2004 Fiscal Year Final Research Report Summary
Analysis of susceptibility to staphylococcal exfoliative toxin and gene expression of plakoglobin in a newborn and an eight-day-old mouse
| Project/Area Number |
14370798
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Laboratory medicine
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| Research Institution | JIKEI UNIVERSITY SCHOOL OF MEDICINE |
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Principal Investigator |
HOSHINA Sadayori Jikei Univeraity School of Med., Lab Med., Assistant Professor, 医学部, 助教授 (30119846)
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| Co-Investigator(Kenkyū-buntansha) |
KAMIDE Ryouichi Jikei Univeraity School of Med., Dermatology, Professor, 医学部, 教授 (40119780)
KOHNO Midori Jikei Univeraity School of Med., Lab Med., Assistant, 医学部, 助手 (00225385)
SAKURAI Susumu Kohno Clinical Medicine Research Institute, Chief Researcher, 室長 (20056542)
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| Project Period (FY) |
2002 – 2004
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| Keywords | Staphylococcal exfoliative toxin / ETA, ETB / Fibronectin / plakoglobin / β-catenin |
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| Research Abstract |
The gene coding the ETA substrate was cloned using the LA PCR in vitro cloning method, and the nucleotide sequence of the 280-bp fragment amplified by PCR was determined. This sequence showed a high degree of homology with the nucleotide sequence of the 3'-non-coding region of the β-catenin gene (128 of 133 nucleotids matched, 96%). Mouse β-catenin protein and plakoglobin, an intracellular plaque protein of desmosomes, are 56% identical. A comparison of β-catenin and plakoglobin gene expression using reverse transcriptase polymerase chain reaction(RT-PCR) in tissue from a newborn mouse and an eight-day old mouse revealed a significantly greater expression of β-catenin (264 bp in length) and plakoglobin (254 bp in length) in the newborn mouse, whereas expression of the β-catenin fragment was undetectable in the eight-day-old mouse. In addition, the expression of plakoglobin was either completely absent or present at very low levels in the eight-day-old mouse. Desmogleins, desmocollins, plectin, desmoyokin, and desmoplakin in Desmosomes have been detected at all stages of development in mice, including the embryonic stages, at birth, and 8 days after birth. Plakoglobin in the water-insoluble fraction prepared from ETA-injected newborn mouse skin tissues was present at low levels when compared (using immunoblotting) with that in a corresponding fraction prepared from control ETA-noninjected mouse skin tissues. However, the level of Dsg 1 in the water-insoluble fraction prepared from ETA-injected newborn mouse skin tissues was the same as that in the control ETA-noninjected newborn mouse skin tissues. Two enzyme-linked immunosorbent assays(ELISAs) were developed to detect Dsg 1 and plakoglobin using a double-antibody sandwich protocol. The ELISA procedures presented in this report may provide a sensitive method for the future detection of plakoglobin and Dsg 1 in clinical samples from patients with Staphylococcal scalded skin syndrome.
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