Intracellular Distribution of Estradiol and Estrone in Rat Uterus
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Uteri from 3 rats were incubated in vitro with 3H estrone(E1)or 3H estradiol(E2)at 4°C or 37°C. After incubation, uteri were taken up from incubation medium a, t a certain time interval, washed and then homogenized. The homogenates were separated to cytosol and nuclear pellet by centrifugation. The nuclear pellet was extracted with 0.4M KCl. The extract was called as nuclear extract and the residue as pellet.The total uptake of radioactivity (cytosol+nuclear extract+pellet) was always higher at incubation with 3HE2 than with 3HEl.The percentage of radioactivity in each fraction was calculated. Concerning E2, 80% was in cytosol and 20% in nuclear fraction at 4°C incubation, while 60% was in cytosol and 40% in nuclear fraction at 37°C incubation. Our nuclear fraction did not contain microsome and mitochondria which might be a reason why our value of nuclear fraction was smaller than the value reported by Jensen et al.On the other hand, as fbr E1 90% was in cytosol and 10% in nuclear fraction at 4°C incubation, while 80% was in cytosol and 20% in nuclear fraction at 37°C incubation. From these results, translocation of E1 to nucleus is also temperature dependent as E2 but judging from the difference between at 4°C and at 37°C incubation, it's less temperature dependent as for E1 than E2. As compared with the value of E2, it is considered more difficult for estrone to translocate to nucleus than for E2.When free estrogen was eliminated by dextran charcoal treatment, over half of radioactivity recognized in cytosol and nuclear extract was diminished. This diminution rate was more abundant than the rate expected from the results of the previous experiments with sucrose density gradient analysis. More exact method was considered to he necessary to eliminate free estrogen, Considerable radioactivity was also recognized in the residue of nuclear fraction after extraction with 0.4M KCl. This radioactivity could not be decided as meaningless, since 2S estrone binding substance could bc extracted from this residue with 2% triton as reported previously.
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