Independence of the androgen-induced mRNA synthesis on exogenous glucose in the ventral prostate of the rat

1982 ◽  
Vol 101 (3) ◽  
pp. 472-480
Author(s):  
Anja Kuosa
Keyword(s):  
Protein Synthesis ◽  
Ventral Prostate ◽  
Experimental Conditions ◽  
Hormonal Stimulation ◽  
Electrophoretic Patterns ◽  
Exogenous Glucose ◽  
Polysomal Mrna ◽  
Mrna Content ◽  
Hormonal Activation

Abstract. Poly (A)-containing RNA species were isolated from prostatic polysomes after administration of testosterone to castrated rats. After exposure of prostatic tissue to radiolabelled uridine in vitro for 1 h the radioactivity of polysomal poly (A)-RNA increased at 6–12 h after testosterone treatment, which coincides with the hormonal activation of protein synthesis under the same experimental conditions. The increased radioactivity of polysomal mRNA was not due to changes in the uptake and phosphorylation or radiolabelled uridine or in the endogenous content of uridine nucleotides representing thus an increased mRNA content. Electrophoretic patterns of double-labelled polysomal mRNA failed to show detectable amounts of androgenspecific species. The larger part of increase in mRNA content thus consists of mRNAs common to prostatic cells of both testosterone-treated and non-treated castrated animals. The hormonal stimulation of polysomal poly (A)-RNA accumulation was not dependent upon exogenous glucose which is in contrast to protein synthesis. However, glucose was necessary for the hormonal activation of the phosphorylation of [3H]uridine leading ultimately to increased labelling of all RNA species. It is concluded that increased accumulation of polysomal mRNA may be necessary, but not sufficient for the hormonal activation of protein synthesis.

scholarly journals The control of the form and function of the ribosomes in androgen-dependent tissues by testosterone

1973 ◽  
Vol 134 (3) ◽  
pp. 795-805 ◽  
Author(s):  
W. I. P. Mainwaring ◽  
P. A. Wilce
Keyword(s):  
Protein Synthesis ◽  
Prostate Gland ◽  
Concomitant Administration ◽  
Ventral Prostate ◽  
Hormonal Stimulation ◽  
Pronounced Increase ◽  
Form And Function ◽  
Mandatory Requirement

1. The ribosome content of the rat ventral prostate gland is controlled by the concentrations of circulating androgens and the polyribosomal complement of the total population of ribosomes is acutely dependent on androgenic stimulation. After the administration of testosterone to castrated rats in vivo, there is a pronounced increase in the amounts of heavy (150–240S) polyribosomes. 2. These results are consistent with a pronounced increase in the mRNA and rRNA content of the prostate gland after the administration of testosterone in vivo. 3. From studies conducted both in vitro, the heavy prostate polyribosomes formed after androgenic stimulation are particularly active in protein synthesis. 4. The androgen-stimulated increase in the formation of prostate polyribosomes has a mandatory requirement for sustained RNA and protein synthesis. 5. Since the androgen-mediated increase in prostate polyribosomes may also be suppressed by the concomitant administration of certain anti-androgenic steroids in vivo, the response in polyribosome formation is probably initiated by the binding of a metabolite of testosterone, 5α-dihydrotestosterone, in the prostate gland. 6. The relevance of these findings to the pronounced increase in protein synthesis in androgen-dependent tissues after hormonal stimulation is discussed.

Effects of prolactin on testosterone-induced growth and protein synthesis in rat accessory sex glands

1983 ◽  
Vol 96 (3) ◽  
pp. 407-416 ◽  
Author(s):  
R. Jones ◽  
P. R. Riding ◽  
M. G. Parker
Keyword(s):  
Protein Synthesis ◽  
Total Protein ◽  
Sodium Dodecyl ◽  
Chronic Administration ◽  
Specific Protein ◽  
Ventral Prostate ◽  
Plasma Prolactin ◽  
Accessory Sex Glands ◽  
Caput Epididymidis

