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Nickolay Brustovetsky, PhD

Professor of Pharmacology and Toxicology

Research Interest 

My research is focused on mitochondrial bioenergetics, calcium signaling, and neuronal cell death in aging and age-related neurodegenerations.

Education

1992 – 1997

Postdoctoral Fellow | Institute for Physical Biochemistry | University of Munich Munich, Germany | Mentor: Professor Martin Klingenberg

1989 – 1992

Postdoctoral Fellow | Institute for Biophysics | Russian Academy of Science Mentor: Professor Yuri V. Evtodienko

1989

PhD, Biophysics | Institute for Physiology | USSR Academy of Science, Tashkent

1980 – 1989

Graduate Study in Biophysics | Institute for Biophysics |  USSR Academy of Science|  Pushchino, USSR

1975 – 1980

M.S. | Voronezh State University |  Voronezh, USSR

Honors

2015 – present

Showalter Scholar

2014 – present

Associate Editor, Frontiers In Chemistry, Cellular Biochemistry section 

2014 – present

Member, NIH Study Section NOMD

2014

Invited speaker, Annual Meeting Society for Neuroscience, Washington, DC 

2013 – present

Editorial Board Member, International Journal of Biochemistry and Molecular Biology

2012 – present

 Editorial Board Member, Journal of Drug Metabolism and Toxicology

2010 – 2011

Showalter Trust Fund Award

2001 – 2004

American Heart Association Scientist Development Grant

1998 – 1999

American Heart Association Postdoctoral Award

1980

Diploma with Distinction, Voronezh State University, Voronezh, USSR

Our lab studies the mechanisms of calcium deregulation, mitochondrial dysfunction, and neuronal loss in aging and age-related neurodegenerations.

We use transgenic and knock-in mouse models of Huntington’s disease and cyclophilin D-knockout mice    (Ppif -/-) to study the mechanisms of Huntington’s disease (HD) and the role of calcium deregulation and mitochondrial injury in HD progression. 

We utilize highly purified isolated brain (synaptic and non-synaptic) mitochondria to analyze the effect of aging on mitochondrial functions. To study isolated mitochondria, we use a computerized setup for simultaneous measurements of respiration, membrane potential, swelling, and calcium uptake by mitochondria. Using isolated mitochondria, we study the mechanisms of mitochondrial injury in aging, particularly, the role of the mitochondrial permeability transition pore. We also investigate the effect of aging on reactive oxygen species (ROS) production in mitochondria.

In addition to isolated mitochondria, we use cultured neurons derived from newborn and adult mice and rats. We use cellular respirometry (Seahorse technology), live-cell fluorescence wide-field and a laser spinning-disk confocal microscopy to investigate mitochondrial dysfunction, calcium deregulation, and ROS production in cultured neurons. We use electron microscopy to analyze mitochondrial morphological changes. We employ western blotting and immunocytochemistry to evaluate expression of different proteins and investigate the release of mitochondrial proteins involved in programmed cell death, apoptosis.

The overall goal of our lab is to determine the mechanisms of calcium deregulation, mitochondrial dysfunction, and neuronal death in aging and age-related neurodegenerations such as Huntington’s disease.

Figure 1.  Simultaneous measurements of respiration and membrane potential Δψm with isolated rat brain mitochondria.

Figure 2.  Recordings of mitochondrial membrane potential in individual isolated brain mitochondria loaded with a fluorescent dye Rhodamine 123.  A, fluorescence image of individual mitochondria attached to glass coverslip.  B, fluorescence traces showing normalized (F/F0) Rhodamine 123 signal indicative of mitochondrial polarization.

Figure 3. The 3D reconstruction of mitochondrial network in live cultured hippocampal neurons.

Figure 4.  The 3D reconstructions of individual mitochondrion in live hippocampal neuron exposed to glutamate.

Figure 5.  Oxygen consumption by cultured hippocampal neurons measured with Seahorse XF24 analyzer.

Figure 6.  Glutamate-induced calcium deregulation in neurons from 8-month-old FVB/NJ mice. 
a.
 immunostaining with anti-MAP2 antibody (green) to visualize neurons and staining with DAPI to show nuclei (blue). The large nuclei outside of neurons belong to glial cells. b-e, selected pseudocolor images of an adult mouse (8 month old) neuron (10 days in vitro) loaded with the Ca2+-sensitive dye Fura-2FF and treated with 10mM glutamate plus 10mM glycine. The images were taken at indicated time points. f, fluorescence traces from three individual adult neurons in the typical experiment are shown. Arrowheads on the red trace indicate when images shown in b-e were taken.

