Nicholas T. Potter (editor) Methods in molecular biology, volume�217. Neurogenetics: methods and protocols

Abstract

This volume highlights many of the contemporary methodological approaches used for the characterisation of neurologically relevant gene mutations and their protein products. The 32 chapters in this book cover methodological aspects of mutation detection and screening; these are divided into six sections that cover quantitative polymerase chain reaction (PCR), trinucleotide repeat detection, sequence-based mutation detection, molecular detection of imprinted genes, fluorescence in situ hybridisation (FISH), in vitro protein expression systems and studies of protein expression and function. Many contributions provide clear descriptions of “how to” carry out a particular analysis. These include PCR-based methodologies, for example, dosage analysis in Duchenne muscular dystrophy and spinal muscular atrophy, detection of trinucleotide repeat expansions in Huntington’s disease, autosomal dominant cerebellar ataxias and fragile X syndrome, multiplex PCR in the analysis of Tay Sach’s disease, single-strand conformation polymorphism analysis in ion channel disorders and denaturing high performance liquid chromatography in Rett syndrome. The protein truncation test (PTT) is widely used in diagnostic laboratories to detect mutations at the protein level. In this volume, its use in the analysis of neurofibromatosis type 1 (NF1) and in tuberous sclerosis (TS1 and TS2) is clearly described, together with useful information on the relative merits and practicalities of DNA- and RNA-based PTT. Gel-based methodologies are described, namely Southern blot analysis in the analysis of Charcot Marie Tooth disease and pulse field gel analysis in fascioscapulohumeral muscular dystrophy. The contribution of molecular cytogenetics describes the use of FISH in the detection of genomic rearrangements in myelinopathies. Mitochondrial DNA diseases are extensively covered, with separate chapters on the mitochondrial encephalomyopathies and Leber’s hereditary optic neuropathy. Each of these contributions includes a useful clinical description, an account of the problems encountered in a diagnostic setting, clear laboratory protocols, advice on technical considerations and explanatory notes on interpretation of results. Other chapters are purely methodological, describing different techniques for the identification and isolation of disease-causing genes. RED (repeat expansion detection), RAPID (repeat analysis, pooled isolation and detection) and MALDI (matrix assisted laser desorption/ionisation mass spectrometry) are again clearly presented as laboratory protocols. These, although of interest in clinical diagnostic laboratories, may be more relevant to research groups. The last section of this volume is devoted to in vitro expression systems and the study of protein expression and function. Most applicable to diagnostic laboratories are the previously mentioned chapters on the use of the PTT, together with a description of immunological analysis of protein expression in neuromuscular disease. More fundamental studies of protein expression and function include the use of COS-7 cells with regard to myelin proteolipid protein and the characterisation of in vitro kinetics of polyglutamine aggregation. The remit of this book was to describe methodologies. The editor states that it is aimed at a wide group of readers including those carrying out molecular diagnosis, research workers and clinicians. Although it would be impossible for all parts of the book to be of direct and daily relevance to each group, many parts would be of interest to all.