STUDY OF SOME FACTORS THAT AFFECT THE DISTRIBUTION OF NUCLEAR VOLUMES IN PREPARATIONS OF RAT LIVER NUCLEI

Abstract

The effect of the homogenization procedure, the centrifugation scheme, and the composition of the suspension medium on the distribution of nuclear volumes has been studied.It has been shown that the Waring Blendor not only destroys a greater number of the nuclei during homogenization, but also that this destruction is a selective one. At neutral pH values, no direct relationship appears to exist between the DNA content of the nuclei and their density. For this reason, purification in concentrated sucrose solutions produces a selective loss of the lighter nuclei, which includes small diploid stromal nuclei and some of the larger polyploid type of parenchymal nuclei.The study of the effect of increasing the calcium and magnesium ion concentrations (from 0.001 to 0.005 M) on the nuclear distribution showed that these ions produce a selective shrinkage and condensation of the nuclei, probably through different mechanisms.