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Role of Akt isoforms in neuronal insulin signaling and resistance

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Abstract

The aim of the present study was to determine the role of Akt isoforms in insulin signaling and resistance in neuronal cells. By silencing Akt isoforms individually and in pairs, in Neuro-2a and HT22 cells we observed that, in insulin-sensitive condition, Akt isoforms differentially reduced activation of AS160 and glucose uptake with Akt2 playing the major role. Under insulin-resistant condition, phosphorylation of all isoforms and glucose uptake were severely affected. Over-expression of individual isoforms in insulin-sensitive and resistant cells differentially reversed AS160 phosphorylation with concomitant reversal in glucose uptake indicating a compensatory role of Akt isoforms in controlling neuronal insulin signaling. Post-insulin stimulation Akt2 translocated to the membrane the most followed by Akt3 and Akt1, decreasing glucose uptake in the similar order in insulin-sensitive cells. None of the Akt isoforms translocated in insulin-resistant cells or high-fat-diet mediated diabetic mice brain cells. Based on our data, insulin-dependent differential translocation of Akt isoforms to the plasma membrane turns out to be the key factor in determining Akt isoform specificity. Thus, isoforms play parallel with predominant role by Akt2, and compensatory yet novel role by Akt1 and Akt3 to regulate neuronal insulin signaling, glucose uptake, and insulin-resistance.

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Acknowledgements

We are grateful to Indian Institute of Technology-Delhi for their support. We would like to thank Dr. Prosenjit Mondal, Indian Institute of Technology—Mandi, Himachal Pradesh, India, for helping us in providing with normal diet and high-fat-fed diet mice whole brain samples. We would like to thank Dr. Nishi Raj, Jamia Hamdard-Institute of Molecular Medicine (JHIMM), New Delhi, India for providing pCMV6 Empty vector backbone. We would like to thank Dr. Chinmoy Mukhopadhyay, Jawaharalal Nehru University, New Delhi, for providing us with Caveolin-1 antibody. We would like to thank Dr. Pallavi Varshney for her assistance in training some methodologies. We are grateful to Advanced Technology Platform Centre, Regional Centre for Biotechnology, Faridabad, Haryana, India, for Confocal Microscopy.

Funding

MS is a recipient of Senior Research Fellowship (09/086 (1256)/2016-EMR-1) from Council of Scientific and Industrial Research (CSIR), Government of India. CSD is supported by a grant from Department of Biotechnology, Government of India, New Delhi, India (PR32795/MED/122/229/2019).

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  1. Kusuma School of Biological Sciences, Indian Institute of Technology-Delhi, Hauz Khas, New Delhi, 110016, India

    Medha Sharma & Chinmoy Sankar Dey

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  1. Medha Sharma
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MS performed all the experiments, analyzed the data and wrote the original drafts, approved the final version of the manuscript. CSD conceived the idea, provided resources, acquired the funding, written, reviewed and edited the manuscript and approved the final version of the manuscript.

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Correspondence to Chinmoy Sankar Dey.

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Mice whole brain tissues were kind gift from Dr. Prosenjit Mondal, Indian Institute of Technology—Mandi, HP, India. All experiments were performed following the guidelines prescribed by CPCSEA (Committee for the Purpose of Control and Supervision of Experiments on Animals) with the approval of the Internal Animal Ethics Committee, Visva-Bharati (IAEC/VB/2017/01). The legal requirements/guidelines in the country for the care and use of animals have been followed.

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Sharma, M., Dey, C.S. Role of Akt isoforms in neuronal insulin signaling and resistance. Cell. Mol. Life Sci. 78, 7873–7898 (2021). https://doi.org/10.1007/s00018-021-03993-6

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