The relative importance of testosterone and prolactin in regulating growth and protein synthesis in rat accessory sex glands has been investigated. Protein synthesis was measured by incubating tissue minces in vitro with [35S]methionine and analysing labelled proteins on polyacrylamide gels containing sodium dodecyl sulphate. Plasma prolactin was assayed by radioimmunoassay. Results showed that castration for 8 days significantly reduced wet weights and total protein synthesis in the ventral prostate, dorsolateral prostate and caput epididymidis, but that these effects could be reversed by exogenous testosterone. Similarly, the specific incorporation of [35S]methionine into four polypeptides in the ventral prostate, two polypeptides in the dorsolateral prostate and two polypeptides in the caput epididymidis was lowered by castration but markedly stimulated by testosterone. Acute or chronic administration of 2-bromo-α-ergocryptine to animals in combination with testosterone had no significant effect on any of the parameters measured, although the drug reduced circulating prolactin to undetectable levels. In addition, exogenous prolactin given alone, or in combination with testosterone, to hypophysectomized rats had no effect on general or specific protein synthesis. The induction of hyperprolactinaemia in immature or mature rats with pituitary homographs had no effect on testosterone-stimulated growth of any accessory gland, although it caused a significant stimulation of total protein synthesis in the dorsolateral prostate and coagulating glands. However, this was a generalized effect as it did not increase the specific incorporation of [35S]methionine into androgen-dependent proteins. The results do not indicate a major role for prolactin in regulating androgen responsiveness of male accessory sex glands in the rat.

scholarly journals Studies on the form and synthesis of messenger ribonucleic acid in the rat ventral prostate gland, including its tissue-specific stimulation by androgens

1974 ◽  
Vol 137 (3) ◽  
pp. 513-524 ◽  
Author(s):  
W. Ian P. Mainwaring ◽  
Peter A. Wilce ◽  
Allan E. Smith
Keyword(s):  
Prostate Gland ◽  
Sodium Dodecyl ◽  
Ventral Prostate ◽  
Experimental Conditions ◽  
Free System ◽  
Cell Free System ◽  
Messenger Ribonucleic Acid ◽  
Selective Incorporation

1. When prostate polyribosomes are labelled with radioactive precursors in vivo and subsequently dissociated with sodium dodecyl sulphate, a heterogeneous 6–15S RNA species may be identified that possesses all of the distinctive properties of mRNA. 2. Apart from the selective incorporation of 5′-fluoro-orotic acid into this 6–15S RNA component, it is bound by nitrocellulose filters under experimental conditions where only poly(A)-rich species of RNA are specifically retained. Most importantly, however, only the 6–15S RNA fraction is capable of promoting the incorporation of amino acids into peptide linkage in an mRNA-depleted cell-free system derived from ascites-tumour cells. 3. With the development of a simpler method for labelling the total RNA fraction of the prostate gland in vitro, the poly(A)-enriched RNA fraction may be readily isolated by adsorption and elution from oligo(dT)-cellulose. The synthesis of the poly(A)-enriched 6–15S RNA fraction is stringently controlled by androgens in a highly tissue- and steroid-specific manner. 4. From an analysis of the proteins synthesized in the ascites cell-free system in the presence of the poly(A)-rich RNA fraction, it appears that protein synthesis in the prostate gland is stimulated in a rather general way, even during the earliest phases of the androgenic response. This conclusion may require modification when more specific means of analysis are available than those used in the present investigation. 5. The implications of these findings to the mechanism of action of androgens are discussed.

A novel mechanism of olanzapine-induced lipid accumulation

2011 ◽  
Vol 26 (S2) ◽  
pp. 906-906 ◽  
Author(s):  
S. Dzitoyeva ◽  
H. Chen ◽  
R. Manev ◽  
H. Manev
Keyword(s):  
Lipid Content ◽  
Direct Action ◽  
Inhibitory Effect ◽  
Experimental Conditions ◽  
Cell Content ◽  
Metabolic Alterations ◽  
Low Concentrations ◽  
Mrna Content ◽  
Quantitative Western Blot