Selected Publications

  • Mantel, C.R., O’Leary, H. A., Chitteti, B.R., Huang, X., Cooper, S., Hangoc, G., Brustovetsky, N., Srour, E.F., Lee, M.R., Messina-Graham, S., Haas, D.M., Falah, N., Kapur, R., Pelus, L.M., Bardeesy, N., Fitamant, J., Ivan, M., Kim, K.-S., and Hal E. Broxmeyer, H.E. (2015) Enhancing hematopoietic stem cell transplantation efficacy by mitigating oxygen shock. Cell 161, 1553-1565. PMID: 26073944
  • Hamilton, J., Pellman, J.J., Brustovetsky, T., Harris, R., and Brustovetsky, N. (2015) Oxidative metabolism in YAC128 mouse model of Huntington’s disease. Hum. Mol. Gen. PMID: 26041817.
  • Pellman, J.J., Hamilton, J., Brustovetsky, T., and Brustovetsky, N. (2015) Ca2+ handling in isolated brain mitochondria and cultured neurons derived from the YAC128 mouse model of Huntington’s disease. J. Neurochem. 134, 652-667. PMID: 25963273 
  • Brustovetsky, N. (2015) Mutant huntingtin and elusive defects in oxidative metabolism and mitochondrial calcium Handling. Molecular Neurobiology doi: 10.1007/s12035-015-9188-0. PMID: 25941077
  • Lakhter, A.J., Hamilton, J., Dagher, P.C., Mukkamala, S., Hato, T., Dong, X.C., Lindsey D Mayo, L.D., Harris, R.A., Shekhar, A., Ivan, M., Brustovetsky, N., and Naidu, S.R. (2014) Ferroxitosis: a cell death from modulation of oxidative phosphorylation and PKM2-dependent glycolysis in melanoma. Oncotarget 5, 12694-12703. PMID: 25587028
  • Brustovetsky, T., Pellman, J.J., Yang, X.-F., Khanna, R., and Brustovetsky, N. (2014) Collapsin response mediator protein 2 interacts with NMDA receptor and Na+/Ca2+ exchanger and regulates their functional activity. J. Biol. Chem. 289, 7470-7482. PMID: 24474686
  • Ashpole, N.M., Chawla, A.R., Martin, M.P., Brustovetsky, T., Brustovetsky, N., and Hudmon, A. (2013) Loss of calcium/calmodulin-dependent protein kinase II activity in cortical astrocytes decreases glutamate uptake and induces neurotoxic release of ATP. J. Biol. Chem. 288, 14599-14611. PMID: 23543737
  • Brittain, M.K., Brustovetsky, T., Brittain, J.M., Khanna, R., Cummins, T.R., and Brustovetsky, N. (2012) Ifenprodil, a NR2B-selective antagonist of NMDA receptor, inhibits reverse Na+/Ca2+ exchanger in neurons. Neuropharmacology 63, 974-982. PMID: 22820271
  • Ashpole N.M., Song W., Brustovetsky T., Engleman E.A., Brustovetsky N., Cummins T.R., and Hudmon A. (2012) Calcium/calmodulin-dependent protein kinase II (CaMKII) inhibition induces neurotoxicity via dysregulation of glutamate/calcium signaling and hyperexcitability. J. Biol. Chem. 287, 8495-84506. PMID: 22253441
  • Brittain, M.K., Brustovetsky, T., Sheets, P. L., Brittain, J. M., Khanna, R., Cummins, T. R., and Brustovetsky, N. (2012) Delayed calcium dysregulation in neurons requires both the NMDA receptor and the reverse Na+/Ca2+ exchanger. Neurobiology of Disease, 46, 109-117. PMID: 22249110
  • Brittain, J.M, Chen, L., Wilson, S.M., Brustovetsky, T., Gao, X., Ashpole, N.M., Molosh, A.I., You, H., Hudmon, A., Shekhar, A., White, F.A., Zamponi, G.W., Brustovetsky, N., Chen, J., and Khanna, R. (2011) Neuroprotection against traumatic brain injury by a peptide derived from the collapsin response mediator protein 2 (CRMP-2). J. Biol. Chem. 286, 37778-37792. PMID: 21832084
