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Sex dependent alterations in mitochondrial electron transport chain proteins following neonatal rat cerebral hypoxic-ischemia

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Abstract

Males are more susceptible to brain mitochondrial bioenergetic dysfunction following neonatal cerebral hypoxic-ischemia (HI) than females. Mitochondrial biogenesis has been implicated in the cellular response to HI injury, but sex differences in biogenesis following HI have not been described. We tested the hypothesis that mitochondrial biogenesis or the expression of mitochondrial electron transport chain (ETC) proteins are differentially stimulated in the brains of 8 day old male and female rats one day following HI, and promoted by treatment with acetyl-L-carnitine (ALCAR). There were no sex differences in mitochondrial mass, as reflected by the ratio of mitochondrial to nuclear DNA (mtDNA/nDNA) and citrate synthase enzyme activity present one day following HI or sham surgery. There was an increase in mtDNA/nDNA, however, in the hypoxic and ischemic (ipsilateral) hemisphere after HI in both male and female brains at one day post-injury, which was suppressed by ALCAR. Citrate synthase activity was increased in the ipsilateral hemisphere of ALCAR treated male and female brain. Most importantly, the levels of representative mitochondrial proteins present in ETC complexes I, II and IV increased substantially one day following HI in female, but not male brain. This sex difference is consistent with the increase in the mitochondrial biogenesis-associated transcription factor NRF-2/GABPα following HI in females, in contrast to the decrease observed with males. In conclusion, the female sex-selective increase in ETC proteins following HI may at least partially explain the relative female resilience to mitochondrial respiratory impairment and neuronal death that occur after HI.

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References

  • Almeida A, Bates TE, Clark JB (1994) Development of mitochondrial respiratory-chain complexes in neonatal rat brain. Biochem Soc Trans 22:409S

    Article  CAS  Google Scholar 

  • Almeida A, Brooks KJ, Sammut I, Keelan J, Davey GP, Clark JB, Bates TE (1995) Postnatal development of the complexes of the electron transport chain in synaptic mitochondria from rat brain. Dev Neurosci 17:212–218

    Article  CAS  Google Scholar 

  • Bates TE, Almeida A, Heales SJ, Clark JB (1994) Postnatal development of the complexes of the electron transport chain in isolated rat brain mitochondria. Dev Neurosci 16:321–327

    Article  CAS  Google Scholar 

  • Bird MI, Munday LA, Saggerson ED, Clark JB (1985) Carnitine acyltransferase activities in rat brain mitochondria. Bimodal distribution, kinetic constants, regulation by malonyl-CoA and developmental pattern. Biochem J 226:323–330

    Article  CAS  Google Scholar 

  • Demarest TG, Schuh RA, Waddell J, McKenna MC, Fiskum G (2016a) Sex-dependent mitochondrial respiratory impairment and oxidative stress in a rat model of neonatal hypoxic-ischemic encephalopathy. J Neurochem 137:714–729

    Article  CAS  Google Scholar 

  • Demarest TG, Waite EL, Kristian T, Puche AC, Waddell J, McKenna MC, Fiskum G (2016b) Sex dependent mitophagy and neuronal death following rat neonatal hypoxia-ischemia. Neuroscience 335:103–13

  • Finck BN, Kelly DP (2006) PGC-1 coactivators: inducible regulators of energy metabolism in health and disease. J Clin Invest 116:615–622

    Article  CAS  Google Scholar 

  • Hill CA, Fitch RH (2012) Sex differences in mechanisms and outcome of neonatal hypoxia-ischemia in rodent models: implications for sex-specific neuroprotection in clinical neonatal practice. Neurol Res Int 2012:867531

    Article  Google Scholar 

  • Hota KB, Hota SK, Chaurasia OP, Singh SB (2012) Acetyl-L-carnitine-mediated neuroprotection during hypoxia is attributed to ERK1/2-Nrf2-regulated mitochondrial biosynthesis. Hippocampus 22:723–736

    Article  CAS  Google Scholar 

  • Land JM, Booth RF, Berger R, Clark JB (1977) Development of mitochondrial energy metabolism in rat brain. Biochem J 164:339–348

    Article  CAS  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) Method. Methods 25:402–408

