Summary
Mevinolin, an inhibitor of cholesterol synthesis, was used to study the effect of endogenous cholesterol synthesis on the morphology and function of differentiating and differentiated fetal rat adrenocortical cells grown in primary culture. Upon adrenocorticotrophic hormone (ACTH) stimulation under conditions in which endogenous cholesterol synthesis was inhibited but exogenous (lipoprotein) cholesterol was available, the cells differentiated normally from glomerulosa-like to fasciculata-like cells; the steroid hormone secretion was maximally induced. Under conditions in which cholesterol synthesis was maximally inhibited by mevinolin and the cells had no access to exogenous cholesterol, the cells did not differentiate into fasciculata-like cells; the ACTH-induced steroid response was highly suppressed under these conditions. The addition of either human low-density lipoprotein (LDL) or high-density lipoprotein (HDL3) to the culture medium restored the ACTH-induced differentiation and steroid secretion. Thus, in the absence of exogenous cholesterol, endogenous cholesterol synthesis was a prerequisite for differentiation. In cultures grown in the presence of exogenous cholesterol and ACTH with mevinolin-inhibited cholesterol synthesis and high steroid output, an increase in cytoplasmic lipids was evident, suggesting upregulation of LDL and HDL receptors. The results also demonstrated that induction of phenotypic differentiation from glomerulosalike into fasciculata-like cells can proceed in the presence of a cholesterol synthesis inhibitor like mevinolin; this differentiation in the absence of endogenous cholesterol synthesis is accompanied by the appearance of cytoplasmic cholesterol ester droplets, typical of fasciculata cells.
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Alberts AW, Chen J, Kuron G, Hunt V, Huff J, Hoffman C, Rothrock J, Lopez M, Joshua H, Harris E, Patchett A, Monaghan R, Currie S, Stapley E, Albers-Schonberg G, Hensens O, Hirschfield J, Hoogsteen K, Liesch J, Springer J (1980) Mevinolin: a highly potent competitive inhibitor of hydroxymethylglutaryl-coenzyme A reductase and a cholesterol-lowering agent. Proc Natl Acad Sci USA 77:3957–3961
Bensch WR, Ingebritsen TS, Diller ER (1978) Lack of correlation between the rate of cholesterol biosynthesis and the activity of 3-hydroxy-3-methylglutaryl coenzyme A reductase in rats and in fibroblasts treated with ML-236B. Biochem Biophys Res Commun 82:247–254
Brown MS, Goldstein JL (1980) Multivalent feedback regulation of HMG CoA reductase, a control mechanism coordinating isoprenoid synthesis and cell growth. J Lipid Res 21:505–517
Brown MS, Goldstein JL (1985) The LDL receptor and HMG-CoA reductase — two membrane molecules that regulate cholesterol homeostasis. Curr Top Cell Regul 26:3–15
Brown MS, Goldstein JL (1986) A receptor-mediated pathway for cholesterol homeostasis. Science 232:34–47
Brown MS, Kovanen PT, Goldstein JL (1979) Receptor-mediated uptake of lipoprotein-cholesterol and its utilization for steroid synthesis in the adrenal cortex. Recent Prog Horm Res 35:215–257
Endo A, Tsujita Y, Kuroda M, Tanzawa K (1979) Effects of ML-236B on cholesterol metabolism in mice and rats: lack of hypocholesterolemic activity in normal animals. Biochim Biophys Acta 575:266–276
Fidge NH, Nestel PJ (1985) Identification of apolipoproteins involved in the interaction of human high density lipoprotein3 with receptors on cultured cells. J Biol Chem 260:3570–3575
Fidge N, Leonard-Kanevsky M, Nestel P (1984) The hormonal stimulation of high-density lipoprotein binding, internalization and degradation by cultured rat adrenal cortical celis. Biochim Biophys Acta 793:180–186
Fojo SS, Hoegg JM, Lackner KJ, Anchors JM, Bailey KR, Brewer HB Jr (1987) Adrenocortical function in type II hyperlipoproteinemic patients treated with lovastain (mevinolin). Horm Metabol Res 19:648–652
Havel RJ, Eder HA, Bragdon JH (1955) The distribution and chemical composition of ultracentrifugally-separated lipoproteins in human serum. J Clin Invest 34:1345–1353
