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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 271 (1993), S. 169-176 
    ISSN: 1432-0878
    Keywords: Adrenal cortex ; Differentiation ; Tissue culture ; Proliferation ; Ultrastructure ; Steroids ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary 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.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Tissue cultures of fetal rat adrenals were used to study the effects of corticosterone on the ACTH-induced ultrastructural differentiation of cortical cells and their mitochondria. Corticosterone in dosages of 0.2, 2.0, 5.0, 10, and 20 μg/ml (corresponding to concentrations of 6 × 10-7, 6 × 10-6, 1.5 × 10-5, 3 × 10-5, and 6 × 10-5 molar) was added alone or together with 100 mU/ml of ACTH to the culture medium, daily from the sixteenth day of cultivation up to and including the twenty-first day. Corticosterone alone induced no ultrastructural changes in cortical cells. Corticosterone in concentrations of 6 × 10-7 to 3 × 10-5 M given with ACTH induced hypertrophy of Golgi apparatus. Corticosterone in concentrations of 6 × 10-5 M inhibited the ACTH-induced differentiation of cortical cells. However, the nuclear chromatin increased and Golgi apparatus was strikingly hypertrophied. Mitochondria often aggregated adjacent to the nuclear envelope but their ultrastructure remained undifferentiated with tubular or tubulovesicular cristae. Ribosomes appeared as single particles. A marked increase of smooth surfaced endoplasmic reticulum was noted also in cortical cells treated with 6 × 10-5 M of corticosterone.The present observations suggest that corticosterone acts as an intracellular inhibitor in cortical cells. It appears to inhibit cytoplasmic protein synthesis at the ribosomal level and prevents synthesis of cytoplasmic mitochondrial protein synthesis stimulating factor and the latter, in turn, inhibits the activation of mitochondrial protein synthesis. A new model is presented to explain the regulation of growth and secretion in the adrenal cortex.
    Additional Material: 1 Tab.
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  • 3
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Tissue cultures of fetal rat adrenals were used to study effects of cycloheximide on the ACTH-induced synthesis of mitochondrial inner membranes in the cortical cells. Cycloheximide alone added to the culture medium in concentrations of 100-0.25 μg/ml killed the cortical cells during six days of treatment. At the dosage level of 0.15 μg/ml/6 days, it induced a decrease in the size of mitochondria and an increase in the pleomorphism of mitochondria. Cycloheximide at a concentration of 0.015 μg/ml/6 days induced no change in the ultrastructure of cortical cells. When cycloheximide in concentrations of 0.15 μg/ml/6 days was added to the culture medium together with 100 mU/ml/6 days of ACTH, the ACTH-induced changes of mitochondrial inner membranes (formation of 600 Å vesicles) was completely inhibited. It also suppressed the ACTH-induced development of smooth surfaced endoplasmic reticulum, hypertrophy of Golgi apparatus, development of microvilli and accumulation of lipids. However, it had no effects on the ACTH-induced increase in the number of polysomes 01 the increase of heterochromatin in the nucleus. Cycloheximide (0.08 μg/ml/6 days) given together with 100 mU/ml/6 days of ACTH had incomplete inhibitory effects on the ACTH-induced differentiation of the cortical cells. Cycloheximide (0.015 μg/ml/6 days) given together with ACTH resulted in only slight inhibition of ACTH-induced ultrastructural differentiation of adrenal cortical cells in vitro.The present observations suggest that (1) the stimulatory effect of ACTH on mitochondrial protein synthesis is dependent upon nuclear control of protein synthesis; (2) a specific cytoplasmic mitochondrial protein synthesis stimulating factor in the cytoplasm of cortical cells is dependent on ribosomal protein synthesis and is a mediator of ACTH action on the mitochondrion; (3) despite their apparent autonomy, the mitochondria in cortical cells are kept under nuclear control; (4) the only direct locus for the trophic effect of ACTH is in the nucleus.
    Additional Material: 4 Tab.
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  • 4
    ISSN: 1432-0878
    Keywords: Adrenal cortex ; Differentiation ; Tissue culture ; Mevinolin ; Steroids ; Ultrastructure ; Rat (Sprague-Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: 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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  • 5
    ISSN: 1432-0878
    Keywords: Adrenal cortex ; Differentiation ; Tissue culture ; Steroids ; Ultrastructure ; Lipoproteins ; Rat (Sprague-Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary We studied the effects of lipoprotein-derived cholesterol on the ACTH-induced differentiation of cultured fetal rat adrenocortical cells. For this purpose human plasma high-density lipoprotein3 (HDL3) or low-density lipoprotein (LDL) was added to culture media devoid of cholesterol, and thereafter the morphological changes in cells were monitored and the amounts of steroids synthesized were measured. It could be demonstrated that, ultrastructurally, upon ACTH-stimulation the adrenocortical cells differentiated into fasciculata-like cells even in the absence of lipoproteins in the culture medium. The addition of either HDL3 or LDL caused an increase in the number and size of cytoplasmic lipid droplets suggesting uptake and deposition of lipoprotein-derived cholesterol into the differentiating cells. The amount of steroids secreted from cells differentiating in media devoid of cholesterol was only half that observed in cells differentiating in serum-supplemented medium. Addition of either HDL3 or LDL increased the ACTH-stimulated steroid synthesis to the levels observed in serum-supplemented medium. This study demonstrates that both HDL3 and LDL are able to provide cholesterol for steroid synthesis accompanying the ACTH-induced differentiation of fetal rat adrenocortical cells.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 0002-9106
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Tissue cultures of fetal rat adrenals were used to study effects of chloramphenicol on the ACTH-induced synthesis of mitochondrial inner membranes in the cortical cells. Chloramphenicol alone added to the culture medium in concentrations of 0.003, 0.03, 0.3, and 0.6 μ mole/ml/6 days induced no changes in the ultrastructure of cortical cells. Chloramphenicol in concentrations of 0.3 and 0.6 μ mole/ml/6 days given together with 100 mu/ml/6 days of ACTH inhibited completely the ACTH induced changes of mitochondrial inner membranes (formation of 600 Å vesicles). Chloramphenicol in concentrations of 0.003 μ mole/ml/6 days caused no inhibition of the ACTH effects. In concentration of 0.03 μ mole/ml/6 days chloramphenicol resulted in incomplete inhibition of ACTH-induced formation of mitochondrial vesicular cristae. None of these doses of chloramphenicol affected other ACTH-induced changes in the fine structure of the cells such as increase of smooth surfaced endoplasmic reticulum, hypertrophy of Golgi apparatus, and development of microvillous processes of plasma membranes. Chloramphenicol also caused no inhibition of the ACTH-induced accumulation of lipid in the cytoplasm. In cultivated cortical cells of Charles River strain albino rats small groups of annulated lamellae are commonly observed.The present observations suggest that: (1) the development of mitochondrial inner membranes is dependent, at least in part, on mitochondrial protein synthesis; (2) ACTH stimulation of mitochondrial protein synthesis in cortical cells is independent of ACTH-induced stimulation of nuclear-DNA-dependent protein synthesis; (3) doses of chloramphenicol which inhibit specialization of mitochondrial inner membranes of fetal adrenal cortical cells are comparable to those which inhibit protein synthesis in bacteria.
    Additional Material: 1 Tab.
    Type of Medium: Electronic Resource
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