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J F Strauss

Publications and source records attributed to J F Strauss.

At least 145 records · Page 8Linked to original sources
PubMed1990

2-hydroxyestradiol enhanced progesterone production by porcine granulosa cells: dependence on de novo cholesterol synthesis and stimulation of cholesterol side-chain cleavage activity and cytochrome P450scc messenger ribonucleic acid levels.

2-Hydroxyestradiol (2-OH-E2) stimulates progestin secretion by granulosa cells, but the intracellular locus of the stimulatory effect has not been clarified. The objectives of the present studies were to 1) determine the role of de novo sterol synthesis in the effect of 2-OH-E2 on progestin biosynthesis, and 2) examine the effects of 2-OH-E2 on cholesterol side-chain cleavage (SCC) activity and the level of messenger RNA (mRNA) for P450scc. Inhibition of 3-hydroxy-3-methylglutaryl-coenzyme A reductase with lovastatin (5 micrograms/ml) or mevinolin (5 micrograms/ml) reduced FSH- and 2-OH-E2-stimulated (but not E2-stimulated) progesterone production. Mevalonate (20 mM) enhanced basal progesterone production and reversed the inhibitory effect of lovastatin but did not affect progesterone biosynthesis in the presence of 2-OH-E2. As an index of the activity of cholesterol SCC enzyme, granulosa cells were exposed to 25-hydroxycholesterol (10 micrograms/ml) for 24 h and progesterone secretion monitored. Conversion of 25-hydroxycholesterol into progesterone was stimulated 2- to 3-fold by maximally effective concentrations of 2-OH-E2, E2, and FSH. 2-OH-E2 and/or E2 further enhanced 25-hydroxycholesterol conversion in the presence of FSH, LH, and epinephrine. Aminoglutethimide, an inhibitor of SCC, reduced 2-OH-E2- and 2-OH-E2 plus FSH-stimulated progesterone production by 97% and 95%, respectively. 2-OH-E2 also increased basal (by 2 to 3-fold) and FSH-stimulated (to 3.5-fold of FSH-treated controls) levels of mRNA for cytochrome P450scc. Collectively, our studies support the hypothesis that 2-OH-E2-enhanced progesterone biosynthesis by porcine granulosa cells is dependent on de novo cholesterol synthesis and is associated with increased levels of the mRNA encoding cytochrome P-450scc, which leads to increases in basal and gonadotropin-induced SCC activity.

Aminoglutethimide↗
PubMed1990

Cloning and sequence of cDNA for human placental cytokeratin 8. Regulation of the mRNA in trophoblastic cells by cAMP.

A 1735 bp cDNA for human placental cytokeratin 8 is described which encompasses the entire coding sequence as well as 33 and 250 base pairs of 5'- and 3'-untranslated region, respectively. The level of cytokeratin 8 mRNA in various fetal tissues and placentae of different gestational ages was determined as were the effects of 8-bromo-cAMP on cytokeratin 8 mRNA in primary cultures of cytotrophoblasts and JEG-3 choriocarcinoma cells. Cytokeratin 8 mRNA was abundant in fetal small intestine, placenta, pancreas, lung, liver, and kidney. Levels of cytokeratin 8 mRNA in placenta increased slightly during pregnancy. 8-Bromo-cAMP suppressed cytokeratin 8 mRNA in primary cultures of cytotrophoblasts, whereas the cAMP analog increased mRNA levels in JEG-3 cells, revealing differential regulation of this mRNA in normal and transformed trophoblastic cells.

Amino Acid Sequence↗
PubMed1989

Regulation of the gene for estrogenic 17-ketosteroid reductase lying on chromosome 17cen----q25.

17-Ketosteroid reductase (17KSR), also known as 17 beta-hydroxysteroid dehydrogenase, catalyzes the reversible interconversion of estradiol to estrone and of androstenedione to testosterone. Using a recently cloned human placental 17KSR cDNA, we show that the 1.4-kilobase mRNA for this enzyme is detected only in tissues producing estrogens, and a 2.4-kilobase mRNA is detected in some estrogenic tissues and some androgenic tissues. This tissue distribution suggests that the interconversion of androstenedione and testosterone may be mediated by a different enzyme. Southern blotting studies show that the mRNA for this estrogenic 17KSR is encoded by two very similar genes localized to chromosome 17cen----q25 by analysis of DNA from mouse/human somatic hybrid cell lines. 8-Br-cAMP increases the abundance of estrogenic 17KSR mRNA as well as mRNAs for other steroidogenic enzymes in JEG-3 choriocarcinoma cells. By contrast, cAMP decreases estrogenic 17KSR mRNA in primary cultures of human cytotrophoblasts and human granulosa cells, a pattern of tropic regulation that differs from other steroidogenic enzyme mRNAs.