  • Brittain, J.M, Duarte, D.B., Wilson, S.M., Zhu, W., Ballard, C., Johnson, P.L., Liu, N., Xiong, W., Ripsch, M.S., Wang, Y., Fehrenbacher, J.C., Fitz, S.D., Khanna, M., Park, C.-K., Schmutzler, B.S., Cheon, B.M., Due, M.R., Brustovetsky, T., Ashpole, N.M., Hudmon, A., Meroueh, S.O., Hingtgen, C.M., Brustovetsky, N., Ji, R.-R., Hurley, J.H., Jin, X., Shekhar, A., Xu, X.-M., Oxford, G.S., Vasko, M.R., White, F.A., and Khanna, R. (2011) Suppression of inflammatory and neuropathic pain by uncoupling CRMP-2 from the presynaptic Ca2+ channel complex. Nature Medicine, 17, 822-829. PMID: 21642979
  • Brustovetsky T, Brittain MK, Sheets PL, Cummins TR, Pinelis V, Brustovetsky N. KB-R7943, an inhibitor of the reverse Na+ /Ca2+ exchanger, blocks N-methyl-D-aspartate receptor and inhibits mitochondrial complex I. Br J Pharmacol. 2011 Jan;162(1):255-70. doi: 10.1111/j.1476-5381.2010.01054.x. PubMed PMID: 20883473; PubMed Central PMCID: PMC3012420.
  • Brustovetsky T, Li T, Yang Y, Zhang JT, Antonsson B, Brustovetsky N. BAX insertion, oligomerization, and outer membrane permeabilization in brain mitochondria: role of permeability transition and SH-redox regulation. Biochim Biophys Acta. 2010 Nov;1797(11):1795-806. Epub 2010 Jul 23. PubMed PMID:20655869; PubMed Central PMCID: PMC2933961.
  • Brustovetsky T, Bolshakov A, Brustovetsky N. Calpain activation and Na+/Ca2+ exchanger degradation occur downstream of calcium deregulation in hippocampal neurons exposed to excitotoxic glutamate. J Neurosci Res. 2010 May 1;88(6):1317-28. PubMed PMID: 19937813; PubMed Central PMCID: PMC2830319.
  • Storozhevykh TP, Senilova YE, Brustovetsky T, Pinelis VG, Brustovetsky N. Neuroprotective effect of KB-R7943 against glutamate excitotoxicity is related to mild mitochondrial depolarization. Neurochem Res. 2010 Feb;35(2):323-35. Epub 2009 Sep 22. PubMed PMID: 19771515.
  • Li T, Brustovetsky T, Antonsson B, Brustovetsky N. Dissimilar mechanisms of cytochrome c release induced by octyl glucoside-activated BAX and by BAX activated with truncated BID. Biochim Biophys Acta. 2010 Jan;1797(1):52-62. Epub 2009 Aug 5. PubMed PMID: 19664589; PubMed Central PMCID: PMC2788011.
  • Brustovetsky T, Li V, Brustovetsky N. Stimulation of glutamate receptors in cultured hippocampal neurons causes Ca2+-dependent mitochondrial contraction. Cell Calcium. 2009 Jul;46(1):18-29. Epub 2009 May 5. PubMed PMID: 19409612; PubMed Central PMCID: PMC2703686.
  • Li V, Brustovetsky T, Brustovetsky N. Role of cyclophilin D-dependent mitochondrial permeability transition in glutamate-induced calcium deregulation and excitotoxic neuronal death. Exp Neurol. 2009 Aug;218(2):171-82. Epub 2009 Feb 21. PubMed PMID: 19236863; PubMed Central PMCID: PMC2710407.
  • Li T, Brustovetsky T, Antonsson B, Brustovetsky N. Oligomeric BAX induces mitochondrial permeability transition and complete cytochrome c release without oxidative stress. Biochim Biophys Acta. 2008 Nov;1777(11):1409-21. Epub 2008 Aug 15. PubMed PMID: 18771651; PubMed Central PMCID: PMC2613194.
  • Bolshakov AP, Mikhailova MM, Szabadkai G, Pinelis VG, Brustovetsky N, Rizzuto R, Khodorov BI. Measurements of mitochondrial pH in cultured cortical neurons clarify contribution of mitochondrial pore to the mechanism of glutamate-induced delayed Ca2+ deregulation. Cell Calcium. 2008 Jun;43(6):602-14. Epub 2007 Nov 26. PubMed PMID: 18037484.

Department of Pharmacology and Toxicology | 635 Barnhill Drive, MS A401 | Indianapolis, IN 46202