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Biphasic effect of ACTH on growth of rat adrenocortical cells in primary culture

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The proliferation rate of differentiating fetal rat adrenocortical cells was studied in primary culture. In this system, stimulation with ACTH induces differentiation of zona glomerulosa-like cortical cells into zona fasciculata-like cells. Incorporation of bromodeoxyuridine (BrdU) was studied immunocytochemically by use of anti-BrdU antibody, and the proliferation rate was counted from the monolayer colonies of adrenocortical cells. After 21 days of cultivation in the absence of ACTH, the proliferation rate of zona glomerulosa-like cells was 10%. The rate slowly declined to 1% at the age of 100 days during continuous cultivation in the absence of ACTH. Stimulation with ACTH induced a strong inhibition in the proliferation rate (down to 2% during the first 24 h). Treatment with ACTH during the following 48 h led to an extremely intense proliferation of adrenocortical cells at a proliferation rate of 25%. Continuous treatment with ACTH up to 100 days led to a persistent growth of adrenocortical cells, and a proliferation rate over 2-fold higher than in control cells cultivated in the absence of ACTH. Thus, ACTH is the principal growth-promoting factor also in vitro, as has been found in in vivo studies. This growth effect is mediated by a biphasic course; at the beginning of differentiation the effect is inhibitory and is followed by a persistent stimulation of the growth of adrenocortical cells.

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References

  • Armato U, Nussdorfer GG (1972) Tissue culture of rat decapsulated adrenal glands. A methodological, ultrastructural and morphometric investigation. Z Zellforsch 135:245–273

    Google Scholar 

  • Armato U, Andreis PG, Draghi E (1977) Dose-related persistent proliferogenic action of corticotrophin 1–24 in normal adult human adrenocortical cells in primary tissue culture. J Endocrinol 72:97–98

    Google Scholar 

  • Di Blasio AM, Fujii DK, Yamamoto M, Martin MC, Jaffe RB (1990) Maintenance of cell proliferation and steroidogenesis in cultured human fetal adrenal cells chronically exposed to adrenocorticotropic hormone: Rationalization of in vitro and in vivo findings. Biol Reprod 42:683–691

    Google Scholar 

  • Diderholm H, Hellman B (1960) The cell renewal in the rat adrenal studied with tritiated thymidine. Acta Pathol Microbiol Scand 49:82–88

    Google Scholar 

  • Farese RV, Reddy WJ (1963) Observations on the interrelations between adrenal protein, RNA and DNA during prolonged ACTH administration. Biochim Biophys Acta 76:145–148

    Google Scholar 

  • Fong LG, Bonney E, Kosek JC, Cooper AD (1989) Immunohistochemical localization of low density lipoprotein receptors in adrenal gland, liver, and intestine. J Clin Invest 84:847–856

    Google Scholar 

  • Ford JK, Young RW (1963) Cell proliferation and displacement in the adrenal cortex of young rats injected with tritiated thymidine. Anat Rec 146:125–137

    Google Scholar 

  • Gonchoroff NJ, Katzmann JA, Currie RM, Evans EL, Houck DW, Kline BC, Greipp PR, Loken MR (1986) S-phase detection with an antibody to bromodeoxyuridine. Role of DNase pretreatment. J Immunol Methods 93:97–101

    Google Scholar 

  • Gratzner HG, Leif RC, Ingram DJ, Castro A (1975) The use of antibody specific for bromodeoxyuridine for the immunofluorescent determination of DNA replication in single cells and chromosomes. Exp Cell Res 95:88–94

    Google Scholar 

  • Hansson HA, Nilsson A, Isgaard J, Billig H, Isaksson O, Skottner A, Anderson IK, Rozell B (1988) Immunohistochemical localisation of insulin-like growth factor I in the adult rat. Histochemistry 89:403–410

    Google Scholar 

  • Heikkilä P (1990) Effects of endogenous and exogenous cholesterol on the ultrastructure and steroid secretion of undifferentiated rat adrenocortical cells in primary culture. Cell Tissue Res 259:421–427

    Google Scholar 

  • Heikkilä P, Kahri AI, Ehnholm C, Kovanen PT (1988) Endogenous cholesterol synthesis is sufficient for ACTH-induced differentiation of rat adrenocortical cells in primary culture. In Vitro Cell Dev Biol 24:936–942

    Google Scholar 

  • Heikkilä P, Kahri AI, Enholm 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

    Google Scholar 

  • Hoerr N (1931) The cells of the suprarenal cortex in the guinea pig. Their reaction to injury and their replacement. Am J Anat 48:139–197

    Google Scholar 

  • Horiba N, Nomura K, Hizuka N, Takano K, Demura H, Shizume K (1987) Effects of IGF-I on proliferation and steroidogenesis of cultured adrenal zona glomerulosa cells. Endocrinol Jpn 34:611–614

    Google Scholar 

  • Hornsby PJ, Gill GN (1977) Hormonal control of adrenocortical cell proliferation. J Clin Invest 60:342–352

    Google Scholar 

  • Jackson S, Hodgkinson S, Estivariz FE, Lowry PJ (1991) IGF 1 and 2 in two models of adrenal growth. J Steroid Biochem Mol Biol 40:399–404

    Google Scholar 

  • Kahri AI (1966) Histochemical and electron microscopic studies on the cells of the rat adrenal cortex in tissue culture. Acta Endocrinol (Copenh) 108 [Suppl]:1–96