17-Hydroxysteroid Dehydrogenases↗
PubMed1989

Control of low density lipoprotein receptor gene promoter activity. Ketoconazole inhibits serum lipoprotein but not oxysterol suppression of gene transcription.

We have examined the effects of ketoconazole, a drug which inhibits enzymes involved in cholesterol biosynthesis and metabolism, on the suppressive effects of serum lipoproteins and 25-hydroxycholesterol on low density lipoprotein (LDL) receptor gene promoter activity. A LDL receptor promoter-chloramphenicol acetyltransferase (CAT) fusion gene construct (pLDLR-CAT 6500) was transfected into JEG-3 choriocarcinoma cells, and the transfected cells were cultured in the absence or presence of serum, LDL, or serum and 25-hydroxycholesterol. Serum, LDL, and serum + 25-hydroxycholesterol reduced chloramphenicol acetyltransferase activity in cells transfected with pLDLR-CAT 6500, whereas these treatments had no effect upon enzyme activity in cells transfected with a control construct (pSV2CAT). Ketoconazole (50 microM) overcame the effects of serum and LDL on suppression of pLDLR-CAT 6500 expression, but could not override the combination of serum + 25-hydroxycholesterol. Ketoconazole had no significant effect on expression of pSV2CAT. The drug inhibited cholesterol side chain cleavage enzyme in the cells, but appeared to have no impact on the ability of cells to take up LDL-carried lipids. Our observations are consistent with the idea that serum lipoprotein cholesterol is metabolized to an effector substance which acts to suppress LDL receptor gene transcription. The generation of this effector seems to be sensitive to ketoconazole.

Cell Line↗
PubMed1989

Hepatic lipase in the rat ovary. Ovaries cannot synthesize hepatic lipase but accumulate it from the circulation.

Hepatic lipase is proposed to have a role in steroidogenesis through its involvement in the metabolism of high density lipoproteins. We examined the activity, synthesis, distribution, and uptake of this enzyme and assessed the content of its mRNA in luteinized ovaries. We found that during peak steroidogenesis, ovaries of pregnant mare's serum gonadotropin-human chorionic gonadotropin-treated immature rats contained heparin-releasable hepatic lipase-like activity which was neutralized in a dose-dependent manner by purified antibodies to hepatic lipase isolated from post-heparin perfusates of rat livers. Quantitative immunoelectron microscopy revealed that ovarian hepatic lipase occurred along endothelial cells and was 3-fold more abundant in blood vessels of corpora lutea than those of stroma. However, hepatic lipase was not synthesized by the ovary since radiolabeled enzyme was not immunoisolated from the medium of dispersed luteinized granulosa cells incubated with [35S]methionine whereas it was present in the medium of control cells (hepatocytes). Similarly, hepatic lipase mRNA was detectable in liver but not ovaries or kidneys by Northern or slot blot analyses or by the polymerase chain reaction. Finally, 125I-labeled hepatic lipase injected into tail veins was quickly cleared from the systemic circulation, accumulating in liver, ovaries, kidneys, and spleen. Subsequent heparin injection caused rapid reappearance of radioactivity in the bloodstream and a marked decline of radiolabel in liver and ovaries but a modest decrease of that in kidneys and none in spleen. Exogenous 125I-bovine serum albumin also accumulated in all four organs but was not displaced from liver or ovaries by subsequent administration of heparin. Taken together, these data suggest that steroidogenically active ovaries possess but do not synthesize hepatic lipase. Instead, hepatic lipase originating elsewhere, presumably in the liver, is accumulated from the circulation at heparin-sensitive sites in ovarian blood vessels.

Animals↗
PubMed1989

Effects of 8-bromo-cAMP on expression of endocrine functions by cultured human trophoblast cells. Regulation of specific mRNAs.

