细胞技术

Vertical arrays are microarrays that have complex mixtures of nucleic acids as array elements, and that are hybridized with single sequence probes. Like dot blots, many different experiments can be spotted on a single vertical array, allowing single genes to be compared across many conditi ...

A high-throughput approach to prokaryotic differential display has been developed. A large number of reverse transcription polymerase chain reactions (RT-PCR) are performed on total RNA isolated from induced and control bacterial cultures. Each RT-PCR reaction uses a single olig ...

We describe here the GeneCalling� method for the discovery of differentially expressed genes, both known and novel, from any species and with useful sequence information to determine the potential function of novel genes captured. The method relies on transcript visualization coupl ...

Ordered differential display (ODD) is one of the approaches that uses systematic, rather than random, sampling of transcripts for display and thereby provides means to browse through essentially all the transcripts in the compared mRNA pools. It is specifically adapted for small amounts ...

Since its invention in 1992, differential display (DD) has become the most commonly used technique for identifying differentially expressed genes because of its many advantages over competing technologies such as DNA microarray, serial analysis of gene expression (SAGE), and subt ...

Differential display (DD) is one of the most commonly used approaches for identifying differentially expressed genes. However, there has been lack of an accurate guidance on how many DD polymerase chain reaction (PCR) primer combinations are needed to display most of the genes expressed in a e ...

Huntington’s disease is a hereditary neurodegenerative disorder, characterized by motor, psychiatric, and cognitive symptoms. The genetic defect responsible for the onset of the disease, expansion of CAG repeats in exon 1 of the gene that codes for huntingtin, has been unambiguously i ...

This chapter describes methods for the isolation of specific cell types that reveal how and where expansion can occur. For the hereditary component of expansion, the male germ cell has proved useful in distinguishing processes that can contribute to expansion, as described in our article (N ...

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 ...