphorylation and that of its downstream components in insulinsensitive peripheral tissues have not been analyzed in detail. Rationale: When assessing insulin signalling, it may be beneficial to drive insulin levels as low as possible by performing an overnight fast before injecting a supra-physiological dose of insulin. Recent studies have shown however that 5 or 6 hour fast in mice is sufficient to assess physiological responses to insulin and/or glucose in glucose tolerance tests (1), insulin tolerance tests (2-4) and euglycemic hyperinsulinemic clamp studies (5, 6). Moreover, mice are nocturnal feeders, with ~70% of their daily caloric intake occurring during the dark cycle (5), and their metabolic rate is much higher than humans. Therefore, an overnight fast in mice is closer to starvation than just food withdrawal. Thus our aim was *Manuscript Click here to view linked References
to assess insulin signalling components from the insulin receptor to downstream targets IRS1, Akt/PKB, GSK3, Erk1/2 and ribosomal protein S6 in muscle, liver and adipose tissue in 5 hour versus 16 hour (overnight) fasted mice, and the time-course of these phosphorylation events. We also assessed whether re-feeding under 5 hour and 16 hour fasting conditions was a more robust stimulus than insulin alone. Conclusions: Our study determines that a short food withdrawal from mice, for a period of 5 hours, results in a similar insulin-stimulated response in phosphorylation events as the long overnight fast, presenting a more physiological experimental set up. We also demonstrate that in vivo, insulin stimulated phosphorylation of its signaling components is different between different peripheral tissues, and depending on the tissue(s) and protein(s) of interest, an appropriate time-course should be chosen.
Original languageEnglish
Pages (from-to)104-111
Number of pages8
JournalBiochemical and Biophysical Research Communications
Volume401
Issue number1
Early online date15 Sep 2010
DOIs
Publication statusPublished - 8 Oct 2010

Fingerprint

Fasting
Insulin
Tissue
Phosphorylation
Insulin Receptor
Ribosomal Protein S6
Glucose
Food
Glucose Clamp Technique
Manuscripts
Clamping devices
Glucose Tolerance Test
Starvation
Energy Intake
Liver
Muscle
Adipose Tissue
Muscles
Proteins

Keywords

  • insulin signalling
  • phosphorylation
  • fasting
  • re-feeding
  • insulin

Cite this

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title = "In vivo differential effects of fasting, re-feeding, insulin and insulin stimulation time course on insulin signaling pathway components in peripheral tissues",
abstract = "Background: Components of the insulin receptor signaling pathway are probably some of the best studied ones. Even though methods for studying these components are well established, the in vivo effects of different fasting regimens, and the time-course of insulin receptor phosphorylation and that of its downstream components in insulinsensitive peripheral tissues have not been analyzed in detail. Rationale: When assessing insulin signalling, it may be beneficial to drive insulin levels as low as possible by performing an overnight fast before injecting a supra-physiological dose of insulin. Recent studies have shown however that 5 or 6 hour fast in mice is sufficient to assess physiological responses to insulin and/or glucose in glucose tolerance tests (1), insulin tolerance tests (2-4) and euglycemic hyperinsulinemic clamp studies (5, 6). Moreover, mice are nocturnal feeders, with ~70{\%} of their daily caloric intake occurring during the dark cycle (5), and their metabolic rate is much higher than humans. Therefore, an overnight fast in mice is closer to starvation than just food withdrawal. Thus our aim was *Manuscript Click here to view linked References to assess insulin signalling components from the insulin receptor to downstream targets IRS1, Akt/PKB, GSK3, Erk1/2 and ribosomal protein S6 in muscle, liver and adipose tissue in 5 hour versus 16 hour (overnight) fasted mice, and the time-course of these phosphorylation events. We also assessed whether re-feeding under 5 hour and 16 hour fasting conditions was a more robust stimulus than insulin alone. Conclusions: Our study determines that a short food withdrawal from mice, for a period of 5 hours, results in a similar insulin-stimulated response in phosphorylation events as the long overnight fast, presenting a more physiological experimental set up. We also demonstrate that in vivo, insulin stimulated phosphorylation of its signaling components is different between different peripheral tissues, and depending on the tissue(s) and protein(s) of interest, an appropriate time-course should be chosen.",
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T1 - In vivo differential effects of fasting, re-feeding, insulin and insulin stimulation time course on insulin signaling pathway components in peripheral tissues