    Article  CAS  Google Scholar 

  • McKinney EA, Oliveira MT (2013) Replicating animal mitochondrial DNA. Genet Mol Biol 36:308–315

    Article  CAS  Google Scholar 

  • Mohagheghi F, Ahmadiani A, Rahmani B, Moradi F, Romond N, Khalaj L (2013a) Gemfibrozil pretreatment resulted in a sexually dimorphic outcome in the rat models of global cerebral ischemia-reperfusion via modulation of mitochondrial pro-survival and apoptotic cell death factors as well as MAPKs. J Mol Neurosci 50:379–393

    Article  CAS  Google Scholar 

  • Mohagheghi F, Khalaj L, Ahmadiani A, Rahmani B (2013b) Gemfibrozil pretreatment affecting antioxidant defense system and inflammatory, but not Nrf-2 signaling pathways resulted in female neuroprotection and male neurotoxicity in the rat models of global cerebral ischemia-reperfusion. Neurotox Res 23:225–237

    Article  CAS  Google Scholar 

  • Perez-Pinzon MA, Stetler RA, Fiskum G (2012) Novel mitochondrial targets for neuroprotection. J Cereb Blood Flow Metab 32:1362–1376

    Article  CAS  Google Scholar 

  • Pesce V, Fracasso F, Cassano P, Lezza AM, Cantatore P, Gadaleta MN (2010) Acetyl-L-carnitine supplementation to old rats partially reverts the age-related mitochondrial decay of soleus muscle by activating peroxisome proliferator-activated receptor gamma coactivator-1alpha-dependent mitochondrial biogenesis. Rejuvenation Res 13:148–151

    Article  CAS  Google Scholar 

  • Pesce V, Nicassio L, Fracasso F, Musicco C, Cantatore P, Gadaleta MN (2012) Acetyl-L-carnitine activates the peroxisome proliferator-activated receptor-gamma coactivators PGC-1alpha/PGC-1beta-dependent signaling cascade of mitochondrial biogenesis and decreases the oxidized peroxiredoxins content in old rat liver. Rejuvenation Res 15:136–139

    Article  CAS  Google Scholar 

  • Picca A, Lezza AM (2015) Regulation of mitochondrial biogenesis through TFAM-mitochondrial DNA interactions: useful insights from aging and calorie restriction studies. Mitochondrion 25:67–75

    Article  CAS  Google Scholar 

  • Rice JE III, Vannucci RC, Brierley JB (1981) The influence of immaturity on hypoxic-ischemic brain damage in the rat. Ann Neurol 9:131–141

    Article  Google Scholar 

  • Rosenthal RE, Williams R, Bogaert YE, Getson PR, Fiskum G (1992) Prevention of postischemic canine neurological injury through potentiation of brain energy metabolism by acetyl-L-carnitine. Stroke 23:1312–1317

    Article  CAS  Google Scholar 

  • Sato I, Konishi K, Mikami A, Sato T (2000) Developmental changes in enzyme activities and in morphology of rat cortex mitochondria. Okajimas Folia Anat Jpn 76:353–361

    Article  CAS  Google Scholar 

  • Scafidi S, Fiskum G, Lindauer SL, Bamford P, Shi D, Hopkins I, McKenna MC (2010a) Metabolism of acetyl-L-carnitine for energy and neurotransmitter synthesis in the immature rat brain. J Neurochem 114:820–831

    Article  CAS  Google Scholar 

  • Scafidi S, Racz J, Hazelton J, McKenna MC, Fiskum G (2010b) Neuroprotection by acetyl-L-carnitine after traumatic injury to the immature rat brain. Dev Neurosci 32:480–487

    CAS  Google Scholar 

  • Scarpulla RC (1997) Nuclear control of respiratory chain expression in mammalian cells. J Bioenerg Biomembr 29:109–119

    Article  CAS  Google Scholar 

  • Scarpulla RC (2002) Nuclear activators and coactivators in mammalian mitochondrial biogenesis. Biochim Biophys Acta 1576:1–14

    Article  CAS  Google Scholar 

  • Shadel GS, Clayton DA (1993) Mitochondrial transcription initiation. Variation and conservation. J Biol Chem 268:16083–16086

