Properties of isolated frog liver and kidney nuclei

In order for a nuclear preparation to be used for analytical purposes, the method of isolation and composition of the suspension medium must be carefully examined. Accordingly, satisfactory techniques for the isolation of frog liver and kidney nuclei were developed. The medium for frog liver nuclei consisted of: 55% glycerol, 0.001 M magnesium chloride, 0.033 M sodium β‐glycerophosphate and/or 0.002 M KH2PO4, K2HPO4 (pH 6.8), however, the addition of 0.15 M sucrose was essential for satisfactory isolation of kidney nuclei. Inclusion of sucrose (0.15 M) in the isolation medium promoted nucleolar swelling and a decrease in nuclear volume in liver cell nuclei. Nucleolar migration and extrusion were noted in solutions with high cationic content. The morphological appearance of isolated nuclei was found to be extremely sensitive to the ionic strength of the isolation medium, as was the isolation procedure in toto. Effects were considered to be the result of precipitation and swelling of nucleoprotein. Dissociation of nucleoprotein was considered to be associated with temperature change. The uptake of supra‐vital dyes aided in recognition of the morphological alterations and was also an indicator of nuclear viability. Trypsin readily altered the nuclear membrane and a rapid decrease in nuclear density occurred, but the nucleolus remained intact. The diverse response of liver and kidney nuclei as compared with the nucleated red blood cells (a contaminant) to treatment with trypsin was noted and its implications discussed.