- Email: [email protected]
ES107 Multiple system atrophy
Educational Seminars
86
Monday, 6 June 2005, 08.0009.15, Room 28
Monday, 6 June 2005, 08.00 09.15, Room 29
ES106 Genomics & proteomics (methodology)
ES107 Multiple system atrophy
Chair: Edna Gr~inblatt, W~irzburg, Germany Co-Chair: Jan Grimm, Palo Alto, CA, USA
Chair: Gregor Worming, Innsbruck, Austria Co-Chair: Christopher J. Mathias, London, United Kingdom Co-Chair: Albert Ludolph, Germany
Course description: The course will review different methods for tile identification and isolation of neurons as well as RNA-arnplification strategies suited for microarray analysis of small samples in Par!dnson's disease.
~ G e n e expression study techniques in post-mortem tissue and blood samples E. Ch-iinblatt*
*Wiirzburg, Germany In recent years there is an increasing interest in mRNA gone expression study ofneurodegenerative diseases, as well as of animal models. Classical genetics and neurochemistry did not manage to reveal the etiology of Par!dnson's disease and other neurodegenerative disorders. It is postulated that these diseases are controlled by multiple genes. Many of these genes themselves are signalling molecules, each of which controls tile expression of a subset of downstream genes. Thus, analysing differential gone expression, or RNA genetics, has become one of the most widely practiced strategies for studying the more complicated biological systems. Still, there are many factors to consider before starting such a gone-expression profiling project, tn this paper I would like to discuas tile importance of the mRNA source one wants to examine, whether it is post-mortem tissue or blood samples. There are many t'actors influencing tile quality as well as the gene expression profiling of the mRNA. In addition, the mRNA isolation methods may influence tile results. Additionally, gene expression techniques will be presented and compared to one another for their positive and negative advantages. This method implies to discover a much more complicated network of proteins involving in neurodegenerative diseases, which might then reveal their etiology.
•
Genomics & proteomics (methodology)
J. Grimm*
*Palo Alto, CA, USA The extraordinary cellular complexity of the mammalian central nervous systent presents a significant challenge to genomic studies of neurodegenerative diseases. For instance, the dopaminergic neurons which selectively degenerate in Parkinson's disease constitute less than 1 of 107 cells in the brain. The cellular heterogeneity is further exacerbated by differential progression of pathological changes between individual neurons. Lowabundance transcripts within rare types of neurons are often undetectable in RNA isolated from whole brain or gross-dissected brain homogenates. Moreover, significant changes in observed levels of gene expression can be introduced by small alterations in tissue composition rather than gone regulation in tile cell type of interest. To overcome these limitations, various strategies for gene profile analysis of homogeneous neuron populations or even single neurons have been devised. In this seminar, different methods for the identification and isolation ofnenrons as well as RNA-amplification strategies suited for microarray analysis of small samples will be discussed.
Monday, 6 June 2005, 08.00-09.15, Room 30
ES108 Basic aspects of tauopathiest synucleopathies Chair: Phil@ Kahle, M~inchen, Germany
Co-Chair: Eva Maria Mandelkow, Hamburg, Germany Course description: The course will review the role of tau in mechanisms of ncurodegenerative disease. The faculty will discuss the toxidty of tan. Point mutations and genomic multiplications of tile 0~-synudein (aSYN) gone cause autosomal-dominant hereditary PD. Moreover, intcaneuronal aSYN inclusions constitute the hallmark lesions of a nunlber of neurodegenerative diseases, including PD, dementia with Lewy bodies and multiple system atrophy. The formation of aSYN fibrils is a process involving several critical intermediate steps, which are influenced by interacter proteins and molecular chaperones. The detailed understanding of aSYN amyloidogenesis may lead to the development of anti-aggregative compounds for the treatment of PD and related disorders. At the cellular level, high elevation of aSYN protein levels causes the formation of inclusion bodies and apoptosis, as exemplified in primary oligo dendrocytes derived from transgenic (PLP)h[wt]aSYN mice and by lentiviral delivery of aSYN. We are currently testing anti-aggregation and anfi-apoptosis agents in these novel culture models of multiple systenl atrophy. Finally, aSYN fibrillization occurs in aging transgenic mice, and is restricted to predilected brain regions similar to human 0~-synucleinoapthies. Spinal cord, brainstem, and midbrain pathology coincides with a progressive loss of locomotor function, ultimately causing premature death of the animals. Cortical pathology, especially in the amygdala, appears to account for tile cognitive decline in (Thyl)-h[A30P]aSYN mice. Taken together these biochemical, genetic, and pathological insights have identified aSYN fibril formation as a pivotal cause of neurodegeneration and therefore, a formidable drug target.
~ T a u
and Tauopathy: Physiological role and pathological dysfunction
E.M. Mandelkow*
*Hamburg, Germany Tau is a microtubule-associated protein (MAP) located mostly in axons. Its main function is the control of stability and dynamics of microtubNes which form the tracks of axonal transport and are important for neurite outgrowth. In AD, tan occurs in a hyperphosphorylated and aggregated form. It can be phosphorylated by several ldnases whose role is a matter of debate. Most attention is fucussed on GSK-3b, cdk5, MAP ldnase (which phosphorylate Ser-Pro or Thr-Pro motifs) and M A R K (which phosphorylates tile KXGS motifs in tile microtubule-binding domain). Phosphorylation at Ser-Pro motifs tends to weaken the interaction of tan with microtubules, but the effect is often not very pronounced. On tile other hand, phosphorylation of the KXGS motifs by the kinase MARK/PAR1 detaches tau from microtubules and makes them labile. This type of phosphorylation occurs early in neurodegeneration. One enigmatic aspect of tan is its "natively unfolded" nature which is due to its hydrophilic arninoacid composition and makes it highly soluble. It is often assumed
86
Monday, 6 June 2005, 08.0009.15, Room 28
Monday, 6 June 2005, 08.00 09.15, Room 29
ES106 Genomics & proteomics (methodology)
ES107 Multiple system atrophy
Chair: Edna Gr~inblatt, W~irzburg, Germany Co-Chair: Jan Grimm, Palo Alto, CA, USA
Chair: Gregor Worming, Innsbruck, Austria Co-Chair: Christopher J. Mathias, London, United Kingdom Co-Chair: Albert Ludolph, Germany
Course description: The course will review different methods for tile identification and isolation of neurons as well as RNA-arnplification strategies suited for microarray analysis of small samples in Par!dnson's disease.