There is little information on the molecular events underlying the effects of cAMP on human chorionic gonadotropin (hCG) and particularly steroidal hormone production in normal trophoblasts. We examined the effects of 8-bromo-cAMP on mRNAs encoding two components of the cholesterol side-chain cleavage system, cytochrome P-450scc and adrenodoxin, and the alpha- and beta-subunits of hCG in cultured cytotrophoblasts. cAMP caused an increase in all of these mRNAs within 24 h, whereas actin mRNA declined. alpha-hCG mRNA increased first, followed by adrenodoxin, beta-hCG and cytochrome P-450scc mRNAs. The effects of 8-bromo-cAMP on alpha- and beta-hCG, adrenodoxin, and cytochrome P-450scc mRNAs, in cytotrophoblasts and JEG-3 choriocarcinoma cells, required the catalytic unit of protein kinases since H-7, a kinase inhibitor, blocked the increase in the mRNAs and prevented the stimulation of hCG and progesterone secretion. 8-Bromo-cAMP promoted a rapid increase in alpha-hCG mRNA in cytotrophoblasts in the presence of cycloheximide, an inhibitor of protein synthesis. In cytotrophoblasts, cycloheximide reduced basal and 8-bromo-cAMP-stimulated adrenodoxin mRNA abundance. In contrast, basal and cAMP-stimulated adrenodoxin mRNA was augmented by cycloheximide in JEG-3 cells.(ABSTRACT TRUNCATED AT 250 WORDS)

8-Bromo Cyclic Adenosine Monophosphate↗
PubMed1989

Accumulation of colony-stimulating factor 1 in amniotic fluid during human pregnancy.

Colony-stimulating factor 1 is a hematopoietic growth factor that increases 1000-fold in the uteri of pregnant mice, and its receptor is abundantly expressed in the human placenta. The concentration of colony-stimulating factor 1 in amniotic fluid at 33 to 40 weeks (9.0 +/- 1.1 ng/ml) was twofold higher than that at 16 to 18 weeks gestation (4.1 +/- 0.5 ng/ml), whereas maternal serum colony-stimulating factor 1 levels did not rise significantly. Colony-stimulating factor 1 was detected in endometrial extracts from pregnant women and levels were higher than those in extracts from nonpregnant women.

Amniotic Fluid↗
PubMed1989

Control of low density lipoprotein receptor gene expression in steroidogenic cells.

Low density lipoprotein (LDL)-carried cholesterol is a primary substrate for steroid hormone synthesis by luteinized human granulosa cells. Chorionic gonadotropin and 8-bromo-cAMP both increase LDL receptor levels in granulosa cells by stimulating accumulation of the receptor mRNA. LDL and 25-hydroxycholesterol reduce LDL receptor expression, but this suppressive effect is partially overcome by 8-bromo-cAMP. Using fusion gene constructs containing the LDL receptor gene promoter transfected into JEG-3 cells, a cyclic AMP responsive enhancer could not be identified in the LDL receptor gene upstream promoter in transfection studies. We suggest that the LDL receptor gene in human steroidogenic cells is under negative control by a sterol effector, but that a cyclic AMP triggered process overcomes, to some extent, the sterol-mediated suppression. The detailed mechanisms by which sterol and cyclic AMP modulate LDL receptor gene expression remain to be elucidated.

Cholesterol, LDL↗
PubMed1989

Regulation of human trophoblast function by glucocorticoids: dexamethasone promotes increased secretion of chorionic gonadotropin.

Steroid hormones are thought to play a role in controlling placental endocrine function. Since maternal free cortisol levels increase during gestation, and glucocorticoid receptors have been identified in placental tissue, we examined the effects of glucocorticoids on the production of CG by cultured human cytotrophoblasts. Treatment of cytotrophoblasts with 1 microM dexamethasone increased CG secretion by 6- to 10-fold over a 72-h period, whereas progesterone (1 microM) had no effect. The stimulatory effects of dexamethasone were blocked by the glucocorticoid antagonist RU 486, indicating a requirement for the glucocorticoid receptor. Intracellular accumulation of the CG alpha-subunit in response to dexamethasone was demonstrated by immunocytochemistry, and Northern blot analyses revealed that dexamethasone treatment increases CG alpha- and beta-subunit mRNA levels. Dexamethasone also enhanced the stimulatory effects of 8-bromo-cAMP on CG secretion. We conclude that glucocorticoids as well as cAMP modulate human trophoblast endocrine functions.

8-Bromo Cyclic Adenosine Monophosphate↗
PubMed1989

Plasminogen activator inhibitor types 1 and 2 in human trophoblasts. PAI-1 is an immunocytochemical marker of invading trophoblasts.

