Richard Morrison, PhD is a Professor of Neurological Surgery and Biological Structure. His research focuses on molecular biology of neuronal cell death, proteomics and glial tumorgenesis.
He is the PI on a Neuroproteomics Institutional Center Core Grant from NINDS along with multiple other NIH grants.
- Characterization of neuronal cell death
- Characterization of mitochondrial dynamics and function in neurons
Overexpression of the neuron-specific Drp1 isoform that includes exon 3 (ISO1) increases mitochondrial respiratory capacity in neurons
Camelia A. Danilov, Takuma Uo, Daciana Margineantu, David Hockenbery, Richard S. Morrison
1) Department of Neurological Surgery, University of Washington, Seattle, WA
2) Fred Hutchinson Cancer Research Center, Seattle, WA
Our previous in vitro studies indicate that the mitochondrial fission protein Drp1 is required for neuronal survival. We also demonstrated that cultured cortical neurons express several alternatively spliced Drp1 transcripts with or without exon 3. Inclusion of exon 3 is specific to postmitotic neurons, while exon 3 exclusion was always observed in non-neuronal cells including astrocytes, fibroblasts and adult spinal cord progenitor cells. The importance of exon 3 to Drp1 function and hence mitochondrial function has not been investigated. In this study we hypothesized that overexpression of the neuron-specific Drp1 isoform that includes exon 3 (ISO1) increases mitochondrial respiratory capacity in mouse postnatal cortical neurons when compared with the non-neuronal Drp1 isoform (ISO3).
Basal oxygen consumption (OCR, nmoles/min/# cells ± SEM) and extracellular acidification rates (ECAR, mpH/min/#cells ± SEM) in neurons transfected with different Drp1 isoforms were measured using an XF24 Analyzer (Seahorse Bioscience). Neurons expressing ISO1 displayed the highest basal respiratory rate (8.3 ± 2.2% ) compared with ISO 3 (4.7 ± 1.5% ) and neurons infected with lentivirus expressing IRES-driven GFP only (CTR-GFP, 3.9 ± 0.6% , P< 0.19). Cellular respiration rate after addition of the mitochondrial uncoupler, CCCP (0.5 µM) showed a significant increase in neurons overexpressing ISO1 (173.00 ± 25.3% ) compared with ISO3 (110.0 ± 13.8% ) or CTR-GFP (87.75.0 ± 12.6% , P< 0.05).
Moreover, suppressing Drp1 activity by expressing a dominant negative form of Drp1 (Drp1K38A) decreased both the basal (2.7 ± 0.4% ) rate and the maximum respiration rate induced by addition of CCCP (78.00 ± 13.9% ). We did not observe any difference among groups in the basal rate of extracellular acidification (glycolysis). However, the extracellular acidification rate was increased after CCCP addition in neurons overexpressing ISO1 (188.3 ± 16.03% ) when compared with SO3 (116.2 ± 10.5% ) and CTR-GFP (105.07 ± 15.03% , P<0.05).
In marked contrast to differentiated neurons, ISO1 expression did not increase the basal or CCCP treated respiration rate in 3T3 cells. Our findings suggest that Drp1 ISO1 exerts unique neuron-specific functions that may be required to meet the high energy demands of neurons.
Supported by NIH grants RO1NS35533, RO1NS056031 and the lentivirus core facility supported by P30 NS055088
Class I histone deacetylase (HDAC) inhibitors preserve glial cell function and viability to protect white matter from ischemic injury
Selva Baltan, Sean P. Murphy, Camelia A. Danilov, Amelia Bachleda, and Richard S. Morrison
1) Department of Neurology, University of Washington
2) Department of Neurological Surgery, University of Washington
The importance of white matter (WM) injury to stroke pathology has been underestimated and this may have contributed to the failure to translate potential therapeutics into the stroke clinic. HDAC inhibitors, drugs for which there are abundant clinical trial data, are both neuroprotective and also promote ‘neuronal’ differentiation of progenitor cells, properties which make them ideal candidates for stroke therapy. In the isolated mouse optic nerve (MON) deprived of oxygen and glucose (OGD), we found that pan- and Class I specific HDAC inhibitors preserved axonal viability and also improved functional recovery when normal levels are restored. Neuroprotection correlatesd with preservation of axonal mitochondria and ATP levels in MON, and also with up-regulation of a glutamate transporter (GLT-1) in WM astrocytes. Manipulation of HDAC activity may be a viable strategy for stroke treatment.
- How does injury (stroke) and disease (Alzheimer’s) precipitate nerve cell damage, dysfunction and death?
- How is mitochondrial structure and function (energy production) regulated in neurons and glia?
- Are the molecular pathways that regulate mitochondrial structure and function compromised in response to brain injury and disease?
- Demonstrating a role for the p53 tumor suppressor protein in neuronal cell death.
- Identifying a key role for the apoptotic protein, Bax, in p53-mediated cell death signaling.
- Demonstrating a requirement for histone deacetylase activity in p53-mediated cell death signaling.
