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Novel ways for improved cell replacement strategies in Parkinson’s disease: The potential of Nogo-A neutralization.

English title Novel ways for improved cell replacement strategies in Parkinson’s disease: The potential of Nogo-A neutralization.
Applicant Widmer Hans Rudolf
Number 135565
Funding scheme Project funding
Research institution Universitätsklinik für Neurochirurgie Inselspital Bern
Institution of higher education University of Berne - BE
Main discipline Neurophysiology and Brain Research
Start/End 01.02.2012 - 31.01.2016
Approved amount 331'000.00
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Keywords (7)

Parkinson's disease; Nogo-A; neuronal transplantation; neuronal phenotype; neurotrophic factors; dopaminergic neurons; signal transduction

Lay Summary (English)

Lead
Lay summary

Background

Parkinson’s disease (PD) is a neurodegenerative disorder of unknown etiology. The disease is characterized by major clinical disturbances such as resting tremor, bradykinesia and rigidity. The symptoms are caused by dopamine depletion in the striatum resulting from a progressive loss of the nigrostriatal dopamine producing (DAergic) neurons. While medication therapy provides adequate alleviation of the symptoms for several years, the long term treatment is complicated by progressive disability and development of severe side effects. Metabolic compromise, excitotoxicity and oxidative stress are considered the main mechanisms that cause neuronal cell death that is both of apoptotic and necrotic nature. Notably, a marked atrophy of dendrites and spines of striatal medium spiny neurons is observed in advanced PD indicating a dysfunctional signal integration in the striatofugal pathway. A therapeutic concept for long-term treatment of PD is the replacement of the degenerated DAergic neurons by transplanting cells releasing dopamine. Recent findings from our and other laboratories suggest that neuroprotective molecules and neurotrophic factors support the survival of neural DAergic cells and the engraftment of the transplant in the host brain. The myelin associated protein Nogo-A is one of the most potent growth inhibitors of the adult CNS. Nogo-A is expressed on the surface of oligodendrocytes and has been described to limit neuronal regeneration and plasticity. Importantly to note, Nogo-A is also expressed in subpopulations of neurons of the adult and developing CNS. Several findings hint to the idea that Nogo-A has also functions in addition to prevention of axonal regeneration and plasticity. So accumulating body of data demonstrated that Nogo-A has also a neuronal differentiation inhibitor function. Given that Nogo-A is expressed in different populations of neurons in the adult CNS and modulates the cell differentiation, further studies are needed to better understand the function and the expression of this protein in health and disease.

Aim and Experimental Design

At present only few studies have investigated the expression of Nogo-A and NgR1 in the nigrostriatal system. Our preliminary data gathered by means of double immunofluorescence demonstrated that a subpopulation of DAergic cells also express Nogo-A. We do not know, however, on the extent of colocalization of DAergic neurons with Nogo-A and/or Nogo-A receptor (NgR1) and on the DAergic cell subtype. Accordingly, our research group will assess on the expression pattern of Nogo-A and NgR1 in the nigrostriatal system of the rat brain during development and in the PD disease state. Furthermore, the effects of Nogo-A neutralization in rat brain cultures and characterization of the interactions with neurotrophic factors are foreseen. Finally, the potential of Nogo-A neutralization of donor tissue prior to transplantation will be studied on the outcome in a rat model of Parkinson’s disease.

Significance

We anticipate that the results of these experiments will yield new insights regarding the fundamental pathophysiological features in PD and will also form a cornerstone for improved cell replacement strategies for patients suffering from Parkinson’s disease. 

 