Heikkilä P, Kahri AI, Ehnholm C, Kovanen PT (1989) The effect of low-and high-density lipoprotein cholesterol on steroid hormone production and ACTH-induced differentiation of rat adrenocortical cells in primary culture. Cell Tissue Res 256:487–494
Illingworth DR, Corbin D (1985) The influence of mevinolin on the adrenal cortical response to corticotropin in heterozygous familial hypercholesterolemia. Proc Natl Acad Sci USA 82:6291–6294
Kahri AI (1966) Histochemical and electron microscopic studies on the cells of the rat adrenal cortex in tissue culture. Acta Endocrinol 108:1–96
Kahri AI, Pesonen S, Saure A (1970) Ultrastructural differentiation and progesterone-14C metabolism in cultured cells of fetal rat adrenals under influence of ACTH. Steroidologia 1:25–64
Kahri AI, Heikkilä P, Ehnholm C, Ranki H, Kovanen PT (1989) Sequential expression of high and low density lipoprotein receptors in differentiating fetal rat adrenocortical cells in primary culture. Endocrinology 125:68–75
Kita T, Brown MS, Goldstein JL (1980) Feedback regulation of 3-hydroxy-3-methylglutaryl coenzyme A reductase in livers of mice treated with mevinolin, a competitive inhibitor of the reductase. J Clin Invest 66:1094–1100
Kovanen PT, Goldstein JL, Chappell DA, Brown MS (1980) Regulation of low density lipoprotein receptors by adrenocorticotropin in the adrenal gland of mice and rats in vivo. J Biol Chem 255:5591–5598
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Laue L, Hoeg JM, Barnes K, Loriaux DL, Chrousos GP (1987) The effect of mevinolin on steroidogenesis in patients with defects in the low density lipoprotein receptor pathway. J Clin Endocrinol Metab 64:531–535
Li AC, Tanaka RD, Callaway K, Fogelman AM, Edwards PA (1988) Localization of 3-hydroxy-3-methylglutaryl CoA reductase and 3-hydroxy-3-methylglutaryl CoA synthase in the rat liver and intestine is affected by cholestyramine and mevinolin. J Lipid Res 29:781–796
Mazzocchi G, Rebuffat P, Belloni AS, Gottardo G, Meneghelli V, Nussdorfer GG (1988) Effects of mevinolin, an inhibitor of cholesterol synthesis, on the morphological and functional responses of rat adrenal zona fasciculata to a prolonged treatment with 4-aminopyrazolo-pyrimidine. Anat Rec 221:700–706
O'Hare MJ, Neville AM (1973) The steroidogenic response of adult rat adrenocortical cells in monolayer culture. J Endocrinol 56:537–549
Oram JF, Brinton EA, Bierman EL (1983) Regulation of high density lipoprotein receptor activity in cultured human skin fibroblasts and human arterial smooth muscle cells. J Clin Invest 72:1611–1621
Orci L, Brown MS, Goldstein JL, Garcia-Segura LM, Anderson RGW (1984) Increase in membrane cholesterol: a possible trigger for degradation of HMG CoA reductase and crystalloid endoplasmic reticulum in UT-1 cells. Cell 36:835–845
Pathak RK, Luskey KL, Anderson RGW (1986) Biogenesis of the crystalloid endoplasmic reticulum in UT-1 cells: evidence that newly formed endoplasmic reticulum emerges from the nuclear envelope. J Cell Biol 102:2158–2168
Rebuffat P, Mazzocchi G, Nussdorfer GG (1987) Effect of longterm inhibition of hydroxy-methylglutaryl coenzyme A reductase by mevinolin on the zona fasciculata of rat adrenal cortex. A combined morphometric and biochemical study. Virch Arch [B] 54:67–72
Salmenperä M, Kahri AI (1976) Corticosterone, 18-OH-deoxycorticosterone, deoxycorticosterone and aldosterone secretion in tissue culture of foetal rat adrenals in the presence and the absence of ACTH. Acta Endocrinol 83:781–793
Verschoor-Klootwyk AH, Verschoor L, Azhar S, Reaven GM (1982) Role of exogenous cholesterol in regulation of adrenal steroidogenesis in the rat. J Biol Chem 257:7666–7671
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Heikkilä, P., Kahri, A.I., Kovanen, P.T. et al. Effects of mevinolin, an inhibitor of cholesterol synthesis, on the morphology and function of differentiating and differentiated rat adrenocortical cells in primary culture. Cell Tissue Res 261, 125–132 (1990). https://doi.org/10.1007/BF00329445
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DOI: https://doi.org/10.1007/BF00329445