scholarly journals Changes in the activities of the enzymes of urea synthesis caused by dexamethasone and dibutyryladenosine 3′:5′-cyclic monophosphate in foetal rat liver maintained in organ culture

1976 ◽  
Vol 160 (2) ◽  
pp. 159-162 ◽  
Author(s):  
E Edkins ◽  
N C R Rïhä
Keyword(s):  
Cyclic Amp ◽  
Organ Culture ◽  
Rat Liver ◽  
Urea Cycle ◽  
Defined Medium ◽  
Urea Synthesis ◽  
Dibutyryl Cyclic Amp ◽  
Carbamoyl Phosphate ◽  
Foetal Rat ◽  
Ornithine Transcarbamoylase

Liver explants from 19-day foetal rats were maintained in organ culture, in a defined medium, for up to 48h. Both 6-N,2′-O-dibutyryl cyclic AMP, in the presence of theophylline, and dexamethasone caused an increase in the activities of carbamoyl phosphate synthase, argininosuccinate synthetase, argininosuccinate lyase and arginase. These increases could be abolished by simultaneously incubating the explants with cycloheximide. No change in the activity of ornithine transcarbamoylase was found with either hormone. Previous work has shown that injection of corticosteroids into 19.5-day foetal rats in utero did not cause an increase in the arginine synthetase system. Present results suggest that this lack of effect is not due to any incompetence of the foetal rat liver at this stage to respond to this agent. The observations on ornithine transcarbamoylase activity suggest that this enzyme is induced in the liver of the perinatal rat by neither corticosteroids nor hormones acting via cyclic AMP, and it may be that all the enzymes of the urea cycle are induced physiologically by an agent or agents as yet unidentified.

scholarly journals Regulation of tyrosine aminotransferase in foetal rat liver

1980 ◽  
Vol 186 (2) ◽  
pp. 609-612 ◽  
Author(s):  
S M Andersson ◽  
N C Räihä ◽  
J J Ohisalo
Keyword(s):  
Enzyme Activity ◽  
Methyl Ester ◽  
Cyclic Amp ◽  
Organ Culture ◽  
Rat Liver ◽  
Tyrosine Aminotransferase ◽  
Dibutyryl Cyclic Amp ◽  
Adult Rat ◽  
Foetal Rat

A specific tyrosine aminotransferase, separate from the aspartate aminotransferases, is present in low concentration in foetal rat liver at the 21st day of gestation. Intraperitoneal injections of tyrosine methyl ester into the foetuses in utero increase the activity 2-fold, whereas glucose injections decrease it. Tyrosine, dexamethasone and dibutyryl cyclic AMP induce the enzyme activity in organ culture to the same extent as in adult rat liver in vivo.

scholarly journals The regulation of carbamoyl phosphate synthase activity in rat liver mitochondria

1976 ◽  
Vol 154 (2) ◽  
pp. 415-421 ◽  
Author(s):  
J D. McGivan ◽  
N M. Bradford ◽  
J Mendes-Mourão
Keyword(s):  
Protein Content ◽  
Nitrogen Source ◽  
Rat Liver ◽  
Liver Mitochondria ◽  
Rat Liver Mitochondria ◽  
Urea Synthesis ◽  
Carbamoyl Phosphate ◽  
Citrulline Synthesis ◽  
Ornithine Transcarbamoylase ◽  
Phosphate Synthase

The rate at which isolated rat liver mitochondria synthesized citrulline with NH4C1 as nitrogen source was markedly dependent on the protein content of the diet. 2. Citrulline synthesis was not rate-limited by substrate concentration, substrate transport or ornithine transcarbamoylase activity under the conditions used. 3. The intramitochondrial content of an activator of carbamoyl phosphate synthase, assumed to be N-acetyl-glutamate, varied markedly with dietary protein content. The variation in the concentration of this activator was sufficient to account for the observed variation in the rates of citrulline synthesis if this synthesis were rate-limited by the activity of carbamoyl phosphate synthase. 4. The rates of urea formation from NH4Cl as nitrogen source in isolated liver cells showed variations in response to diet that closely paralleled the variations in the rates of citrulline synthesis observed in isolated mitochondria. 5. These results are consistent with the postulate that when NH4Cl plus ornithine are present in an excess, the rate of urea synthesis is regulated at the level of carbamoyl phosphate synthase activity.

scholarly journals Influence of glucagon, 6-N,2′-O-dibutyryladenosine 3′:5′-cyclic monophosphate and triamcinolone on the arginine synthetase system in perinatal rat liver