IntroductionSecond generation antipsychotic drugs (SGADs) including olanzapine trigger adverse metabolic alterations possibly by a direct action on adipocytes.Objectives and aimsThe system of the inflammatory 5-lipoxygenase (5-LOX) and its activating protein (FLAP) have been implicated in lipid dysfunction in obesity. We investigated whether this system could participate in the adipogenic action of olanzapine.MethodsExperiments were performed in 3T3-L1 adipocytes in vitro. Cells were treated with olanzapine and a FLAP inhibitor MK-886. Their lipid content, 5-LOX and FLAP mRNA content, and FLAP protein content were measured.ResultsOlanzapine treatment did not affect the cell content of 5-LOX mRNA; however, it decreased FLAP mRNA content at day five but not 24 hours after olanzapine addition. The inhibitory effect of olanzapine on FLAP expression was confirmed by quantitative Western blot assays. In the absence of a FLAP inhibitor, low concentrations of olanzapine (0.5 and 5 μM) increased lipid content only by about 13% (compared to about a 56% increase induced by 50 μM olanzapine) whereas in the presence of MK-886 these concentrations of olanzapine produced lipid increases comparable to the increase caused by 50 μM. In these experimental conditions, MK-886 alone did not alter the cell content of lipids.Conclusions5-LOX system may be involved in lipid dysfunction not only in conditions of obesity but possibly in SGAD-related metabolic alterations. The known polymorphism in the genes of the human 5-LOX system could play a role in setting a variable individual susceptibility to the metabolic side effects of SGADs.

scholarly journals Hormonal stimulation in the exocrine pancreas results in coordinate and anticoordinate regulation of protein synthesis.

1984 ◽  
Vol 99 (5) ◽  
pp. 1569-1574 ◽  
Author(s):  
J Schick ◽  
H Kern ◽  
G Scheele
Keyword(s):  
Protein Synthesis ◽  
Fold Increase ◽  
Response Patterns ◽  
Hormonal Stimulation ◽  
Group Iii ◽  
Hormone Stimulation ◽  
Control Value ◽  
Group I ◽  
Separation Of Proteins

24-h intravenous caerulein infusion studies in the rat were combined with in vitro amino acid incorporation studies followed by high-resolution separation of proteins by two-dimensional isoelectric focusing and SDS gel electrophoresis to study the extent to which persistent changes in the biosynthesis of exocrine pancreatic proteins are regulated by cholecystokinin-like peptides. Beginning in the third hour of optimal hormone infusion at 0.25 microgram kg-1 h-1, changes were observed in the synthetic rates of 12 proteins, which progressed over the course of the 24-h study. Based on coordinate response patterns, exocrine proteins could be classified into four distinct groups. Group I (trypsinogen forms 1 and 2) showed progressive increases in synthetic rates reaching a combined 4.3-fold increase over control levels. Group II (amylase forms 1 and 2) showed progressive decreases in synthesis to levels 7.1- and 14.3-fold lower than control levels, respectively. Group III proteins (ribonuclease, chymotrypsinogen forms 1 and 2, procarboxypeptidase forms A and B, and proelastase 1) showed moderate increases in synthesis, 1.4-2.8-fold, and group IV proteins (trypsinogen 3, lipase, proelastase 2, and unidentified proteins 1-4) did not show changes in synthesis with hormone stimulation. Regulation of protein synthesis in response to caerulein infusion was specific for individual isoenzymic forms in the case of both trypsinogen and proelastase. The ratio of biosynthetic rates of trypsinogen forms 1 + 2 to amylase forms 1 + 2 increased from a control value of 0.56 to 24.4 after 24 h of hormonal stimulation (43.5-fold increase). Biosynthetic rates for an unidentified protein (P23) with an Mr = 23,000 and isoelectric point of 6.2 increased 14.2-fold, and the ratio of synthesis of P23 to amylase 2 increased 200-fold during caerulein infusion. During hormone stimulation the anticoordinate response in the synthesis of pancreatic glycosidases (decreased synthesis) and serine protease zymogens (increased synthesis) explain previous observations that showed little change in rates of total protein synthesis under similar conditions.