Trophoblast implantation, vascular remodeling, and maintenance of intervillous blood flow may depend on the regulated production of proteolytic enzymes such as plasminogen activator (PA). Since the functional activity of plasminogen activators is determined not only by the quantity of protease but also by levels of specific plasminogen activator inhibitors (PAI), we examined trophoblasts both in vitro and in vivo for the presence of two PAIs, PAI-1 and PAI-2. Cytotrophoblasts were isolated from first trimester or term placentae, cultured, and immunocytochemically stained using specific anti-PAI antibodies. The antiserum against PAI-1 demonstrated prominent cell-surface staining and some cytoplasmic staining. The antiserum generated against PAI-2 revealed a cytoplasmic localization, with some trophoblasts staining intensely, whereas others had no apparent reactivity. We also found that cultured cytotrophoblasts contain the mRNAs for PAI-1 and PAI-2. Immunohistochemical analysis of tissue sections from 8-, 16-, and 40-week implantation sites using antisera against PAI-1 demonstrated weak staining of villous syncytiotrophoblasts but prominent cytoplasmic staining of trophoblasts invading the decidua and myometrium. Antisera against PAI-2 stained the cytoplasm of villous syncytiotrophoblasts, but no staining was evident in villous cytotrophoblasts or in invading trophoblasts. We conclude that 1) human trophoblasts can express both PAI-1 and PAI-2 in vitro and in vivo and 2) prominent PAI-1 immunostaining defines invading trophoblasts, whereas PAI-2 is the predominant PAI accumulated in villous syncytiotrophoblasts. Thus, the various trophoblast forms have distinctive patterns of PAI expression.

Cells, Cultured↗
PubMed1989

Regulation of side-chain cleavage enzyme and 3 beta-hydroxysteroid dehydrogenase by Ca2+ second messenger and protein kinase C systems in the placenta of the cow.

The steroidogenic activity of the bovine placenta is not modulated by cyclic nucleotide-mediated mechanisms. However, both translocation of intracellular Ca2+ and influx of extracellular Ca2+ activate the side-chain cleavage enzyme and 3 beta-hydroxysteroid dehydrogenase. Protein kinase C activation in concert with Ca2+ mobilization also activates the side-chain cleavage enzyme. Cholesterol availability is a rate-limiting factor. Using polyclonal antibodies against bovine adrenal cytochrome P-450scc, the presence of P-450scc was demonstrated in both placental and luteal tissues. The cytochrome P-450scc was then localized, using gold-staining electron microscopy, in the mononuclear cells but not the binuclear cells of the placentome. The results suggest that cholesterol is metabolized by the mononuclear cell to pregnenolone, where it is further metabolized to progesterone by the mononuclear and binuclear cells.

1-Methyl-3-isobutylxanthine↗
PubMed1989

Control of the steroidogenic machinery of the human trophoblast by cyclic AMP.

Human cytotrophoblasts express adenylate cyclase activity and possess membrane-bound regulatory proteins that bind guanine nucleotides (G proteins). Stimulation of the cyclase by forskolin or addition of 8-bromo-cAMP augments progesterone secretion by cultured cytotrophoblasts at least in part, by promoting accumulation of components of the cholesterol side-chain cleavage system. This is the consequence of increased synthesis of the proteins participating in steroidogenesis as a result of the 8-bromo-cAMP-provoked increase in their respective mRNAs. We propose that progesterone synthesis by cytotrophoblasts is up-regulated by cyclic AMP, which acts to increase expression of genes encoding the steroidogenic machinery. Paracrine or autocrine factors may initiate this cascade by stimulating the cytotrophoblast adenylate cyclase.

8-Bromo Cyclic Adenosine Monophosphate↗
PubMed1989

Characteristics of premenstrual syndrome.

To characterize the symptoms associated with premenstrual syndrome (PMS), the menstrual symptom calendars of 100 women diagnosed with PMS were factor-analyzed. Scores on 18 symptoms were recorded daily for one menstrual cycle prior to the women's first clinic visits. Two factors were found to predominate throughout the cycle: The first included emotional and behavioral symptoms and the second included physical and cognitive symptoms. The two factors were constant throughout all the phases of the menstrual cycle. Emotional symptoms clustered together throughout the menstrual cycle and were the dominant symptoms on factor I. However, they loaded (correlation of the variable with the factor) on factors I and II during the postmenstrual phase. Behavioral symptoms loaded on factor I throughout the menstrual cycle; however, they were prevalent predominantly in the later half of the intermenstrual and the premenstrual phases. Physical symptoms loaded predominantly on factor II and were less prevalent than the emotional symptoms. The cognitive symptoms loaded on factor II during the premenstrual phase. These findings are relevant for defining PMS criteria for women who seek treatment.

