Parkinson's and Alzheimer's Diseases Group

Introduction | Achievements | Objectives | Techniques | Collaborators | Publications | Contacts

Culvenor Neurodegeneration Laboratory located in the Centre for Neuroscience in collaboration with the Department of Pathology.

Staff / Students

Janetta Culvenor

Laboratory Head Dr Janetta G. Culvenor Research Fellow Dr Yuh Ping Chong PhD students Sonia George Ryan Mills Joe Sim

Introduction

Research involves study of the expression and localization of key proteins associated with neurodegeneration particularly the amyloid precursor protein, presenilins, and synucleins, as well as their interacting partners. Techniques involve development and characterization of antibodies for detection by biochemical analysis and immunocytochemical analysis. Studies include analysis by Western blotting of cultured cell extracts, animal tissue, and human brain. Laser confocal microscopy and immunofluorescence is used for study of intracellular localization of proteins in cultured cells. Immunogold labelling and transmission electron microscopy is utilized for fine structural analysis.

Achievements

• Localization of amyloid precursor protein after over-expression in cultured cells and yeast by immunoelectron microscopy
• Localization of presenilin to the endoplasmic reticulum-Golgi intermediate compartment of neuronal cells using laser confocal microscopy and immunolabelling in combination with metabolic inhibitors
• Establishment of presenilin 1 knock-out mouse stem cells to examine role of presenilin 1
• Generation of alpha-synuclein antibodies and detection of aggregates and cleavage products in cultured cells and human brain tissue

Objectives

• Study of nature of and localization of presenilin protein complexes and identification of novel components
• Study of role of nicastrin in brain and muscle protein complexes
• Investigation of truncated a-synuclein species isolated from cultured cells and from human brain extracts to identify forms of a-synuclein which may be causing toxicity
• Establishment of cell culture model of intracellular a-synuclein aggregation to examine the process of aggregation and modifiers
• Study of animal models of Parkinson's disease

Techniques

Generation of antibodies, immunocytochemistry, electrophoresis, Western blotting, tissue culture of primary neurons and cell lines, animal dissection, and human brain, Laser confocal microscopy and immunofluorescence, Immunogold labelling and transmission electron microscopy. 

Collaborators

Local

• G. Evin, (Dept. of Pathology) C. McLean (Australian Neuroscience Facility) study of presenilin complexes and binding partners
• Q.X. Li, R. Cappai, K. Barnham (Dept. of Pathology) study of a-synuclein aggregation and metabolites
• Associate Professor Heung-Chin Cheng, Dr Nicholas Williamson and Dr Kipros Gabriel (Dept. Biochemistry and Molecular Biology, Bio21 Research Institute)

External

• Associate Professor Paul Baird, Centre for Eye Research Australia

Funding Sources

• ANZ Charitable Trusts
• Australian Brain Foundation
• Bethlehem Griffiths Research Foundation
• Michael J. Fox Foundation for Parkinson's Research
• Neurosciences Victoria Ltd
• NHMRC
• Prana Biotechnology Ltd

Recent Publications

George, S., Van den Buuse, M., Mok, S.S., Masters, C.L., Li, Q.-X., and CULVENOR, J. G. (2008) Alpha-Synuclein transgenic mice exhibit reduced anxiety-like behaviour. Exp. Neurol. Accepted 20-12-07 

Silveyra, M.-X., Evin, G., Montenegro, M.-F., Vidal, C.J., Martínez, S., CULVENOR, J.G., and Sáez-Valero, J. (2008) Presenilin-1 interacts with acetylcholinesterase and alters its enzymatic activity and glycosylation. Mol. Cell Biol. Accepted 14-2-08

Mills, R.D., Sim, C.H., Mulhern, T.D., CULVENOR, J.G. and Cheng, H.-C. (2008) Biochemical aspects of the neuprotective mechanism of PTEN-induced kinase-1 (PINK1)  J. Neurochem. Accepted 17-1-08

Li, Q.-X., Mok, S.S., Laughton, K.M., McLean, C.A., Cappai, R., Masters, C.L., CULVENOR, J.G., and Horne, M.K. (2007) Plasma a-synuclein is decreased in subjects with Parkinson’s disease. Exp. Neurol. 204, 583-577 

Melrose, H.L., Kent, C.B., Taylor, J.P., Dachsel, J.C., Hinkle, K.M., Lincoln, S.J., Mok, S.S., CULVENOR, J.G., Masters, C.L., Tyndall, G.M., Bass, D.I., Ahmed, Z., Andorfer, C.A., Ross, O.A., Wszolek, Z.K., DelleDonne, A., Dickson, D.W., and Farrer, M.J. (2007) A comparative analysis of Lrrk2 expression in mouse brain and Lewy Body disease. Neurosci. 147, 1047-1058