Direct link to Lay Summary Last update: 21.02.2013

Responsible applicant and co-applicants

Employees

Publications

Publication
Conditioned medium from Endothelial Progenitor Cells promotes number of dopaminergic neurons and exerts neuroprotection in cultured ventral mesencephalic neuronal progenitor cells
Di Santo Stefano, Seiler Stefanie, Ducray Angélique D., Widmer Hans Rudolf (2019), Conditioned medium from Endothelial Progenitor Cells promotes number of dopaminergic neurons and exerts neuroprotection in cultured ventral mesencephalic neuronal progenitor cells, in Brain Research, 1720, 146330-146330.
Neurotrophic factor-based strategies to enhance survival and differentiation of neural progenitor cells toward the dopaminergic phenotype
Di Santo Stefano, Widmer HansR (2018), Neurotrophic factor-based strategies to enhance survival and differentiation of neural progenitor cells toward the dopaminergic phenotype, in Brain Circulation, 4(3), 139-139.
Nogo-receptor 1 antagonization in combination with neurotrophin-4/5 is not superior to single factor treatment in promoting survival and morphological complexity of cultured dopaminergic neurons
Seiler Stefanie, Di Santo Stefano, Sahli Sebastian, Andereggen Lukas, Widmer Hans Rudolf (2017), Nogo-receptor 1 antagonization in combination with neurotrophin-4/5 is not superior to single factor treatment in promoting survival and morphological complexity of cultured dopaminergic neurons, in Brain Research, 1668, 56-64.
Antagonization of the Nogo-Receptor 1 Enhances Dopaminergic Fiber Outgrowth of Transplants in a Rat Model of Parkinson’s Disease
Seiler Stefanie, Di Santo Stefano, Andereggen Lukas, Widmer Hans R. (2017), Antagonization of the Nogo-Receptor 1 Enhances Dopaminergic Fiber Outgrowth of Transplants in a Rat Model of Parkinson’s Disease, in Frontiers in Cellular Neuroscience, 11, 151.
Simultaneous transplantation of fetal ventral mesencephalic tissue and encapsulated genetically modified cells releasing GDNF in a hemi-parkinsonian rat model of Parkinson’s disease
Perez-Bouza Alberto, Di Santo Stefano, Seiler Stefanie, Meyer Morten, Andereggen Lukas, Huber Alexander, Guzman Raphael, Widmer Hans R. (2017), Simultaneous transplantation of fetal ventral mesencephalic tissue and encapsulated genetically modified cells releasing GDNF in a hemi-parkinsonian rat model of Parkinson’s disease, in Cell Transplantation, 1572.
Nogo-A neutralization improves graft function in a rat model of Parkinson’s disease
Seiler S Di Santo S and Widmer HR (2016), Nogo-A neutralization improves graft function in a rat model of Parkinson’s disease, in Front. Cell. Neurosci, 10:87, 1.
Non-canonical actions of Nogo-A and its receptors
Seiler Stefanie, Di Santo Stefano, Widmer Hans Rudolf (2016), Non-canonical actions of Nogo-A and its receptors, in Biochem Pharmacol, (100), 28-39.
Characterization of FA1/dlk1 expressing cells in the rat nigrostriatal system and effects of a 6-OHDA lesion.
Liechti R, Ducray AD, Jensen P, Di Santo S, Seiler S, Jensen CH, Meyer M, Widmer HR (2015), Characterization of FA1/dlk1 expressing cells in the rat nigrostriatal system and effects of a 6-OHDA lesion., in PLoS One., 10(2), e0116088.
Effects of forskolin on Trefoil factor 1 expression in cultured ventral mesencephalic dopaminergic neurons
Jensen P, Ducray AD, Widmer HR, Meyer M (2015), Effects of forskolin on Trefoil factor 1 expression in cultured ventral mesencephalic dopaminergic neurons, in Neuroscience, 317, 699.
Effects of forskolin on Trefoil factor 1 expression in cultured ventral mesencephalic dopaminergic neurons
Jensen Pia, Heimberg Michel, Ducray Angelique D., Widmer Hans R., Meyer Morten (2015), Effects of forskolin on Trefoil factor 1 expression in cultured ventral mesencephalic dopaminergic neurons, in Neuroscience , 25(3), 156-156.
Loss of Nogo-A expressing neurons in a rat model of Parkinson’s disease.
Schawkat K, Di Santo S, Seiler S, Widmer HR (2015), Loss of Nogo-A expressing neurons in a rat model of Parkinson’s disease., in Neuroscience , 59-72.
Nogo-A and its functions beyond axonal inhibition: The controversial role of Nogo-A in Parkinsons’s disease
Seiler Stefanie, Widmer Hans Rudolf (2015), Nogo-A and its functions beyond axonal inhibition: The controversial role of Nogo-A in Parkinsons’s disease, in Neural Regen Res , 10(8), 1223.
Antagonizing Nogo-receptor 1 promotes the number of cultured dopaminergic neurons and elongates their neurites
Seiler Stefanie, Pollini Dario, Di Santo Stefano, Widmer Hans R. (2014), Antagonizing Nogo-receptor 1 promotes the number of cultured dopaminergic neurons and elongates their neurites, in NEUROREPORT, 25(3), 160-160.
The Secretome of Endothelial Progenitor Cells Promotes Brain Endothelial Cell Activity through PI3-Kinase and MAP-Kinase
Di Santo Stefano, Seiler Stefanie, Fuchs Anna-Lena, Staudigl Jennifer, Widmer Hans Rudolf (2014), The Secretome of Endothelial Progenitor Cells Promotes Brain Endothelial Cell Activity through PI3-Kinase and MAP-Kinase, in PLOS ONE, 9(4), e95731.
Trefoil Factor 1 in the nigrostriatal system of 6-hydroxydopamine-lesioned rats
Jensen Pia, Heimberg Michel, Ducray Angelique D., Widmer Hans R., Meyer Morten (2014), Trefoil Factor 1 in the nigrostriatal system of 6-hydroxydopamine-lesioned rats, in NEUROREPORT, 25(3), 156-156.
Directed Differentiation and Functional Maturation of Cortical Interneurons from Human Embryonic Stem Cells
Maroof Asif M., Keros Sotirios, Tyson Jennifer A., Ying Shui-Wang, Ganat Yosif M., Merkle Florian T., Liu Becky, Goulburn Adam, Stanley Edouard G., Elefanty Andrew G., Widmer Hans Ruedi, Eggan Kevin, Goldstein Peter A., Anderson Stewart A., Studer Lorenz (2013), Directed Differentiation and Functional Maturation of Cortical Interneurons from Human Embryonic Stem Cells, in CELL STEM CELL, 12(5), 559-572.
Expression of Trefoil factor 1 in the developing and adult rat ventral mesencephalon.
Jensen P., Heimberg M., Ducray A. D., Widmer H. R., Meyer M. (2013), Expression of Trefoil factor 1 in the developing and adult rat ventral mesencephalon., Cognizant Communication Corporation, NY, USA.
Expression of Trefoil Factor 1 in the Nigrostriatal System of Hemiparkinsonian Rats Versus Unlesioned Controls
Jensen P., Heimberg M., Ducray A. D., Widmer H. R., Meyer M. (2012), Expression of Trefoil Factor 1 in the Nigrostriatal System of Hemiparkinsonian Rats Versus Unlesioned Controls, Cognizant Communication Corporation, NY, USA.
Phosphocreatine Interacts with Phospholipids, Affects Membrane Properties and Exerts Membrane-Protective Effects
Tokarska-Schlattner Malgorzata, Epand Raquel F., Meiler Flurina, Zandomeneghi Giorgia, Neumann Dietbert, Widmer Hans R., Meier Beat H., Epand Richard M., Saks Valdur, Wallimann Theo, Schlattner Uwe (2012), Phosphocreatine Interacts with Phospholipids, Affects Membrane Properties and Exerts Membrane-Protective Effects, in PLOS ONE, 7(8), e43178.