Early Dissociation of Protein Synthesis and Amylase Secretion Following Hormonal Stimulation of the Pancreas

1974 ◽  
Vol 52 (2) ◽  
pp. 198-205 ◽  
Author(s):  
R. Mongeau ◽  
Y. Couture ◽  
J. Dunnigan ◽  
J. Morisset
Keyword(s):  
Protein Synthesis ◽  
Incubation Medium ◽  
Single Injection ◽  
Pancreatic Enzyme ◽  
Amylase Secretion ◽  
Hormonal Stimulation ◽  
Pancreatic Enzyme Secretion ◽  
Metabolic Conditions

The secretion of the various pancreatic enzymes can be increased by hormonal and cholinergic stimulation. However, it is not yet clear among the different investigators if their synthesis remains constant or can be modified according to different metabolic conditions. The secretion and synthesis of the pancreatic proteins were then studied in parallel to evaluate if secretion triggers synthesis or both phenomenons are controlled by separate mechanisms.The approach for these studies consists mainly in a combination of in vivo and in vitro experiments. The stimulants were injected in vivo and the pancreatic secretions were collected for different periods of time. The animals were then sacrificed and protein synthesis was measured in vitro along with the amylase secreted into the incubation medium. The results show that protein synthesis is decreased during the first 15 min after a single injection or infusion of both cholecystokinin–pancreozymin (CCK–PZ) and secretin. This reduction was associated with an increase in amylase secreted into the incubation medium. However, at 30 min after the hormonal stimulation, protein synthesis is increased while secretion into the incubation medium had returned to control levels. This increase in protein synthesis lasts for at least 1 h. These results strongly suggest that pancreatic enzyme secretion and synthesis are dissociated in the early minutes following hormonal stimulation.

Influence of steroid hormones on the incorporation of amino acids in uterine and cervical tissue of pregnant women

1987 ◽  
Vol 115 (4) ◽  
pp. 537-543
Author(s):  
Ingrid Wiqvist ◽  
Anders Linde
Keyword(s):  
Amino Acids ◽  
Protein Synthesis ◽  
Pregnant Women ◽  
Tissue Concentration ◽  
Uterine Tissue ◽  
Protein Synthesis Inhibitors ◽  
Experimental Conditions ◽  
Pregnant Uterus ◽  
High Concentrations

Abstract. The influence of steroids on protein synthesis in cervical and uterine tissue obtained from early and term pregnant women was studied by measuring the incorporation of labelled amino acids into total protein. It was found that oestradiol-17β and progesterone significantly reduced the incorporation of [3H]proline. Androstenedione and cortisol had no significant effect on the incorporation of [3H]proline even at high concentrations. The protein synthesis inhibitors puromycin and cycloheximide blocked the incorporation of [3H]proline to 80–85%. However, there was no further reduction in the incorporation in the presence of oestradiol. Oestradiol was found to reduce the incorporation of [14C]glycine but not that of [3H]serine. The results indicate that oestradiol and progesterone reduce protein synthesis in human cervical and uterine tissue and that this reduction, at least partially, involves collagen synthesis. Oestradiol and progesterone were equipotent under in vitro experimental conditions. The tissue concentration of progesterone in the pregnant uterus is, however, much higher than that of oestradiol. It seems therefore probable that progesterone rather than oestradiol restricts unopposed synthesis of proteins, presumably mainly collagen.

scholarly journals Specific changes in the messenger ribonucleic acid content of the rat ventral prostate gland after androgenic stimulation. Evidence from the synthesis of aldolase messenger ribonucleic acid

1974 ◽  
Vol 144 (2) ◽  
pp. 413-426 ◽  
Author(s):  
W I P Mainwaring ◽  
F R Mangan ◽  
R A Irving ◽  
D A Jones
Keyword(s):  
Ribonucleic Acid ◽  
De Novo ◽  
Prostate Gland ◽  
Ventral Prostate ◽  
Mrna Synthesis ◽  
Hormonal Stimulation ◽  
Messenger Ribonucleic Acid ◽  
Stimulation Of

