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Molecular Basis of Sex Difference in Neuroprotection induced by Hypoxia Preconditioning in Zebrafish

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Abstract

Hypoxia, the major cause of ischemic injury, leads to debilitating disease in infants via birth asphyxia and cerebral palsy, whereas in adults via heart attack and stroke. A widespread, natural protective phenomenon termed ‘hypoxic preconditioning’ (PH) occurs when prior exposures to hypoxia eventually result in robust hypoxia resistance. Accordingly, we have developed and optimized a novel model of hypoxic preconditioning in adult zebrafish to mimic the tolerance of mini stroke(s) in human, which appears to protect against the severe damage inflicted by a major stroke event. Here, we observed a remarkable difference in the progression pattern of neuroprotection between preconditioning hypoxia followed by acute hypoxia (PH) group, and acute hypoxia (AH) only group, with noticeable sex difference when compared with normoxia behaviour upon recovery. Since gender difference has been reported in stroke risk factors and disease history, it was pertinent to investigate whether any such sex difference also exists in PH’s protective mechanism against acute ischemic stroke. In order to elucidate the neural molecular mechanisms behind sex difference in neuroprotection induced by PH, a high throughput proteomics approach utilizing iTRAQ was performed, followed by protein enrichment analysis using ingenuity pathway analysis (IPA) tool. Out of thousands of significantly altered proteins in zebrafish brain, the ones having critical role either in neuroglial proliferation/differentiation or neurotrophic functions were validated by analyzing their expression levels in preconditioned (PH), acute hypoxia (AH), and normoxia groups. The data indicate that female zebrafish brains are more protected against the severity of AH when exposed to the hypoxic preconditioning. The study also sheds light on the involvement of many signalling pathways underlying sex difference in preconditioning-induced neuroprotective mechanism, which can be further validated for the therapeutic approach.

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Acknowledgements

We thank the Director, CSIR-IICT for overall support and KIM department for generating an institutional publication number (IICT/Pubs./2019/437).

Funding

This research was supported by the Council of Scientific and Industrial Research (CSIR), India network project (BSC0103-UNDO to SC and AK), and Department of Biotechnology, Government of India (BT/PR14338/MED/30/495/2010 to SC).

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Authors and Affiliations

  1. Applied Biology, CSIR-Indian Institute of Chemical Technology (IICT), Hyderabad, India

    Tapatee Das, Kalyani Soren, Mounica Yerasi & Sumana Chakravarty

  2. Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, U.P., 201002, India

    Tapatee Das, Kalyani Soren, Arvind Kumar & Sumana Chakravarty

  3. CSIR-Centre for Cellular and Molecular Biology (CCMB), Hyderabad, India

    Avijeet Kamle & Arvind Kumar

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Supplementary Fig.1

IPA generated protein enrichment analysis . Top 5 canonical pathways of PHM and PHF (A) . Top upstream regulators of PHM and PHF (B) (PNG 406 kb)

Supplementary Fig. 2

Top regulatory networks generated by IPA. Regulatory networks with disease functions of observation 1 (AHM/AHF vs NM/NF) and observation 2 (PHM/PHF vs NM/NF). (PNG 976 kb)

Supplementary Table.1

IPA generated top analysis ready molecules. (PNG 701 kb)

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Das, T., Soren, K., Yerasi, M. et al. Molecular Basis of Sex Difference in Neuroprotection induced by Hypoxia Preconditioning in Zebrafish. Mol Neurobiol 57, 5177–5192 (2020). https://doi.org/10.1007/s12035-020-02091-1

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