- Providing an initial characterization of the proteins regulating mitochondrial fission and fusion in neurons and glia.
- Demonstrating that neurons express unique, alternatively spliced forms of the proteins that regulate mitochondrial dynamics which confer the proteins with distinct activities.
- Primary Neuronal and Glial Cultures
- Various knockout mice are used to evaluate the role of apoptotic mediators
- Lentivirus production, purification and transduction
- Traditional biochemical techniques
Immunostaining and Imaging
- Polyacrylamide gel electrophoresis
- Western blot
- Protein, mRNA and DNA purification
- Caspase cleavage assays
Bioenergetics-oxygen consumption and glycolysis
- Fluorescent and bright field imaging on fixed and live cells and tissues
Bibliography of Selected Publications (selected from a total of 93 publications)
- Morrison R.S., Kinoshita Y., Johnson M.D., Guo W. and Garden G.A. p53-dependent cell death signaling in neurons. Neurochem Res, 28:15-27, 2003.
- Fu L., Abu-Khalil A., Morrison R.S., Geschwind D.H. and Kornblum H. I. Expression patterns of epidermal growth factor receptor and fibroblast growth factor receptor 1 mRNA in fetal human brain. J Comp Neurol., 462:265-273, 2003.
- Yamaguchi F., Morrison R.S., Gonatas N.K., Takahashi H., Sugisaki Y., Teramoto A. Identification of MG-160, a FGF binding medial Golgi sialoglycoprotein, in brain tumors: An index of malignancy in astrocytomas. Int J Oncol., 22:1045-1049, 2003.
- Yu L-R., Issaq H.J., Conrads T.P., Uo T., Blonder J., Janini G.M., Morrison R.S. and Veenstra T.D. Evaluation of Liquid Chromatography-Mass Spectrometry for Routine Proteome Analyses. Journal of Liquid Chromatography & Related Technologies, 26(20):3325-3336, 2003.
- Yu L-R., Conrads T.P., Uo T., Issaq H.J., Morrison R.S. and Veenstra T.D. Evaluation of the acid-cleavable isotope-coded affinity tag reagents: application to camptothecin-treated cortical neurons. Journal of Proteome Research, 3:469-477 2004.
- Johnson M.D., Li-Rong Y., Conrads T.P., Kinoshita Y., Uo T., Lee S.W., Smith D., Veenstra T., and Morrison R.S. Proteome Analysis of DNA Damage-Induced Neuronal Death Using High Throughput Mass Spectrometry. J. Biol. Chem., 279:26685-26697, 2004.
- Garden G.A., Guo W., Tun C., Jayadev S., Balcaitis S., Moeller T., and Morrison R.S.. HIV Associated Neurodegeneration Requires p53 in Neurons and Microglia. FASEB J., 18(10):1141-1143, 2004. For Full Text: http://www.fasebj.org/cgi/doi/10.1096/fj.04-1676fje
- Yu LR, Conrads T.P., Uo T., Kinoshita Y., Morrison R.S., Lucas D.A., Chan K., Blonder J., Issaq H.J., and Veenstra T.D. Global Analysis of the Cortical Neuron Proteome. Mol. and Cell. Proteomics, 3:896-907, 2004.
- Garden G.A. and Morrison R.S. The Multiple Roles of p53 in the Pathogenesis of HIV Associated Dementia. BBRC, 331:799-809, 2005.
- Uo. T., Kinoshita Y., and Morrison R.S. Neurons exclusively express an alternatively spliced BH3 domain only Bak isoform, N-Bak, that promotes neuronal apoptosis. J. Biol. Chem., 280:9065-9073, 2005.
- Veenstra T.D., Conrads T.P., Hood B.L., Avellino A.M., Ellenbogen R.G., and Morrison R.S. Biomarkers: Mining the Biofluid Proteome. Molecular and Cellular Proteomics. 4:409-418, 2005.
- Johnson M.D., Yu L-R, Conrads T.P., Kinoshita Y., Uo T., McBee J.K., Veenstra T.D., and Morrison R.S. The Proteomics of Neurodegeneration. American Journal of PharmacoGenomics., 5:259-70, 2005.
- Elias MC, Tozer KR, Silber JR, Mikheeva S, Deng M, Morrison RS, Manning TC, Silbergeld DL, Glackin CA, Reh TA, Rostomilly RC: TWIST is expressed in human gliomas and promotes invasion. Neoplasia, 7:824-37, 2005.
- La Spada A.R. and Morrison R.S. The Power of the Dark Side: Huntington’s Disease Protein and p53 Form a Deadly Alliance. Neuron, 47: 1-3, 2005.
- Sikorski E.M., Uo T., Morrison R.S., and Agarwal A. Pescadillo interacts with the cadmium response element of the Human heme oxygenase-1 promoter in renal epithelial cells. J. Biol. Chem., 281:24423-30, 2006.