Collaboration

Group / person Country
Types of collaboration
Dr. Pascal Senn, ORL Clinic, University Hospital Bern Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
- Exchange of personnel
Prof. Dr. Morten Meyer, Anatomy and Neurobiology, Odense Denmark (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
- Exchange of personnel
Dr. med. E. Hewer, Pathology, Univ. of Bern Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
Prof. Dr. V. Enzmann; Ophthalmology, Univ. Hospital, Bern Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Research Infrastructure
Prof. Dr. Lorenz Studer, Sloan-Kettering Institute for Cancer Research, New York United States of America (North America)
- Publication
Prof. D. Surbek & Prof. M. Mueller, Gynecology, Univ. Hospital, Bern Switzerland (Europe)
- Research Infrastructure
Prof. Dr. Javier Fandino, Neurosurgery, KSA Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
Prof. Dr. Stephen Leib, Laboratory Neuroinfectiology, Institute for Infectious Diseases, University Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Research Infrastructure
Prof. Martin E.Schwab, Brain Research Institute, Univ. of Zurich Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
Dr. Anis Mir, Novartis Pharma AG, Basel Switzerland (Europe)
- Industry/business/other use-inspired collaboration

Scientific events

Active participation

Title Type of contribution Title of article or contribution Date Place Persons involved
SSN Annual Conference Poster Nogo-A neutralization improves engraftment of dopaminergic neurons in a rat model of Parkinson’s disease 23.01.2016 Lausanne, Switzerland Widmer Hans Rudolf; Seiler Stefanie;
NECTAR meeting Talk given at a conference “Neutralization of Nogo-A in the host brain improves graft function in a rat model of Parkinson’s disease” 09.12.2015 Lund, Sweden Widmer Hans Rudolf; Seiler Stefanie;
SCRM Annual Meeting: from bench to bedside’, November 6th, 2015. Talk given at a conference co-organizer of meeting 06.11.2015 Bern, Switzerland Widmer Hans Rudolf;
Annual Meeting of the Department of Clinical Research Poster Nogo-A neutralization improves graft function in a rat model of mild Parkinson’s disease. 04.11.2015 Bern, Switzerland Seiler Stefanie; Widmer Hans Rudolf;
ASNTR meeting Poster Di Santo S, Fuchs A-L, Seiler S and Widmer HR (2015). Endothelial progenitor cell derived conditioned medium promotes brain microvascular cell viability after ischemia insult in vitro 30.04.2015 Clearwater, United States of America Seiler Stefanie; Widmer Hans Rudolf;
SSPT Annual Conference Poster A combination of Nogo-receptor 1 antagonization and NT4/5 administration is superior than single factor treatment in promoting cultured dopaminergic neurons. 23.04.2015 Bern, Switzerland Seiler Stefanie; Widmer Hans Rudolf;
BENEFRI workshop Talk given at a conference In vitro and in vivo models to study Parkinson’s disease 04.02.2015 Bern, Switzerland Widmer Hans Rudolf;
Annual Meeting Clinical Neuroscience Poster Improved yield of cultured dopaminergic neurons by a combination of NT4/5 administration and Nogo-receptor 1 antagonization 22.01.2015 Bern, Switzerland Seiler Stefanie; Widmer Hans Rudolf;
NECTAR Meeting Talk given at a conference " The secretome of endothelial progenitor cells supports microvascular cells viability " 27.11.2014 Galway, Ireland Widmer Hans Rudolf;
NECTAR meeting Talk given at a conference "Assessments of new strategies to improve yield of dopaminergic neurons in vitro: Effects of Nogo-receptor 1 antagonization and NT4/5 administration". 27.11.2014 Galway, Ireland Widmer Hans Rudolf;
SCRM Annual Meeting: new horizons Talk given at a conference co-organizer of the Kick-off meeting 21.11.2014 Bern, Switzerland Widmer Hans Rudolf;
Annual Meeting of the Department of Clinical Research Poster Effects of Nogo-receptor 1 antagonization in combination with Creatine or GDNF administration on cultured dopaminergic neurons. 05.11.2014 Bern, Switzerland Seiler Stefanie; Widmer Hans Rudolf;