AU - Agouni, Abdelali

AU - Owen, Carl

AU - Czopek, Alicja

AU - Mody, Nimesh

AU - Delibegovic, Mirela

PY - 2010/10/8

Y1 - 2010/10/8

N2 - Background: Components of the insulin receptor signaling pathway are probably some of the best studied ones. Even though methods for studying these components are well established, the in vivo effects of different fasting regimens, and the time-course of insulin receptor phosphorylation and that of its downstream components in insulinsensitive peripheral tissues have not been analyzed in detail. Rationale: When assessing insulin signalling, it may be beneficial to drive insulin levels as low as possible by performing an overnight fast before injecting a supra-physiological dose of insulin. Recent studies have shown however that 5 or 6 hour fast in mice is sufficient to assess physiological responses to insulin and/or glucose in glucose tolerance tests (1), insulin tolerance tests (2-4) and euglycemic hyperinsulinemic clamp studies (5, 6). Moreover, mice are nocturnal feeders, with ~70% of their daily caloric intake occurring during the dark cycle (5), and their metabolic rate is much higher than humans. Therefore, an overnight fast in mice is closer to starvation than just food withdrawal. Thus our aim was *Manuscript Click here to view linked References to assess insulin signalling components from the insulin receptor to downstream targets IRS1, Akt/PKB, GSK3, Erk1/2 and ribosomal protein S6 in muscle, liver and adipose tissue in 5 hour versus 16 hour (overnight) fasted mice, and the time-course of these phosphorylation events. We also assessed whether re-feeding under 5 hour and 16 hour fasting conditions was a more robust stimulus than insulin alone. Conclusions: Our study determines that a short food withdrawal from mice, for a period of 5 hours, results in a similar insulin-stimulated response in phosphorylation events as the long overnight fast, presenting a more physiological experimental set up. We also demonstrate that in vivo, insulin stimulated phosphorylation of its signaling components is different between different peripheral tissues, and depending on the tissue(s) and protein(s) of interest, an appropriate time-course should be chosen.

AB - Background: Components of the insulin receptor signaling pathway are probably some of the best studied ones. Even though methods for studying these components are well established, the in vivo effects of different fasting regimens, and the time-course of insulin receptor phosphorylation and that of its downstream components in insulinsensitive peripheral tissues have not been analyzed in detail. Rationale: When assessing insulin signalling, it may be beneficial to drive insulin levels as low as possible by performing an overnight fast before injecting a supra-physiological dose of insulin. Recent studies have shown however that 5 or 6 hour fast in mice is sufficient to assess physiological responses to insulin and/or glucose in glucose tolerance tests (1), insulin tolerance tests (2-4) and euglycemic hyperinsulinemic clamp studies (5, 6). Moreover, mice are nocturnal feeders, with ~70% of their daily caloric intake occurring during the dark cycle (5), and their metabolic rate is much higher than humans. Therefore, an overnight fast in mice is closer to starvation than just food withdrawal. Thus our aim was *Manuscript Click here to view linked References to assess insulin signalling components from the insulin receptor to downstream targets IRS1, Akt/PKB, GSK3, Erk1/2 and ribosomal protein S6 in muscle, liver and adipose tissue in 5 hour versus 16 hour (overnight) fasted mice, and the time-course of these phosphorylation events. We also assessed whether re-feeding under 5 hour and 16 hour fasting conditions was a more robust stimulus than insulin alone. Conclusions: Our study determines that a short food withdrawal from mice, for a period of 5 hours, results in a similar insulin-stimulated response in phosphorylation events as the long overnight fast, presenting a more physiological experimental set up. We also demonstrate that in vivo, insulin stimulated phosphorylation of its signaling components is different between different peripheral tissues, and depending on the tissue(s) and protein(s) of interest, an appropriate time-course should be chosen.

KW - insulin signalling

KW - phosphorylation

KW - fasting

KW - re-feeding

KW - insulin

U2 - 10.1016/j.bbrc.2010.09.018

DO - 10.1016/j.bbrc.2010.09.018

M3 - Article

VL - 401

SP - 104

EP - 111

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 0006-291X

IS - 1

ER -