    CAS  Google Scholar 

  • Sharma J, Johnston MV, Hossain MA (2014) Sex differences in mitochondrial biogenesis determine neuronal death and survival in response to oxygen glucose deprivation and reoxygenation. BMC Neurosci 15:9

    Article  Google Scholar 

  • Smith AL, Alexander M, Rosenkrantz TS, Sadek ML, Fitch RH (2014) Sex differences in behavioral outcome following neonatal hypoxia ischemia: Insights from a clinical meta-analysis and a rodent model of induced hypoxic ischemic brain injury. Exp Neurol 254:54–67

    Article  Google Scholar 

  • Spinazzi M, Casarin A, Pertegato V, Salviati L, Angelini C (2012) Assessment of mitochondrial respiratory chain enzymatic activities on tissues and cultured cells. Nat Protoc 7:1235–1246

    Article  CAS  Google Scholar 

  • Suliman HB, Piantadosi CA (2016) Mitochondrial quality control as a therapeutic target. Pharmacol Rev 68:20–48

    Article  CAS  Google Scholar 

  • Virbasius JV, Virbasius CA, Scarpulla RC (1993) Identity of GABP with NRF-2, a multisubunit activator of cytochrome oxidase expression, reveals a cellular role for an ETS domain activator of viral promoters. Genes Dev 7:380–392

    Article  CAS  Google Scholar 

  • Waddell J, Hanscom M, Shalon Edwards N, McKenna MC, McCarthy MM (2016) Sex differences in cell genesis, hippocampal volume and behavioral outcomes in a rat model of neonatal HI. Exp Neurol 275:285–295.

  • Xie Y, Li J, Fan G, Qi S, Li B (2014) Reperfusion promotes mitochondrial biogenesis following focal cerebral ischemia in rats. PLoS One 9:e92443

    Article  Google Scholar 

  • Yin W, Signore AP, Iwai M, Cao G, Gao Y, Chen J (2008) Rapidly increased neuronal mitochondrial biogenesis after hypoxic-ischemic brain injury. Stroke 39:3057–3063

    Article  Google Scholar 

  • Zhang C, Wong-Riley MT (2000) Depolarizing stimulation upregulates GA-binding protein in neurons: a transcription factor involved in the bigenomic expression of cytochrome oxidase subunits. Eur J Neurosci 12:1013–1023

    Article  CAS  Google Scholar 

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Acknowledgments

All authors were directly involved in the design or conduct of the study and contributed to the writing/editing of the manuscript. TGD was responsible for data collection, analysis, interpretation, and manuscript/figure preparation. RAS and ELW assisted in data collection. JW performed all surgeries and drug treatments. GF and MCM were responsible for experimental design and interpretation of results.

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Authors and Affiliations

  1. Department of Anesthesiology and the Center for Shock, Trauma, and Anesthesiology Research (S.T.A.R.), University of Maryland School of Medicine, 685 West Baltimore Street, Bldg. MSTF 5-34, Baltimore, MD, 21201, USA

    T. G. Demarest, R. A. Schuh, E. L. Waite & Gary Fiskum

  2. Department of Pediatrics, University of Maryland School of Medicine, Baltimore, MD, 21201, USA

    J. Waddell & M. C. McKenna

  3. Program in Neuroscience, University of Maryland School of Medicine, Baltimore, MD, 21201, USA

    T. G. Demarest

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  1. T. G. Demarest
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  2. R. A. Schuh
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  3. E. L. Waite
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  4. J. Waddell
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  5. M. C. McKenna
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  6. Gary Fiskum
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Correspondence to Gary Fiskum.

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Funding

This work was supported by National Institutes of Health Grant 5P01 HD016596 and the University of Maryland School of Medicine M. Jane Matjasko research endowment.

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The authors declare no conflict of interest.

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Demarest, T.G., Schuh, R.A., Waite, E.L. et al. Sex dependent alterations in mitochondrial electron transport chain proteins following neonatal rat cerebral hypoxic-ischemia. J Bioenerg Biomembr 48, 591–598 (2016). https://doi.org/10.1007/s10863-016-9678-4

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  • DOI: https://doi.org/10.1007/s10863-016-9678-4

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