| Budget Amount *help |
¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 2001: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2000: ¥900,000 (Direct Cost: ¥900,000)
|
|---|
| Research Abstract |
McLeod syndrome is a rare X-linked neuromuscular diseases caused by mutations of KX gene, which produces 37-kDa membrane transport protein. This syndrome manifests acanthocytosis, Hunchington disease-like symptoms characterized by chorea, myopathy such as elevated serum creatine kinase level and proximal muscle weakness and atrophy. However, why these neuromuscular symptoms appear is still unknown. In the present study, we carried out molecular study of our experienced family of McLeod syndrome, and studied the expression of Kx protein and XK mRNA in the muscle and the central nervous system as follows : 1) XK gene analysis : We extracted genomic DNA from the patient and his family members with informed consent. PCR of three exons of XK gene was performed, and directly sequenced. The result revealed the patient had a novel one base (C) insertion mutation at codon 151 of the XK gene and produced 5' frameshift. His mother showed heterozygous for this mutation, whereas his father and olde
… More
r sister was normal. 2) Production of anti-human Kx peptide antibody : We produced synthetic peptide corresponding to the human Kx amino acid seuqnces as follows : KNGLSEEIEKEVGQAEG. We immunized this peptide to a rabbit and IgG fraction was purified. 3) Expression of KX in human biopsied muscles : We performed immunohistochemistry of the human biopsied muscle specimen by using anti-Kx peptide antibody. In normal muscles, Kx was mainly immunostained in the sarcoplasmic reticulum, but not in the sarcolemma. In McLeod muscle, expression of the sarcoplasmic reticulum seemed to be reduced. 4) Expression of KX in the rat central nervous systems : We performed immunohistochemistry of the rat central nervous system by using this antibody. Kx was expressed ubiquitously in the rat neuron, especially in the nuclear and cytoplasmic lesion (endoplasmic reticulum) but not in the cell membrane. There were no staining for axons or nerve fibers. In RT-PCR analysis of the XK gene, XK mRNA was ubiquitously expressed in the rat brain. Less
|