    Google Scholar 

  • Kahri AI (1973) Inhibition of ACTH-induced differentiation of cortical cells and their mitochondria by corticosterone in tissue culture of fetal rat adrenals. Anat Rec 176:253–272

    Google Scholar 

  • 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

    Google Scholar 

  • Kahri AI, Salmenperä M, Saure A (1976) Effects of 5-bromodeoxyuridine on the ACTH-dependent mitochondrial biogenesis in cortical cells of fetal rat adrenals in tissue culture. J Cell Biol 71:951–956

    Google Scholar 

  • 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

    Google Scholar 

  • Kitay J (1961) Sex differences in adrenal cortical secretion in the rat. Endocrinology 68:18–24

    Google Scholar 

  • Liddle GW, Island D, Meador CK (1962) Normal and abnormal regulation of corticotropin secretion in man. Recent Prog Horm Res 18:125–166

    Google Scholar 

  • Masui H, Garren LD (1970) On the mechanism of action of adrenocorticotropic hormone. Stimulation of deoxyribonucleic acid polymerase and thymidine kinase activities in adrenal glands. J Biol Chem 245:2627–2632

    Google Scholar 

  • Mitchell RM (1948) Histological changes and mitotic activity in the rat adrenal during postnatal development. Anat Rec 101:161–185

    Google Scholar 

  • Naaman E, Chatelain P, Saez JM, Durand P (1989) In vitro effect of insulin and insulin-like growth factor I on cell multiplication and adrenocorticotropin responsiveness of fetal adrenal cells. Biol Reprod 40:570–577

    Google Scholar 

  • Penhoat A, Chatelain PG, Jaillard C, Saez JM (1988) Characterization of insulin-like growth factor I and insulin receptors on cultured bovine adrenal fasciculata cells. Role of these peptides on adrenal cell function. Endocrinology 122:2518–2526

    Google Scholar 

  • Pillion DJ, Yang M, Grizzle WE (1988) Distribution of receptors for insulin and insulin-like growth factor I (somatomedin C) in the adrenal gland. Biochem Biophys Res Commun 154:138–145

    Google Scholar 

  • Pillion DJ, Arnold P, Yang M, Stochard CR, Grizzle WE (1989) Receptors for insulin and insulin-like growth factor I in the human adrenal gland. Biochem Biophys Res Commun 165:204–211

    Google Scholar 

  • Ramachandran J, Suyama AT (1975) Inhibition of replication of normal adrenocortical cells in culture by adrenocorticotropin. Proc Natl Acad Sci USA 72:113–117

    Google Scholar 

  • 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 (Copenh) 83:781–793

    Google Scholar 

  • Salmenperä M, Kahri AI (1977) Studies on the dependence of mitochondrial 11β- and 18-hydroxylation on the nuclear and mitochondrial DNA synthesis during ACTH-induced differentiation of cortical cells of rat adrenals in tissue culture. Exp Cell Res 104:223–232

    Google Scholar 

  • Salmenperä M, Kahri AI, Saure A (1976) Effects of corticosterone on adrenocorticotrophin-induced mitochondrial differentiation with special references to 11β- and 18-hydroxylation. J Endocrinol 70:215–222

    Google Scholar 

  • Shigematsu K, Niwa K, Kurihara M, Yamashita K, Kawai K, Tsuchiyama H (1989) Receptor autoradiographic localisation of insulin-like growth factor-I (IGF-I) binding sites in human fetal and adult adrenal glands. Life Sci 45:383–389

    Google Scholar 

  • Taylor JE, Scott CD, Baxter RC (1987) Comparison of receptors for insulin-like growth factor II from various tissues. J Endocrinol 115:35–41

    Google Scholar 

  • Van Dijk JP, Challis JRG, Tanswell AK (1988a) Basal serum-free culture system which supports growth of fetal rat adrenal cells in primary monolayer cell culture. J Dev Physiol 10:399–409

    Google Scholar 

  • Van Dijk JP, Tanswell AK, Challis JRG (1988b) Insulin-like growth factor (IGF)-II and insulin but not IGF-I, are mitogenic for fetal rat adrenal cells in vitro. J Endocrinol 119:509–516

    Google Scholar 

  • Voutilainen R, Miller WL (1987) Coordinate trophic hormone regulation of mRNAs for insulin-like growth factor II and the cholesterol side-chain-cleavage enzyme, P450scc, in human steroidogenic tissue. Proc Natl Acad Sci USA 84:1590–1594

    Google Scholar 

  • Voutilainen R, Miller WL (1988) Developmental and hormonal regulation of mRNAs for insulin-like growth factor II and steroidogenic enzymes in human fetal adrenals and gonads. DNA 7:9–15

    Google Scholar 

  • Walker BE, Rennels EG (1961) Adrenal cortical cell replacement in the mouse. Endocrinology 68:365–374

    Google Scholar 

  • Wright NA (1971) Cell proliferation in the prepubertal male rat adrenal cortex: an autoradiographic study. J Endocrinol 49:599–609

    Google Scholar 

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Arola, J., Heikkilä, P. & Kahri, A.I. Biphasic effect of ACTH on growth of rat adrenocortical cells in primary culture. Cell Tissue Res 271, 169–176 (1993). https://doi.org/10.1007/BF00297555

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  • DOI: https://doi.org/10.1007/BF00297555

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