细胞功能测定

Huntington’s disease (HD) and Friedreich’s ataxia (FRDA) are associated with defects of respiratory-chain enzyme activities. In the respective disorders, these can be identified in tissue samples from postmortem brain and also during life from skeletal or cardiac muscle samples. The ...

A protein marked for degradation by the ubiquitin-proteasome pathway (UPP) is attached to multiple molecules of ubiquitin, a 76-amino-acid protein that targets the protein for rapid hydrolysis by 26S proteasome. Impaired function of UPP results in accumulation of misfolded and ubiqu ...

The histologic staining technique for central nervous system (CNS) tissue known as the Golgi technique was initially developed more than 125 yr ago. It was with this technique that, for the first time, whole nerve cells and their processes were simultaneously observed microscopically. Al ...

Transcriptional dysregulation has emerged as an important pathologic mechanism underlying the pathogenesis of Huntington’s disease (HD). The control of transcription depends on appropriate binding of transcription factor proteins to specific promoter regions of genes. C ...

One of the characteristic findings in human Huntington’s disease (HD) is the alteration of neurotransmitter receptors. To a remarkable degree, transgenic HD mouse models recapitulate neurotransmitter receptor alterations. Neurotransmitter receptors can be assessed at the ...

Once into the expanded disease-associated range, trinucleotide repeat alleles become dramatically unstable in the germline and in somatic cells. The molecular mechanism(s) that underlie this unique form of dynamic mutation are poorly understood. Numerous transgenic mouse mod ...

This chapter describes the potential use of viral-mediated gene transfer in the central nervous system as a new strategy in developing animal models of neurodegenerative diseases. To illustrate the approach, procedures for the production of lentiviral vectors encoding polyQ prot ...

This chapter describes how transgenic mice can be made with human genomic DNA fragments cloned from DM1 patients’ DNA and how the CTG repeat instability is assessed over generations and in different tissues. Construction of cosmid libraries is fully reported from the extraction of high-mo ...

This review intends to provide the different DNA methods for diagnosis of the repeat in the FMR1 gene. The two DNA methods to determine the CGG repeat size are Southern blot hybridization and the polymerase chain reaction (PCR), including bisulfite treatment.

Huntington’s disease (HD) is a fatal neurodegenerative disorder caused by a CAG repeat expansion in the first exon of the HD gene. It encodes a protein known as huntingtin, which aggregates in the nuclei of affected neurons. These aggregates are an obvious therapeutic target, thus an organotypic ...

Common features underlie the generation and function of neurons in multicellular animals. It is likely that conserved pathways and genes also are involved in neuronal degeneration and malfunction. To address the molecular mechanisms of complex human neurological disorders, many ...

The expression of expanded polyglutamine in mammalian cells causes the formation of aggregates. For elucidation of the biochemical properties of the aggregates, isolation and solubilization of the aggregates are required. This chapter provides useful protocols for the solubi ...

Expansion of a homomeric stretch of glutamine residues beyond a critical threshold can produce neurodegenerative disease. This observation led to the idea that abnormal polyglutamine stretches can alter protein structure in ways that contribute to disease. Because they are prone to ...

Antibodies can be extremely useful tools for the field of triplet repeat diseases. These reagents are important for localizing proteins in tissues, and within cells, they can be used in the isolation and characterization of the components of protein complexes, they can distinguish protei ...

Myotonic dystrophy (DM1) is a neuromuscular disorder caused by a CTGn expansion in the 3′-untranslated region (UTR) of myotonic dystrophy protein kinase (DMPK). SIX5 is a homeodomain gene located just downstream of the repeat, and myotonic dystrophy WD protein (DMWD) is located close upstr ...

Small-pool polymerase chain reaction (PCR) constitutes the PCR amplification of a trinucleotide repeat in multiple small pools of input DNA containing in the order of from 0.5 to 200 genome equivalents. Products are resolved by agarose gel electrophoresis and detected by Southern blot hy ...

To facilitate identification of disease genes containing an expanded trinucleotide repeat, a repeat expansion detection (RED) and gene cloning system was established. The RED method was developed to enable detection of expanded trinucleotide repeat sequences in any DNA sample fr ...

The unusual genetic features of trinucleotide repeat (TNR) diseases have stimulated a substantial body of research into the underlying molecular mechanisms of repeat instability. As one useful tool to study TNR instability, selectable genetic assays for expansions and contract ...

Expansions of triplet repeats are responsible for more than 15 hereditary neurological disorders in humans (1,2). Triplet repeats are fairly stable when the number of elementary units is under approx 30, but become polymorphic in length with a clear bias for expansions when this threshold is e ...

Since their discovery in 1991, triplet repeat mutations have been found to be the cause of genomic fragile sites, two of which are linked to mental retardation, myotonic dystrophy, and several late-onset neurodegenerative diseases. In all cases, these mutations exhibit gametic and/or som ...