ABSTRACT
UNLABELLED: BACKGROUND AND ÐBJECTIVE: Loss of conformation and function of sufficient number of proteins with high aggregation capacity plays an important role in the pathogenesis of many neurodegenerative disorders (NDD). Due to a recent discovery of new array of proteins with the capacity to form aggregates of nonamyloid type, new NDD models as well as a new level of understanding in vivo models which are already exist is needed. DNA/RN A binding proteins - FUS and TDP-43 play a crucial role in the pathogenesis of some forms of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia. The objective of the study was to develop a new ALS transgenic model. MATERIAL AND METHODS: In cell culture experiments, we studied mutant FUS proteins capable to form intracellular deposits morphologically similar to those observed in the autopsy material of ALS patients. RESULTS AND CONCLUSION: We created a transgenic mice line, in which a pathogenic form of human FUS protein was expressed in the nervous system. That led to the aggregation of FUS protein in spinal cord and motor neurons with the following degeneration and development of a phenotype, similar to the human ALS disease phenotype, in young grown-up animals. This neurodegenerative phenotype corresponds to a great number of clinical manifestations of human ALS and is an adequate transgenic model of the disease.
Subject(s)
Amyotrophic Lateral Sclerosis/genetics , Disease Models, Animal , Mice , RNA-Binding Protein FUS/genetics , Amyotrophic Lateral Sclerosis/metabolism , Animals , Humans , Mice, Transgenic , Mutation , RNA-Binding Protein FUS/metabolism , Real-Time Polymerase Chain Reaction , Spinal Cord/metabolismABSTRACT
In the present study we have used a transgenic mice overexpressing an amyloidogenic protein, gamma-synuclein, in the nervous system to address the effect of dimebon on proteinopathy progression. Neuroprotective effect of chronic dimebon administration in these mice at organismal level was confirmed by the increased lifespan. Using histological and biochemical approaches we have demonstrated that dimebon reduced the number of amyloid inclusions in spinal cord of transgenic animals and decreased the content of ubiquitinated proteins in detergent-insoluble fractions. These effects are likely to occur at the level of aggregated protein species, since transgene expression was not altered. Thus, pathological protein aggregation serves as one of dimebon targets in neurodegeneration.
Subject(s)
Amyloidosis/drug therapy , Indoles/pharmacology , Neuroprotective Agents/pharmacology , RNA, Messenger/genetics , Ubiquitinated Proteins/genetics , gamma-Synuclein/genetics , Administration, Oral , Amyloidosis/genetics , Amyloidosis/metabolism , Amyloidosis/pathology , Animals , Brain/drug effects , Brain/metabolism , Brain/pathology , Disease Models, Animal , Flocculation , Gene Expression , Longevity/drug effects , Mice , Mice, Inbred C57BL , Mice, Transgenic , Molecular Targeted Therapy , RNA, Messenger/metabolism , Solubility , Spinal Cord/drug effects , Spinal Cord/metabolism , Spinal Cord/pathology , Ubiquitinated Proteins/antagonists & inhibitors , Ubiquitinated Proteins/metabolism , Ubiquitination , gamma-Synuclein/metabolismABSTRACT
A number of neurodegenerative disorders have recently been coalesced into a group of proteinopathies because of the similarity of molecular mechanisms underlying their pathogenesis. A key step in the development of proteinopathies is a structural change that triggers aggregation of proteins, which are intrinsically prone to form aggregates due to their physical and chemical properties. Present review is devoted to the recent progress in the field of proteinopathies with specific focus on properties of aggregate-prone proteins, main stages of the development of molecular pathology and the role of cellular clearance systems in progression of neurodegeneration. Recent modifications in the nomenclature of neurodegenerative diseases will also be addressed.
Subject(s)
Amyloidogenic Proteins/chemistry , Neurodegenerative Diseases/metabolism , Proteostasis Deficiencies/metabolism , Amyloidogenic Proteins/metabolism , Autophagy , Humans , Neurodegenerative Diseases/classification , Neurodegenerative Diseases/physiopathology , Oxidative Stress , Proteasome Endopeptidase Complex/metabolism , Protein Conformation , Protein Folding , Proteostasis Deficiencies/classification , Proteostasis Deficiencies/physiopathology , Reactive Oxygen Species/metabolism , Terminology as Topic , Time Factors , Ubiquitin/metabolismABSTRACT
Aggregation of proteins liable to assembling into fibrils with subsequent formation of amyloid incorporations is an important component in the pathogenesis of many neurodegenerative diseases. Dimebon, a Russian drug, reduces the content of detergent-insoluble fibrillar forms of synuclein, the main protein component of pathological incorporations in neurons of transgenic mouse strain used in the study.
