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The flower pot method of REM sleep deprivation causes apoptotic cell death in the hepatocytes of rat

View ORCID ProfileAtul Pandey, Devesh Kumar, Gopesh Ray, Santosh Kar
doi: https://doi.org/10.1101/375717
Atul Pandey
1School of Biotechnology, Jawaharlal Nehru University, New Delhi-110067, India
2Department of Ecology, Evolution, and Behavior, The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem, 91904, Israel
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  • For correspondence: atulkuma.pandey@mail.huji.ac.il santoshkariis@rediffmail.com
Devesh Kumar
1School of Biotechnology, Jawaharlal Nehru University, New Delhi-110067, India
3University Hospital Brussels, Brussels, Belgium
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Gopesh Ray
1School of Biotechnology, Jawaharlal Nehru University, New Delhi-110067, India
4School of Biotechnology, KIIT University, Bhubaneswar-751024, Odisha, India
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Santosh Kar
1School of Biotechnology, Jawaharlal Nehru University, New Delhi-110067, India
4School of Biotechnology, KIIT University, Bhubaneswar-751024, Odisha, India
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  • For correspondence: atulkuma.pandey@mail.huji.ac.il santoshkariis@rediffmail.com
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Abstract

Introduction The rapid eye movement sleep deprivation (REMSD) of rats relates with increased inflammations, acute phase response, oxidative damage, neuronal cell loss, and neurodegenerative diseases. Whereas, its role outside brain are not well studied. This study tried to explore the causal effect of REM sleep loss on hepatocytes.

Methods We deprived the rats of REM sleep using standard flower pot method. We focused on liver to see the REMSD affects which controls most of the metabolic processes of the body.

Results We report here that flower pot induced REMSD causes apoptotic cell death of hepatocytes (~10% by Annexin Assay & ~20% by TUNEL assay). This were further got alleviated up to extent after sleep recovery of 5 days (recovered approximately 8.0% by Annexin Assay & 14% by TUNEL assay). The gene expression and protein level profiling revealed the up-regulation of p53, Bax, Cytochrome c, Caspase 3, and Caspase 9. While, Bcl2 which is an anti-apoptotic protein were down-regulated in response to REMSD. Relentless recovery of 5 days affected the expression pattern of these genes/proteins.

Conclusions Our study offer great pathological and physiological significance for sleep loss, by inferring the apoptotic cell-death in the hepatocytes of rat. This further signifies the functional and preventive role of REM sleep which is unique to mammals and avians with certain exceptions, as its loss can affect the natural well-being and survival of the individuals.

Highlights of the study

  • We observed significant apoptosis in the hepatocytes of REMSD group of rats.

  • Our expression analysis confirmed altered expression for genes p53, Bcl2, Bax, and Caspase-3 after REMSD.

  • Protein level analysis supported our gene expression results for p53, Bcl2, Bax, Caspase 3 and Caspase 9 after REMSD.

  • Sleep recovery improved the respective genes and protein expression levels towards normalcy, signifying the functional role of REM sleep.

  • Abbreviations

    REM
    rapid eye movement
    NREM
    non rapid eye movement
    REMSD
    rapid eye movement sleep deprivationn
    CC
    cage control
    LPC
    large platform control
    TUNEL
    Terminal deoxynucleotidyl transferase dUTP nick end labeling
    EDTA
    Ethylene diamine tetra acetic acid
    RIPA
    Radio immuno precipitation assay
    FITC
    Fluorescein isothiocyanate
    TDT
    Terminal deoxynucleotidyl transferase
    FAM
    6-carboxyfluorescein
    ROS
    Reactive oxygen species
    NO
    Nitric Oxide
  • Copyright 
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    Posted October 18, 2019.
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    The flower pot method of REM sleep deprivation causes apoptotic cell death in the hepatocytes of rat
    Atul Pandey, Devesh Kumar, Gopesh Ray, Santosh Kar
    bioRxiv 375717; doi: https://doi.org/10.1101/375717
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    The flower pot method of REM sleep deprivation causes apoptotic cell death in the hepatocytes of rat
    Atul Pandey, Devesh Kumar, Gopesh Ray, Santosh Kar
    bioRxiv 375717; doi: https://doi.org/10.1101/375717

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