Mallahalli S. Manu, Kuruvanthe S. Rachana, Gopal M. Advirao

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Presentation transcript:

Mallahalli S. Manu, Kuruvanthe S. Rachana, Gopal M. Advirao Altered expression of IRS2 and GRB2 in demyelination of peripheral neurons: Implications in diabetic neuropathy Mallahalli S. Manu, Kuruvanthe S. Rachana, Gopal M. Advirao Neuropeptides Volume 62, Pages 71-79 (April 2017) DOI: 10.1016/j.npep.2016.12.004 Copyright © 2016 Elsevier Ltd Terms and Conditions

Fig. 1 Alteration in the blood glucose levels of normal control, STZ and insulin treated STZ rats. Error bars represent±SEM (n=6). Neuropeptides 2017 62, 71-79DOI: (10.1016/j.npep.2016.12.004) Copyright © 2016 Elsevier Ltd Terms and Conditions

Fig. 2 Histological examination of tolidine blue stained longitudinal section of sciatic nerve showing the structural difference between 4, 8 and 12week sample of normal[(control) A, B, C] diabetic[(STZ) D, E, F] and insulin treated diabetic samples[(STZ+I) G, H, I]. Arrow heads showing deformation in nerve fiber organization. Scale bar: 50μm. Neuropeptides 2017 62, 71-79DOI: (10.1016/j.npep.2016.12.004) Copyright © 2016 Elsevier Ltd Terms and Conditions

Fig. 3 Differential expression of pIRS1 protein and IRS1 mRNA level at 4, 8 and 12week sciatic nerve samples correlated between control (C), diabetic (STZ) and insulin treated (STZ+I) and 12h and 24h of the SCs grown in normal medium (C), 40mM glucose (H-GLU) and 10nM insulin added 40mM glucose medium (H-GLU+I). (A and B) Western blot analysis showing relative fold change in pIRS1 protein expression of sciatic nerve in STZ and STZ+I compared to C. (C and D) Western blot analysis showing relative fold change in pIRS1 protein expression of SCs in H-GLU and H-GLU+I compared to C. (E) Real time PCR studies showing change in relative mRNA expression among STZ and STZ+I of sciatic nerve. (F) Real time PCR studies showing change in relative mRNA expression among H-GLU and H-GLU+I of SCs. * indicates p<0.05. Neuropeptides 2017 62, 71-79DOI: (10.1016/j.npep.2016.12.004) Copyright © 2016 Elsevier Ltd Terms and Conditions

Fig. 4 Immunofluorescence staining for evaluation of pIRS1 protein expression difference among SCs grown for 12h and 24h in normal medium[(control)A and B], high glucose medium[(40mM glucose) C and D] and insulin added high glucose medium[(40mM glucose+10nM insulin) E and F]. Scale bar: 50μm. Neuropeptides 2017 62, 71-79DOI: (10.1016/j.npep.2016.12.004) Copyright © 2016 Elsevier Ltd Terms and Conditions

Fig. 5 Relative mRNA expression of IRS2 (A) IRS2 mRNA expression of 4, 8 and 12week sciatic nerve of control (C), diabetic (STZ) and insulin treated diabetic rats (STZ+I). (B) Relative mRNA expression of IRS2 among SCs grown in 40mM glucose (H-GLU) and 10nM insulin added 40mM glucose medium (H-GLU+I) at 12h and 24h time interval * indicates p<0.05. Neuropeptides 2017 62, 71-79DOI: (10.1016/j.npep.2016.12.004) Copyright © 2016 Elsevier Ltd Terms and Conditions

Fig. 6 Western blot analysis of pS(731)IRS2 in sciatic nerve and SC. (A and B) Expression of pS(731)IRS2 protein of 4, 8 and 12week sciatic nerve samples correlated between control (C), diabetic (STZ) and insulin treated diabetic rats (STZ+I). (B and C) Relative fold change of pS(731)IRS2 at 12h and 24h of the SCs grown in normal medium (C), 40mM glucose (H-GLU) and 10nM insulin added 40mM glucose medium (H-GLU+I). * indicates p<0.05. Neuropeptides 2017 62, 71-79DOI: (10.1016/j.npep.2016.12.004) Copyright © 2016 Elsevier Ltd Terms and Conditions

Fig. 7 Immunofluorescence staining for evaluation of pS(731)IRS2 protein expression difference among SCs grown for 12h and 24h in normal medium [(control) A and B], high glucose medium [(40mM glucose) C and D] and insulin added high glucose medium [(40mM glucose+10nM insulin) E and F]. Scale bar: 50μm. Neuropeptides 2017 62, 71-79DOI: (10.1016/j.npep.2016.12.004) Copyright © 2016 Elsevier Ltd Terms and Conditions

Fig. 8 Differential expression of pGRB2 protein and GRB2 mRNA level at 4, 8 and 12week sciatic nerve samples correlated between control (C), diabetic (STZ) and insulin treated diabetic rats (STZ+I). 12h and 24h of the SCs grown in normal medium (C), 40mM glucose (H-GLU) and 10nM insulin added 40mM glucose medium (H-GLU+I). (A and B) Western blot analysis showing relative fold change in pGRB2 protein expression of sciatic nerve in STZ and STZ+I compared to C. (C and D) Western blot analysis showing relative fold change in pGRB2 protein expression of SCs in H-GLU and H-GLU+I compared to C. (E) Real time PCR studies showing change in relative mRNA expression of GRB2 among STZ and STZ+I of sciatic nerve. (F) Real time PCR studies showing change in relative mRNA expression of GRB2 among H-GLU and H-GLU+I of SCs. * indicates p<0.05. Neuropeptides 2017 62, 71-79DOI: (10.1016/j.npep.2016.12.004) Copyright © 2016 Elsevier Ltd Terms and Conditions

Fig. 9 Immunofluorescence staining for evaluation of pGRB2 protein expression difference among SCs grown for 12h and 24h in normal medium [(control) A and B], high glucose medium [(40mM glucose) C and D] and insulin added high glucose medium [(40mM glucose+10nM insulin) E and F]. Scale bar: 50μm. Neuropeptides 2017 62, 71-79DOI: (10.1016/j.npep.2016.12.004) Copyright © 2016 Elsevier Ltd Terms and Conditions

Neuropeptides 2017 62, 71-79DOI: (10.1016/j.npep.2016.12.004) Copyright © 2016 Elsevier Ltd Terms and Conditions