EFFECT OF IONIC-STRENGTH ON THE INTERACTION OF ANDROGENS WITH SERTOLI-CELL NUCLEI

Only 35-50% of the label accumulated after incubation of cultured Sertoli cells with 3H-testosterone was readily extractable with 0.4 M KCl during a 1 h exposure. The degree of extractability was relatively constant over the pH range 7.0-8.5 but could be increased by prolonged (15 h) exposure. While 0.1 M KCl extracted a measurable amount of label, 0.4 M KCl was significantly more efficient. Furthermore, a higher proportion of the material extracted with 0.4 M KCl was associated with macromolecular species. After a 45 min exposure to 3H-testosterone, the nuclear fraction contained primarily labeled testosterone and its 5 α-reduced metabolites. The relative distribution of these metabolites between salt-resistant and readily extractable forms varied between experiments. In contrast, 3H-R1881 (17β-hydroxy-17-methylestra-4,9,11-trien-3-one) remained essentially intact int the nuclear fraction but also was only 35% extractable with 0.4 M KCl. In conclusion, although the quantitative aspects of salt extractability appear to depend to some extent upon the extraction conditions, it is apparent that the Sertoli cell nuclear fraction accumulates a significant amount of androgen in a form which is relatively resistant to removal with 0.4 M KCl. The biological significance of this phenomenon reamins to be established. © 1979.