~ G e n e expression study techniques in post-mortem tissue and blood samples E. Ch-iinblatt*
*Wiirzburg, Germany In recent years there is an increasing interest in mRNA gone expression study ofneurodegenerative diseases, as well as of animal models. Classical genetics and neurochemistry did not manage to reveal the etiology of Par!dnson's disease and other neurodegenerative disorders. It is postulated that these diseases are controlled by multiple genes. Many of these genes themselves are signalling molecules, each of which controls tile expression of a subset of downstream genes. Thus, analysing differential gone expression, or RNA genetics, has become one of the most widely practiced strategies for studying the more complicated biological systems. Still, there are many factors to consider before starting such a gone-expression profiling project, tn this paper I would like to discuas tile importance of the mRNA source one wants to examine, whether it is post-mortem tissue or blood samples. There are many t'actors influencing tile quality as well as the gene expression profiling of the mRNA. In addition, the mRNA isolation methods may influence tile results. Additionally, gene expression techniques will be presented and compared to one another for their positive and negative advantages. This method implies to discover a much more complicated network of proteins involving in neurodegenerative diseases, which might then reveal their etiology.
•
Genomics & proteomics (methodology)
J. Grimm*
*Palo Alto, CA, USA The extraordinary cellular complexity of the mammalian central nervous systent presents a significant challenge to genomic studies of neurodegenerative diseases. For instance, the dopaminergic neurons which selectively degenerate in Parkinson's disease constitute less than 1 of 107 cells in the brain. The cellular heterogeneity is further exacerbated by differential progression of pathological changes between individual neurons. Lowabundance transcripts within rare types of neurons are often undetectable in RNA isolated from whole brain or gross-dissected brain homogenates. Moreover, significant changes in observed levels of gene expression can be introduced by small alterations in tissue composition rather than gone regulation in tile cell type of interest. To overcome these limitations, various strategies for gene profile analysis of homogeneous neuron populations or even single neurons have been devised. In this seminar, different methods for the identification and isolation ofnenrons as well as RNA-amplification strategies suited for microarray analysis of small samples will be discussed.
Monday, 6 June 2005, 08.00-09.15, Room 30
ES108 Basic aspects of tauopathiest synucleopathies Chair: Phil@ Kahle, M~inchen, Germany
Co-Chair: Eva Maria Mandelkow, Hamburg, Germany Course description: The course will review the role of tau in mechanisms of ncurodegenerative disease. The faculty will discuss the toxidty of tan. Point mutations and genomic multiplications of tile 0~-synudein (aSYN) gone cause autosomal-dominant hereditary PD. Moreover, intcaneuronal aSYN inclusions constitute the hallmark lesions of a nunlber of neurodegenerative diseases, including PD, dementia with Lewy bodies and multiple system atrophy. The formation of aSYN fibrils is a process involving several critical intermediate steps, which are influenced by interacter proteins and molecular chaperones. The detailed understanding of aSYN amyloidogenesis may lead to the development of anti-aggregative compounds for the treatment of PD and related disorders. At the cellular level, high elevation of aSYN protein levels causes the formation of inclusion bodies and apoptosis, as exemplified in primary oligo dendrocytes derived from transgenic (PLP)h[wt]aSYN mice and by lentiviral delivery of aSYN. We are currently testing anti-aggregation and anfi-apoptosis agents in these novel culture models of multiple systenl atrophy. Finally, aSYN fibrillization occurs in aging transgenic mice, and is restricted to predilected brain regions similar to human 0~-synucleinoapthies. Spinal cord, brainstem, and midbrain pathology coincides with a progressive loss of locomotor function, ultimately causing premature death of the animals. Cortical pathology, especially in the amygdala, appears to account for tile cognitive decline in (Thyl)-h[A30P]aSYN mice. Taken together these biochemical, genetic, and pathological insights have identified aSYN fibril formation as a pivotal cause of neurodegeneration and therefore, a formidable drug target.
~ T a u
and Tauopathy: Physiological role and pathological dysfunction
E.M. Mandelkow*
*Hamburg, Germany Tau is a microtubule-associated protein (MAP) located mostly in axons. Its main function is the control of stability and dynamics of microtubNes which form the tracks of axonal transport and are important for neurite outgrowth. In AD, tan occurs in a hyperphosphorylated and aggregated form. It can be phosphorylated by several ldnases whose role is a matter of debate. Most attention is fucussed on GSK-3b, cdk5, MAP ldnase (which phosphorylate Ser-Pro or Thr-Pro motifs) and M A R K (which phosphorylates tile KXGS motifs in tile microtubule-binding domain). Phosphorylation at Ser-Pro motifs tends to weaken the interaction of tan with microtubules, but the effect is often not very pronounced. On tile other hand, phosphorylation of the KXGS motifs by the kinase MARK/PAR1 detaches tau from microtubules and makes them labile. This type of phosphorylation occurs early in neurodegeneration. One enigmatic aspect of tan is its "natively unfolded" nature which is due to its hydrophilic arninoacid composition and makes it highly soluble. It is often assumed