FENS Meeting Poster Antagonization of Nogo-receptor 1 in combination with NT4/5 administration leads to higher dopaminergic cell densities as compared to single factor treatment 05.07.2014 Milan, Italy Seiler Stefanie; Widmer Hans Rudolf;
ABCD Colloquium, Univ. Bern and Fribourg Talk given at a conference “Effects of Nogo-receptor 1 antagonization on dopaminergic neurons” 29.04.2014 Bern, Switzerland Seiler Stefanie;
ASNTR meeting Poster Analysis of the effects of conditioned medium from endothelial progenitors on brain microvascular cells 28.04.2014 Clearwater, United States of America Widmer Hans Rudolf; Seiler Stefanie;
GCB Symposium Poster Increased Dopaminergic Cell Number And Elongated Neu-rites Promoted By Nogo-receptor 1 Antagonization 29.01.2014 Bern, Switzerland Seiler Stefanie; Widmer Hans Rudolf;
Annual Meeting of the Swiss Society for Neuroscience Poster Effects of antagonizing Nogo-1 receptor on survival and morphological differentiation of dopaminergic neurons in mesencephalic cultures. 24.01.2014 Bern, Switzerland Widmer Hans Rudolf; Seiler Stefanie;
SSN Annual Conference Poster Effects of Nogo-receptor 1 antagonization in combination with neurotrophic factor administration on number of cultured dopaminergic neurons. 24.01.2014 Fribourg, Switzerland Seiler Stefanie; Widmer Hans Rudolf;
Annual Meeting of the Department of Clinical Research Poster Increased dopaminergic cell number and neurite length by antagonizing Nogo-1 receptor 06.11.2013 Bern, Switzerland Seiler Stefanie; Widmer Hans Rudolf;
INTR12 Talk given at a conference Antagonizing Nogo-receptor 1 promotes the number of cultured dopaminergic neurons and elongates their neurites 03.09.2013 Cardiff, Great Britain and Northern Ireland Seiler Stefanie; Widmer Hans Rudolf;
ASNTR meeting Poster Increased neurogenesis in the rat subventricular zone by infusion of soluble factors derived from endothelial progenitor cells. 25.04.2013 Clearwater, United States of America Widmer Hans Rudolf; Seiler Stefanie;
9th Annual Meeting Swiss Stem Cell Network Poster Promotion of angiogenesis of brain endothelial cells induced by conditioned medium treatmen critically involves the PI3-kinase pathway. 08.02.2013 Bern, Switzerland Seiler Stefanie; Widmer Hans Rudolf;
9th Annual Meeting Swiss Stem Cell Network Poster From blood to brain: Contribution of endogenous bone marrow-deriverd mesenchymal stem cells to the brain repair after stroke 08.02.2013 Bern, Switzerland Widmer Hans Rudolf;
Swiss Stem Cell Network (SSCN) meeting 2013. Talk given at a conference co-organizer 08.02.2013 Bern, Switzerland Widmer Hans Rudolf;
Annual Meeting Swiss Society for Neuroscience Poster Antagonizing Nogo-A receptor NEP1-40 promotes survival and /or differentiation of cultured midbrain cells 02.02.2013 Geneva, Switzerland Seiler Stefanie; Widmer Hans Rudolf;
Swiss Inner Ear Research Association Meeting Talk given at a conference “Creatine treatment for neuronal and inner ear derived progenitor cells”. 20.12.2012 Bern, Switzerland Widmer Hans Rudolf;
Annual Meeting Clinical Neuroscience Poster Increased yield of dopaminergic neurons by antagonizing Nogo-receptor 1. 04.12.2012 Bern, Switzerland Widmer Hans Rudolf; Seiler Stefanie;
Annual Meeting of the Department of Clinical Research Poster NEP1-40 increases the number of dopaminergic neurons and their axon length in dissociated cultures of fetal ventral mesencephalon 13.11.2012 Bern, Switzerland Seiler Stefanie; Widmer Hans Rudolf;
Kick-off meeting ‘Stem Cell Research & Regenerative Medicine’ Talk given at a conference co-organizer 01.11.2012 Bern, Switzerland Widmer Hans Rudolf;
74th Meeting of the SSAHE Poster Paracrine factors released from endothelial progenitor cells demonstrate neuroprotection in an in vitro model of Parkinson’s disease. 07.09.2012 Zurich, Switzerland Seiler Stefanie; Widmer Hans Rudolf;


