Rift Valley 熱 Virus 感染のマウス肝劃分活性度に及ぼす影響 第1篇 マウス肝劃分法に対する吟味

In order to investigate the interaction between Rift Valley Fever virus and host mouse liver cells the author has used cell fractionating method as introduced by Hogeboom-Schneider. Since the results of cell fractionation vary depending on experimental procedures, it is important to examine various conditions such as concentration of sucrose, addition of calcium chloride, period for homogenizing, washing of fractionatants etc. In the present study, the author has tried to obtain an ideal fractionating method for experimental purposes and it,was noted that manometrical studies with Hogeboom-Schneider's method showed very constant results with high recovery values. Using succinate for substrate nuclear,mitochondrial and supernatant with microsome fractions showed 13 to 18%, 65 to 70% and 5 to 11% of the entire homogenate activity. Recovery values were 91 to 98%. After 3 to 4 repeated washings of nuclear fraction constant' succinoxidase activity was observed. With mitchondrial fraction repeated washings caused a gradual loss of succinoxidase activity. In electron microscopic examination 0.7 to 0.9 micron spherical mitochondria were observed using 0.25 M sucrose solution as fractionating media but 0.3 to 0.7x0.9 micron rod-shaped mitochondria were mixed with spherical particles when 0.88M sucrose solution was used. As compared with the morphological pects of mitochondria observed in the liver sections, mitochondria fractionated in 0.88M sucrose media were considered to be in a more physiological condition. It was also noted that the addition of calcium chloride to the fractionating media appeared to make tissue homogenizing and fractionating procedures easier