Adult↗
PubMed1988

8-bromoadenosine cyclic 3',5'-phosphate rapidly increases 3-hydroxy-3-methylglutaryl coenzyme A reductase mRNA in human granulosa cells: role of cellular sterol balance in controlling the response to tropic stimulation.

Exposure of cultured human granulosa cells to 8-bromoadenosine cyclic 3',5'-phosphate (8-bromo-cAMP) resulted in a rapid increase in the content of the mRNA for 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, a rate-limiting enzyme in the de novo synthesis of cholesterol. HMG-CoA reductase mRNA levels increased within 2 h of stimulation and remained elevated for at least 6 h. Treatment of granulosa cells with 25-hydroxycholesterol, a soluble cholesterol analogue, in combination with aminoglutethimide to block conversion of cellular sterols to pregnenolone, resulted in suppression of HMG-CoA reductase mRNA. When cells were stimulated with 8-bromo-cAMP in the presence of 25-hydroxycholesterol and aminoglutethimide, the increase in HMG-CoA reductase mRNA provoked by the tropic agent was markedly attenuated. This indicates that 8-bromo-cAMP raises HMG-CoA reductase mRNA levels indirectly by accelerating steroidogenesis and depleting cellular sterol pools, thus relieving sterol-mediated negative feedback of HMG-CoA reductase gene expression. 25-Hydroxycholesterol in the presence of aminoglutethimide suppressed low-density lipoprotein (LDL) receptor mRNA, but 8-bromo-cAMP effected a significant stimulation of LDL receptor mRNA levels when added with hydroxysterol and aminoglutethimide. These findings reveal differential regulation of HMG-CoA reductase and LDL receptor mRNAs in the presence of sterol negative feedback.

8-Bromo Cyclic Adenosine Monophosphate↗
PubMed1988

The upstream promoter of the human LDL receptor gene does not contain a cyclic AMP response element.

Fusion genes containing segments of the promoter region of the human LDL receptor gene and the coding sequence of the bacterial enzyme, chloramphenicol acetyltransferase (CAT), were introduced into JEG-3 human choriocarcinoma cells. Constructs containing 177 base pairs of 5'-flanking DNA (pLDLR-CAT 234) or 6500 base pairs (pLDLR-CAT 6500) promoted CAT activity in transient expression assays. Although both pLDLR-CAT 234 and pLDLR-CAT 6500 contain sequences related to the recently reported consensus sequence for cyclic AMP responsiveness, treatment of the transfected JEG-3 cells with 8-bromo-cAMP did not increase CAT activity. The cyclic AMP analog did, however, stimulate steroidogenesis and hCG secretion and increase CAT activity in cells transfected with p18X2SV1CAT, which contains two copies of an 18 base pair sequence corresponding to the cAMP-responsive element of the human alpha chorionic gonadotropin gene.

8-Bromo Cyclic Adenosine Monophosphate↗
PubMed1988

Biosynthesis and turnover of a 34-kDa protein growth factor in human cytotrophoblasts.

Recently we isolated a new protein growth factor of 34 kDa from synctial membranes of human placenta. In its polypeptide molecular mass, antigenic structure, receptor binding specificity and partial amino acid sequence, it is unlike several known growth factors, hormones and other proteins. Here we report studies on its biosynthesis and turnover in cultured cytotrophoblasts from term human placenta. Expression of the 34-kDa protein in these cells was studied by immunoprecipitation and Western blot analyses using a highly specific antibody. The experiments have produced the following results. a) Immunostaining and Western blot analyses have demonstrated the presence of immunoreactive 34-kDa protein in isolated cytotrophoblasts. The protein is present in both freshly isolated cells and in cells that have fused in culture to form multinuclear syncytiotrophoblasts. b) Trophoblastic biosynthesis of the protein has been demonstrated by in vitro translation of cellular mRNA and by metabolic labelling experiments with intact cells. c) Pulse-chase experiments show that biosynthesis of the protein does not involve any detectable precursors of higher or lower molecular mass. d) Studies on turnover indicate that the synthesized protein is unusually stable with a half-life of 50-70 h.

Cells, Cultured↗