Adrenal nuclear matrix isolation. A morphologic and biochemical study.

Abstract

The nuclear matrix of adrenal cells was isolated by using the methods proposed by Commerford et al and Kaufmann et al for the liver nuclear matrix isolation. Both methods permitted, to the best of our knowledge for the first time, to prepare the nuclear matrix of a steroidogenic cell and therefore to study some regulatory mechanisms governing steroidogenesis. Commerford et al's method retains nuclear envelope and so produces a higher contamination; Kaufmann et al's method presents a higher purity since the nuclear envelope was removed by Triton X-100. No RNase digestion has been employed for the isolation of the residual nuclear matrices. Both methods however, permit the isolation of fractions with a good morphology, retaining a reticular nucleolus, interchromatinic granules, and a fibrogranular scaffold extending from the nucleolus to the nuclear lamina. The major peptides detected by 1-D SDS-PAGE were 123, 56, 46 and 41 kDa; with both methods protein profiles were similar. Identification of proteins by immunodetection reveals lamins A and C, 80 and 65 kDa respectively; no labeling was found for actin (45 kDa) and vimentin (57 kDa). In short, adrenal nuclear matrix was isolated, Kaufmann et al's method being the method of choice.