Fodero-Tavoletti, M.T., Smith, D.P., McLean, C.A., Adlard, P.A., Barnham, K.J., Foster, L.E., Leone, L., Perez, K., Cortes, M., CULVENOR, J.G., Li,Q.X., Laughton, K.M., Rowe, C.C., Masters, C.L., Cappai, R., and Villemagne, V.L. (2007) In vitro characterization of Pittsburgh compound-B binding to Lewy bodies. J. Neurosci. 27, 10365-10371

Sim, C.H., Lio, D.S.S., Mok, S.S., Masters, C.L., Hill, A.F., CULVENOR, J.G., and H.-C. Cheng (2006) C-Terminal truncation and Parkinson’s disease-associated mutations down-regulate the protein serine/threonine activity of PTEN-induced Kinase-1 Hum. Mol. Genet., 15, 3251-3262
 
CULVENOR, J.G., Ilaya, N.T., Ryan, M.T., Canterford, L., Hoke, D., Williamson, N.A., McLean, C.A., Masters, C.L., and Evin, G. (2004) Characterization of Presenilin Complex from Mouse and Human Brain using Blue Native Gel Electrophoresis Reveals High Expression in Embryonic Brain and Minimal Change in Complex Mobility with Presenilin Mutations. Eur. J. Biochem.271, 375-385.

Smith, M. J., Sharples, R.A., Evin, G., McLean, C.A., Dean, B., Pavey, G., Fantino, E., Cotton, R.G.H., Imaizumi, K., Masters, C.L., Cappai, R., and Culvenor, J.G. (2004) Expression of Truncated Presenilin 2 Splice Variant in Alzheimer's Disease, Bipolar Disorder, and Schizophrenia Brain Cortex. Molec. Brain Res. 127, 128-135.

Ilaya, NT., Evin, G., Masters, CL., and CULVENOR, JG. (2004) Nicastrin Expression in Mouse Peripheral Tissues is not Co-ordinated with Presenilin and is High in Muscle. J. Neurochem. 91, 230-237

Fodero, L.R., Mok, S.S., Losic, D., Martin, L.L., Aguilar, M-I., Barrow, C.J., Livett, B.G., and Small, D.S. (2004) a-7 Nicotinic acetylcholine receptors mediate an Ab 1-42-induced increase in the levels of acetylcholinesterase in primary cortical neurones. J. Neurochem. 88, 1186-1193.

CULVENOR, J.G., Rietze, R.L., Bartlett, P.F., Masters, C.L., and Li, Q.-X. (2002) Oligodendrocytes from Neural Stem Cells Express a-Synuclein: Increased Numbers from Presenilin 1 Deficient Mice. NeuroReport. 13, 1305-1308.

CULVENOR, J.G., Evin, G., Cooney, M.A., Wardan, H., Robyn A. Sharples, Maher, F., Reed, G., Diehlmann, A., Weidemann, A., Beyreuther, K., and Masters, C.L. (2000) Presenilin 2 expression in neuronal cells: Induction during differentiation of  embryonic carcinoma cells. Exp. Cell Res. 255,192-206.

CULVENOR, J.G., McLean, C.A., Cutt, S., Campbell, B.C.V., Maher, F., Jäkälä, P., Hartmann, T., Beyreuther, K., Masters, C.M., and Qiao-Xin Li. (1999) Non-Ab component of  Alzheimer’s disease amyloid (NAC) revisited: NAC and a-synuclein are not associated with Ab amyloid. Am. J. Path. 155, 1173-1181.

CULVENOR, J.G., Henry, A., Hartmann, T., Evin, G., Galatis, D., Friedhuber, A., Jayasena, U. L. H. R., Underwood, J.R., Beyreuther, K., Masters, C.L. and Cappai, R. (1998) Subcellular localization of the Alzheimer's disease amyloid precursor protein and derived peptides expressed in a recombinant yeast system. Amyloid Int. J. Exp. Clin. Invest. 5, 79-89.

CULVENOR, J.G., Maher, F., Evin, G., Malchiodi-Albedi, F., Cappai, R., Underwood, J.R., Davis, J.B., Karran, E.H., Roberts, G.W., Beyreuther, K. and Masters, C.L. (1997) Alzheimer's Disease-Associated Presenilin 1 in Neuronal Cells: Evidence for Localization to the Endoplasmic Reticulum-Golgi Intermediate Compartment. J. Neurosci. Res., 49, 719-731

Contacts

Janetta Culvenor
Phone: 8344 3990
Fax: 8344 4004
Email: janetta@unimelb.edu.au

Laboratory Photos