Subject(s)
Indoles/administration & dosage , Neurodegenerative Diseases/drug therapy , Neurons/drug effects , Neuroprotective Agents/administration & dosage , gamma-Synuclein/genetics , Amyloidogenic Proteins/genetics , Amyloidogenic Proteins/metabolism , Animals , Detergents/chemistry , Gene Expression/drug effects , Male , Mice , Mice, Transgenic , Neurodegenerative Diseases/genetics , Neurodegenerative Diseases/metabolism , Neurons/metabolism , Spinal Cord/drug effects , Spinal Cord/metabolism , Ubiquitin/metabolism , Ubiquitinated Proteins/genetics , Ubiquitinated Proteins/metabolism , gamma-Synuclein/metabolismABSTRACT
Chromocenter DNA fragments of polytene chromosomes of Drosophila orena ovarian nurse cells were cloned from a region-specific library (Dore 1) in a plasmid vector to yield 133 clones. A total of 76 clones were selected and sequenced. The total length of the sequenced fragments was 23940 bp. Analysis with several software packages revealed various repetitive sequences among the fragments of the Dore 1 library, including mobile genetic elements (25 fragments homologous to various LTR retrotransposons, five fragments homologous to LINEs, three fragments homologous to Helitrons, one fragment homologous to Polinton, and one fragment homologous to the mini-me non-LTR retrotransposon), four minisatellites, a satellite (SAR_DM), the (TATATG)n simple sequence repeat, and a low-complexity T-rich repeat. Sequences homologous to protein-coding genes were also found in the Dore 1 library. Various repetitive DNA sequences and gene homologs were identified as conserved sequences of pericentric heterochromatin of polytene chromosomes of ovarian nurse cells in nine species of the melanogaster species subgroup.
Subject(s)
DNA , Drosophila/genetics , Heterochromatin/genetics , Polytene Chromosomes/genetics , Repetitive Sequences, Nucleic Acid , Sequence Homology, Nucleic Acid , Animals , Cell Nucleus/genetics , Centromere/genetics , DNA/analysis , DNA/genetics , Female , Gene Library , Genetic Vectors , Molecular Sequence Data , Ovary/cytology , Sequence Analysis, DNAABSTRACT
Gamma(gamma)-synuclein is a member of synuclein family of cytoplasmic and predominantly neuronal proteins found only in vertebrates. Gamma-synuclein is abundant in axons and presynaptic terminals of neurons localized in brain regions involved in emotions, learning and memory. However, the role of gamma-synuclein in these brain functions was not previously assessed. We have demonstrated for the first time that the loss of gamma-synuclein results in a significant increase in the level of orientation response in novel environment and decrease in the level of state anxiety.
Subject(s)
Anxiety/psychology , Exploratory Behavior , gamma-Synuclein/physiology , Animals , Anxiety/genetics , Mice , Mice, Inbred C57BL , Mice, Knockout , gamma-Synuclein/geneticsSubject(s)
Aging/physiology , Maze Learning , Memory, Short-Term , alpha-Synuclein/metabolism , Animals , Mice , Mice, Inbred C57BL , Mice, Knockout , alpha-Synuclein/geneticsSubject(s)
Cognition/drug effects , Indoles/pharmacology , Neurodegenerative Diseases/drug therapy , Neurodegenerative Diseases/metabolism , Neuroprotective Agents/pharmacology , Proteins/metabolism , Amyloid/chemistry , Amyloid/metabolism , Animals , Drug Discovery , Gliosis/drug therapy , Gliosis/metabolism , Humans , Indoles/therapeutic use , Mice , Motor Activity/drug effects , Neurodegenerative Diseases/etiology , Neurodegenerative Diseases/physiopathology , Neuroprotective Agents/therapeutic use , Protein Multimerization/drug effects , Protein Stability , Protein Structure, Quaternary , Proteins/chemistry , Synucleins/chemistry , Synucleins/metabolismABSTRACT
The effects of inbreeding and low temperature on the pattern of homologous chromosome synapsis in ovarian nurse cell nuclei of Drosophila melanogaster strains were studied. Exposure to decreased temperature (16 degrees C) caused a noticeable increase in the rate of asynapses of homologous chromosomes, whereas this effect was insignificant for F30 inbreeding generation. Long-term inbreeding has a substantial effect on the relative positions of chromosomes in the nurse cell nuclei. This is visually evident only in the interstrain hybrids between highly inbred strains LA (F923) and HA (F923) or between either strain and laboratory strain Canton S or the flies from the natural population, where abnormalities in homologous chromosome synapsis are observed in virtually all nuclei.
Subject(s)
Cell Nucleus/metabolism , Chromosome Pairing , Chromosomes/metabolism , Inbreeding , Ovary/metabolism , Animals , Cell Nucleus/ultrastructure , Chromosomes/genetics , Chromosomes/ultrastructure , Cold Temperature , Drosophila melanogaster , Female , Ovary/ultrastructure , Species SpecificityABSTRACT
Evolutionary rearrangements of pericentromeric heterochromatin among Drosophila melanogaster subgroup species have been investigated. Region-specific DNA library from Drosophila orena ovarian nurse cell chromocenter has been obtained by microdissection of polytene chromosomes. The probe was localized on ovarian nurse cells chromosomes of D. melanogaster subgroup species using fluorescent in situ hybridization. Sequences homologous to the sequences of the DNA-probe were found in chromocenter and pericentromeric regions of D. orena polytene chromosomes; and in all pericentromeric regions of other species with several exceptions. So, there was no labeling on one of the arms of D. simulans chromosome 2 but such sequences were present on telomere of D. erecta chromosome 3 and in regions adjacent to the brightly DAPI-stained heterochromatic blocks of D. yakuba, D. santomea and D. teissieri chromosomes 2 and 3. At S6 stage (secondary reticulate nucleus), the labeled chromatin in D. orena could be found mostly within a restricted area and no such chromatin could be detected throughout the rest of the nucleus. On the contrary, the labeled DNA was spread diffusely in the nuclei of other species at this stage.