電気泳動的に分離したネズミ肝核蛋白の誘導能
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An archencephalic inductive effect of various mammalian tissues has been found in their constituent PNPs. The present experiment was performed to make clear the dominant component in PNP fraction which acts as an inductive agent by means of ionophoretic-separation method. The liver tissue of rat (Wistar King), Which has been known to be an archencephalic inductor when applied to act upon the presumptive ectoderm of Triturus gastrulae, was used as a donor material. The extirpated fresh liver was chilled with equal volume of 0.14M NaCl solution adjusted to pH 7.4. Then, the chilled tissue was homogenized with glass homogenizer after adding further two volumes of 0.14M NaCl solution. The extract was obtained by centrifugation for 30 minutes at 24, 000×g, and M/5 streptomycin-sulfate was added to the supernatant as to make M/50 in final concentration. After being kept for two hours, the resultant precipitate was separated from supernatant (non-PNP-fraction) by centrifugation for 20 minutes at 24, 000×g. For further purification, the precipitate was then washed with M/50 streptomycin-sulfate in 0.14M NaCl solution. Thus obtained fraction has been known to be highly pure PNP (Cohen, '55). Ionophoretic separation was made on Tiselius type electrophoretic apparatus. The typical electrophoretic pattern of the PNPs was obtained after 60 minutes with Na-diethylbarbiturate-HCl solution, of which ionic strength was 0.1 and pH 8.6, giving a current of 8 mA. The PNP fraction was electrophoretically foundto be composed of three components, whose mobilities were -7.7, -5.4 and -2.6cm^2/sec/volt×10^<-5>. The fast moving component (-7.7) and slow moving ones (-5.4 and -2.6) were separated each other by the use of automatic isolation syringe attached to the apparatus (Spinco Model-H) and fixed immediately with could 90% ethanol to test the inductive effects. The embryological test was made by the explantation method using the presumptive ectoderm isolated from mid-gastrulae of Triturus pyrrhogaster as a reactor system. Microscopical observation revealed that the fast moving component was a strong archencephalic inductor, and wholley lacked mesoderm-inductive effect. On the other hand, inductive power of the slow moving components was weaker, though it was characteristic for neural inductor. It is probable that the neural inductive agent of rat liver exists dominantly in-7.7 component of PNP fraction.
- 社団法人日本動物学会の論文
- 1961-10-15
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