Knowledge transfer events

Active participation

Title Type of contribution Date Place Persons involved
School visit in the frame work of the Swiss Life Sciences program. “Regeneration im zentralen Nervensystem: Performances, exhibitions (e.g. for education institutions) 04.11.2015 Biel, Switzerland Widmer Hans Rudolf;
School visit in the frame work of the Swiss Life Sciences program. “Regeneration im zentralen Nervensystem“. Performances, exhibitions (e.g. for education institutions) 23.09.2015 Bern, Switzerland Widmer Hans Rudolf;
school visit in the frame work of the Swiss Life Sciences program. “Regeneration im zentralen Nervensystem: Perspektiven für die Neurochirurgie?“ Performances, exhibitions (e.g. for education institutions) 28.08.2014 Frutigen, Switzerland Widmer Hans Rudolf;
“Regeneration im zentralen Nervensystem: Perspektiven für die Neurochirurgie?“ school visit in the frame work of the Swiss Life Sciences program Performances, exhibitions (e.g. for education institutions) 04.03.2013 Köniz, Switzerland Widmer Hans Rudolf;


Communication with the public

Communication Title Media Place Year
Talks/events/exhibitions «Hirnzelltransplantation beim Morbus Parkinson: Wo stehen wir heute?» German-speaking Switzerland 2014
Media relations: print media, online media Lähmung rückgängig gemacht: «Die Ärzte waren etwas zu euphorisch» International 2014
Talks/events/exhibitions “Are creatine and green tea good for retinal cells and brain endothelial cells?”. German-speaking Switzerland 2013
Talks/events/exhibitions “Regeneration im zentralen Nervensystem: Perspektiven für die Neurochirurgie?“ German-speaking Switzerland 2013
Talks/events/exhibitions Schulprogramm Swiss Life Sciences German-speaking Switzerland 2012