- Kinoshita Y. Uo T., Jayadev S., Garden G.A., Conrads T.P., Veenstra T.D. and Morrison, R.S. Potential Applications and Limitations of Proteomics in the Study of Neurological Disease. Archives of Neurology, 63:1692-1696, 2006.
- Tun C., Guo W., Nguyen H., Yun B., Libby R.T., Morrison R.S., Garden G.A. Activation of the extrinsic caspase pathway in cultured cortical neurons requires p53-mediated down-regulation of the X-linked inhibitor of apoptosis protein to induce apoptosis. J. Neurochem., 102:1206-1219, 2007.
- McBee J.K., Yu L-R, Kinoshita Y, Uo T., Beyer R.P., Veenstra T.D., and Morrison R.S. Proteomic Analysis of Protein Expression Changes in a Model of Gliomagenesis. Proteomics Clin. Appl., 1:1485–1498, 2007.
- Uo T., Kinoshita Y. and Morrison R.S. Apoptotic actions of p53 require transcriptional activation of PUMA and do not involve a direct mitochondrial/cytoplasmic site of action in postnatal cortical neurons. J. Neurosci, 27:12198-12210, 2007.
- Jayadev S., Yun B., Nguyen H., Yokoo H., Morrison R.S. and Gwenn A. Garden. The Glial Response to CNS HIV Infection Includes p53 Activation and Increased Expression of p53 Target Genes. J. Neuroimmune Pharmacology, 2:359-30, 2007.
- Morrison, R.S. and Veenstra, T.D. Biomarker discovery: Has it
been worth it so far? Proteomics – Clin. Appl., 2:1375-1378,
- Young JE, Taylor, JP, Garden GA, Martinez R, Tanaka F, C. Sandoval M, Smith AC, Sopher BL, Lin A, Fischbeck KH, Ellerby LM, Morrison RS, and La Spada AR. Polyglutamine-expanded androgen receptor truncation fragments activate a Bax-dependent apoptotic cascade mediated by JNK, c-Jun, and DP5/Hrk. J. Neurosci, 29:1987-1997, 2009.
- Uo T., Veenstra T.D., Morrison R.S. Histone Deacetylase Inhibitors Prevent p53-dependent and Independent Bax-Mediated Neuronal Apoptosis Through Two Distinct Mechanisms. J. Neurosci, 29:2824-2832, 2009.
- Uo, T., Dworzak J., Kinoshita, C., Kinoshita, Y., Inman, D.M., Horner, P.J. and Morrison, R.S. Drp1 levels constitutively regulate mitochondrial dynamics and viability in cortical neurons. (Cover illustration). Exp. Neurology, 218:274-285, 2009.
- Mihkeev A., Stoll E, Mikheeva S, Maxwell JP, Ray S, Morrison R S., Uo T. , Horner P, Rostomily R. A syngeneic glioma model to assess the impact of neural progenitor target cell age on tumor malignancy. Aging Cell, 8: 499-501, 2009.
- Baltan S., Inman D.M., Danilov C., Morrison R.S., Calkins D.J. and Horner P.J. Metabolic vulnerability disposes retinal ganglion cell axons to dysfunction in a model of glaucomatous degeneration. (Cover illustration). J. Neurosci, 30: 5644-5652, 2010.
- Waybright T., Avellino A.M., Ellenbogen R.G., Hollinger B.J., Yi M., Stephens R.S., Morrison R.S., Veenstra, T.D. Proteomic Characterization of Human Ventricular Cerebrospinal Fluid From Patients with Hydrocephalus. J. Proteomics, 73:1156-1162, 2010.
- Baltan S., Danilov CA., Bachleda A., Murphy S.P. and Morrison R.S. Class I Histone Deacetylase Inhibitors Preserve Glial Cell Function and Viability to Protect White Matter From Ischemic Injury. Submitted, 2010.
- Jayadev S., Case A., Eastman A.J., Nguyen H. Pollak J. Wiley J.C., Moller T., Morrison R.S. and Garden G.A. Presenilin 2 is the Predominant gamma-secretase in Microglia and Modulates Cytokine Release. PLoS ONE, 5:e15743, 2010.
- Jayadev S, Nesser NK, Hopkins S, Myers SJ, Case A, Lee RJ, Seaburg LA, Uo T, Murphy SP, Morrison RS, Garden GA. Transcription factor p53 influences microglial activation phenotype. Glia. 2011. PMID:21598312 .
- Baltan S., Danilov CA., Bachleda A., Murphy S.P. and Morrison R.S. Class I Histone Deacetylase Inhibitors Preserve Glial Cell Function and Viability to Protect White Matter From Ischemic Injury. J. Neurosci, 31(11): 3990-3999, 2011
- Declines in Drp1 and Parkin Expression Underlie DNA Damage-Induced Changes in Mitochondrial Length and Neuronal Death. J. Neurosci, 33(4): 1357-1365, 2013
Lab Contact Information
For more information about the Morrison Lab, call (206) 543-9763.
Published Research Articles
View complete lists of current research publications by faculty from the Department of Neurological Surgery.