Control of steroid synthesis in human fetal adrenals in monolayer culture

Abstract

S. (1978) Control of steroid synthesis in human fetal adrenals in monol;myer culture. Can. 9. Bioclaern. 56,577-584The actions of adrenocorticotrophin 1-39 (ACTH), N6,02-dibutyryl adenosine-3',5'-cyclic phosphate (dbcAMP), and human chorionic gonadotrophin (hCG) on the sulfation of DHA and the synthesis of cortisol by human fetal adrenal cells in monolayer culture have been described. Electron microscopic studies indicate that the cells present in our cultures at the end of 5 days are intact and have many of the characteristics of secretory cells. Sulfation of DHA was strongly inhibited by 11-deoxycorticosterone, testosterone, androsterone, and dihydrotestosterone. Very small effects were observed with progesterone and corticosterone and insignificant effects with estrone, estradiol, and estriol. Androstenedione as well as 17a-hydroxylated corticosteroids showed no inhibition. When used as substrate, C,, A4-3-ketosteroids were good precursors of cortisol with 11-deoxycortisol being the best one followed by 17a-hydroxyprogesterone and progesterone. The conversion of progesterone (4pg12 ml) to cortisol was fivefold greater than under conditions where no substrate was present (endogenous production). ACTH in the concentration of 5 x Ulml added twice daily to the cell cultures for 8 days increased eightfold the sulfation of DHA and increased cortisol synthesis from progesterone 35-fold in 5 days. The addition of dbcAMP (5 x lW5 M ) or hCG used at 2.0 or 0.2 Ulml did not stimulate the sulfation of DHA. A lag period of at least 36 h is required before the stimulation of DHA sulfation or cortisol synthesis from progesterone can be observed. dbcAMP can stimulate the conversion of progesterone, 17a-hydroxyprogesterone, and I I-deoxycortisol to cortisol by the adrenal cells. These data suggest that 1 la-and 21-hydroxylation are stimulated by dbcAMP but 17a-hydroxylation is not altered.

Effects of melanotropic peptides on fetal adrenal gland.

“…The present data extend earlier reports on the capability of the human fetal adrenal gland to synthesize glucocorticoids (24,25) and the responsiveness of this process to ACTH (26,27). The fetal gland also produces numerous steroids lacking glucocorticoid activity (24,25 (4).…”

Control of Adrenal Androgen Secretion*

“…Another possible mechanism could be AA-induced alteration of binding of ACTH or other peptides to adrenocortical membrane receptors analogous to the decrease in binding of cosyntropin to purified bovine adrenocortical cell membranes induced by cortisol (184). If such short loop feedback control exists physiologically, it would probably be negative control, as suggested by the above in vitro studies (177)(178)(179)(180). In this model, normal adrenarche, for example, could be conceptualized as a physiological partial loss of short loop feedback.…”

“…The sulfation of DHA was also studied in human fetal adrenal gland, using a monolayer culture (180). Sulfokinase activity was inhibited by testosterone, dihydrotestosterone (DHT), and androsterone.…”

Ultrastructural Characteristics of Adult Rat Adrenocortical Cells Maintained in vitro With and Without ACTH

“…In recent years the technique of tissue culture has been extensively utilized for studying adrenocortical biochemical functions, and attempts have been made to correlate simultaneous cytomorphic and biochemical responses. Thus, tissue explants from fetal or immature adrenals of a variety of species (Kahri 1966, Milner 1971, Gyevai et al 1972, Goodyear et al 1976, Kahri et al 1976, Baird et al 1978), and adrenocortical tissue from the adult rat and man have been successfully cultured Nussdorfer 1972, Biswas et al 1967, Armato et al 1974, Nussdorfer et al 1977. Recently, isolated cells from adult adrenals have also been cultured by several workers ( O H a r e and Neville 1973, Price et al 1975, Suyama et al 1977, Hornsby andGill 1978).…”