1. Aldolase was selected as a suitable marker for following the androgenic regulation of mRNA synthesis in the prostate gland. 2. Antibodies raised in rabbits against crystalline prostate aldolase were used to monitor the synthesis of this androgen-induced enzyme after hormonal stimulation of castrated animals, by using procedures in vivo and in vitro for the translation of prostate poly(A)-rich mRNA. 3. After androgenic stimulation in vivo the poly(A)-rich mRNA was isolated from the prostate gland and other tissues of castrated rats, and added to a protein-synthesizing system in vitro derived from Krebs II ascites-tumour cells. By using this approach it was found that androgens regulate the synthesis of aldolase mRNA in a highly tissue-specific manner. Stimulation of aldolase mRNA synthesis reached a maximum after 8h of androgenic treatment and then declined. 4. The androgenic control of aldolase mRNA synthesis was also investigated in vivo. After treatment of castrated animals with various steroids in vivo [35S]methionine was injected directly into the prostate gland, and labelled aldolase was selectively precipitated from isolated polyribosomes with anti-aldolase serum. The regulation of aldolase mRNA synthesis in the prostate gland was stringently steroid-specific and could only be evoked by androgens. After a single injection of testosterone, aldolase synthesis reached a maximum after 16h of hormonal stimulation and then declined. 5. Although androgens exert significant control over transcriptional processes in the prostate gland, and appear to regulate the synthesis of aldolase mRNA de novo, the possibility exists for additional means of control at the translational level of aldolase synthesis. The results are discussed in the context of the overall mechanism of action of androgens.

3α-HYDROXYSTEROID DEHYDROGENATION IN SUBCELLULAR FRACTIONS OF RAT VENTRAL PROSTATE AND OTHER TISSUES

1965 ◽  
Vol 33 (3) ◽  
pp. 353-363 ◽  
Author(s):  
MARION B. R. GORE ◽  
D. N. BARON
Keyword(s):  
Seminal Vesicle ◽  
Chromatographic Method ◽  
Ventral Prostate ◽  
Rat Tissues ◽  
Soluble Enzyme ◽  
Experimental Conditions ◽  
Widespread Occurrence ◽  
Rat Ventral Prostate ◽  
Soluble Fractions

SUMMARY Dehydrogenation of androsterone, catalysed by both particulate and soluble fractions of rat ventral prostate, has been demonstrated in vitro by spectrophotometric and chromatographic methods. A difference has been observed between dehydrogenases in the particulate and soluble fractions in their acceptance, under uniform experimental conditions, of the 5α and 5β isomers, androsterone and aetiocholanolone, as substrate. The particulate enzyme dehydrogenates androsterone under conditions in which aetiocholanolone is not dehydrogenated, whereas the soluble enzyme utilizes equally androsterone and aetiocholanolone as substrate. An examination of other rat tissues by the chromatographic method has confirmed the widespread occurrence of the soluble dehydrogenase. Particulate dehydrogenase resembling the prostatic enzyme was detected in seminal vesicle and kidney.

scholarly journals Deriving accurate molecular indicators of protein synthesis through Raman-based sparse classification

2021 ◽  
Author(s):  
N. Pavillon ◽  
N. I. Smith
Keyword(s):  
Protein Synthesis ◽  
Linear Models ◽  
Linear Method ◽  
Classification Performance ◽  
Experimental Conditions ◽  
Protein Levels ◽  
Spectral Coefficients ◽  
Rna Accumulation ◽  
Molecular Indicators

AbstractRaman spectroscopy has the ability to retrieve molecular information from live biological samples non-invasively through optical means. Coupled with machine learning, it is possible to use the large amount of information contained in a Raman spectrum to create models that can predict the state of new samples based on statistical analysis from previous measurements. Furthermore, in case of linear models, the separation coefficients can be used to interpret which bands are contributing to the discrimination between experimental conditions, which correspond here to single-cell measurements of macrophages under in vitro immune stimulation. We here evaluate a typical linear method using discriminant analysis and PCA, and compare it to regularized logistic regression (Lasso). We find that the use of PCA is not beneficial to the classification performance. Furthermore, the Lasso approach yields sparse separation vectors, since it suppresses spectral coefficients which do not improve classification, making interpretation easier. To further evaluate the approach, we apply the Lasso technique to a well-defined case where protein synthesis is inhibited, and show that the separating features are consistent with RNA accumulation and protein levels depletion. Surprisingly, when Raman features are selected purely in terms of their classification power (Lasso), the selected coefficients are contained in side bands, while typical strong Raman peaks are not present in the discrimination vector. We propose that this occurs because large Raman bands are representative of a wide variety of cellular molecules and are therefore less suited for accurate classification.

Sign in / Sign up

Export Citation Format

Share Document