Awards

Title Year
Molly and Bernard Sanberg Memorial Award, ASNTR 2018
Benoit Pochon Preis für beste Doktorarbeit; DKF 2016
Best Poster; SSN Meeting Lausanne 2016
'Travel grant for young scientists', NECTAR Meeting Lund, Sweden 2015
Travel awrad; 9th FENS Forum of Neuroscience, Milan, Italy 2014
Travel award; International Meeting for Neural Therapy and Repair, Cardiff 2013 2013

Associated projects

Number Title Start Funding scheme
112529 Cell transplantation therapy in parkinson's disease: the potential of creatine treatment and characterization of the graft-host brain interactions 01.07.2006 Project funding
64975 Cell replacement strategies in neurodegenerative diseases: characterization and optimization of survival and function of donor tissue 01.01.2002 Project funding
145003 A new Zeiss LSM 710 laser scanning microscope for the DCR LCI Core Facility 01.12.2012 R'EQUIP
52947 Transplantation of human fetal nigral tissue: improving tissue quality by in vitro manipulation 01.09.1998 Project funding

Abstract

1. BACKGROUNDParkinson's disease (PD) is a disabling neurodegenerative disorder of unknown etiology characterized by a predominant and progressive loss of dopaminergic (DAergic) neurons of midbrain. While medication therapy provides adequate alleviation of the symptoms for several years, the long term treatment is complicated by progressive disability and development of severe side effects. A therapeutic concept for long-term treatment of PD is the replacement of the degenerated dopaminergic neurons by transplanting cells releasing dopamine. Clinical experiences have revealed proof of principle but also demonstrated significant limitations which ask for optimization of protocols. The major problems for cell replacement strategies in PD are concerning the appropriate source of donor tissue, the poor survival of transplanted cells and the suboptimal DAergic innervation of the host striatum.2. WORKING HYPOTHESISRecent findings from our and other laboratories suggest that neuroprotective molecules and neurotrophic factors support the survival of neural dopaminergic cells and the engraftment of the transplant in the host brain. Accumulating evidence indicate that Nogo-A plays an essential role in modulating the processes of axonal growth and innervation. Therefore, the combination of the beneficial effects of neurotrophic factors, neuroprotective molecules and Nogo-A inhibition might open new scenarios for PD therapies. In the present research project we will explore if the transplantation approach based on intracerebral application of anti-Nogo-A antibodies and treatment with creatine and/or neurotrophins significantly ameliorate the outcome of current cell transplantations protocols in PD. 3. SPECIFIC AIMSAim I.Assessment of the expression pattern of Nogo-A and NgR1 in the nigrostriatal system of the rat brain during development and in the PD disease state.Aim II.Assessment of the effects of Nogo-A neutralization in fetal rat VM cultures and characterization of the interactions with neurotrophic factors.Aim III.Assessment of the effects of Nogo-A neutralization on VM GFP transplant function in the 6-OHDA rat model of PD. 4. EXPERIMENTAL DESIGN AND/OR METHODS Distribution of Nogo-A and NgR1 in the nigrostriatal system of rat and human brain will be assessed using immunohistochemistry. By means of in vitro models of PD the effects of Nogo-A neutralization will be assessed using immunohistochemistry followed by analyzes of morphology. HPLC will be used to measure levels of dopamine in cultures. Identification of interactions with neurotrophic factors and creatine will include Western blot analyzes, signal transduction assessments and PCR. Using the 6-OHDA rat model of PD, we will explore Nogo-A neutralization on VM GFP transplants function of rats by means of behavioral assessment, immunohistochemistry and Western blot analysis. Phenotype identification of surviving neurons in grafts with or without factor pretreatment will be studied in the same setting using immunohistochemical methods in combination with measures of dopamine levels in the host brain. The possible alterations of host brain cell proliferation and neurogenesis will similarly be investigated in the 6-OHDA rat model of PD by means of BrdU injections and histological analyses.5. EXPECTED VALUE OF THE PROPOSED PROJECTThe proposed experiments are expected to contribute to the development of improved strategies and to increase understanding of events after transplantation in cell replacement approaches in PD. They concern a highly actual field of neurotransplantation research. Relevance to our studies is furthermore given that the used antibodies are currently in clinical evaluation. The impact of our approach likewise holds up for neural transplantation using alternative tissue sources and to other CNS lesions.
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