1972 ◽  
Vol 126 (1) ◽  
pp. 89-98 ◽  
Author(s):  
A L. Schwartz
Keyword(s):  
Cyclic Amp ◽  
Rat Liver ◽  
Cyclic Nucleotide ◽  
Time Course ◽  
Fold Increase ◽  
Dibutyryl Cyclic Amp ◽  
System Activity ◽  
Cyclic Monophosphate ◽  
Foetal Rat ◽  
Postnatal Rats

1. The administration of triamcinolone (19–190μg/animal) to postnatal rats increased the arginine synthetase system activity 1.2–2.5-fold above control values 24h after exposure to the hormone. Cortisol (hydrocortisone), however, increased the arginine synthetase system activity only when larger (190μg/animal) or repeated daily doses were given. Glucagon (100μg/animal) stimulated arginine synthetase system activity only after the second postnatal day. None of these agents increased the activity in 19.5–21.5-day foetuses after intrauterine administration. 2. The viability of foetal rat liver explants maintained in organ culture for up to 54h was validated both by ultramicroscopic examination and by incorporation of radioactive leucine and orotic acid. 3. In organ cultures of foetal rat liver explants (18.5 days to term), triamcinolone (20μg/ml of medium) evoked a 2.8–4.3-fold increase after 24h of incubation. This increase was completely inhibited by actinomycin D (25μg/ml) or cycloheximide (10μg/ml). Cortisol (5–50μg/ml) or glucagon (0.067–67μg/ml) also increased the arginine synthetase system activity above the respective control values, but there was no increase in activity with insulin (0.05–0.25i.u./ml). 4. Maximum concentrations of glucagon (67μg/ml), dibutyryl cyclic AMP (6-N,2′-O-dibutyryladenosine 3′:5′-cyclic monophosphate) (0.1mm) and triamcinolone (20μg/ml) incubated for 24h with foetal rat liver explants each produced between a two-and three-fold increase in the activity of the arginine synthetase system. Combinations of maximum amounts of glucagon and the cyclic nucleotide did not produce a greater effect than either agent alone. However, the combination of dibutyryl cyclic AMP with triamcinolone appeared to produce somewhat less than additive effects. 5. The effects of the cyclic nucleotide and triamcinolone were evident after 12h of incubation and increased steadily throughout the 24h of observation. This time-course of increased enzyme activity is very much slower than that reported for the induction of other enzymes in explant cultures of foetal rat liver.

scholarly journals N-Acetylglutamate in rat liver during foetal development

1984 ◽  
Vol 222 (3) ◽  
pp. 837-838 ◽  
Author(s):  
M Van Dijk ◽  
P Lund
Keyword(s):  
Rat Liver ◽  
Urea Cycle ◽  
Developmental Pattern ◽  
Foetal Development ◽  
Tissue Content ◽  
Foetal Rat ◽  
Foetal Life

N-Acetylglutamate is present in foetal rat liver at 17 days' gestation. The tissue content (approx. 50 nmol/g wet wt.) remains constant during later foetal life. The appearance of N-acetylglutamate does not parallel the developmental pattern of the urea cycle.

TESTOSTERONE SECRETION BY FOETAL RAT TESTES IN VITRO

1976 ◽  
Vol 71 (2) ◽  
pp. 231-238 ◽  
Author(s):  
RÉGINE PICON
Keyword(s):  
Cyclic Amp ◽  
Synthetic Medium ◽  
Functional Differentiation ◽  
Dibutyryl Cyclic Amp ◽  
Testosterone Secretion ◽  
Testosterone Production ◽  
Foetal Rat

SUMMARY Testosterone secretion by foetal rat testes (13½–21½ days of gestation) explanted for 3 days in a synthetic medium was measured every 24 h by radioimmunoassay. During the first day of explantation, the foetal testis produced, respectively, 1013 ± 132, 8734 ± 1118, 9179 ± 2185 and 3886 ± 309 (s.e.m.) pg/testis when explanted at 14½, 16½, 18½ and 21½ days respectively. Testosterone production by 13½-day-old testes was not detectable on the first day of culture, but appeared on subsequent days. Daily testosterone secretion increased on the 2nd and 3rd days of culture in 14½-day-old testes and decreased in older stages. These results suggest that the functional differentiation of the testis is independent of stimulatory factors like gonadotrophins. Dibutyryl cyclic AMP was found to stimulate testosterone production significantly from 14½ days of gestation onwards.

scholarly journals Real-time study of the urea cycle using 15N n.m.r. in the isolated perfused rat liver

1992 ◽  
Vol 287 (3) ◽  
pp. 813-820 ◽  
Author(s):  
A Geissler ◽  
K Kanamori ◽  
B D Ross
Keyword(s):  
Rat Liver ◽  
Urea Cycle ◽  
Isolated Perfused Rat Liver ◽  
15N Recovery ◽  
Urea Synthesis ◽  
Perfused Liver ◽  
Biochemical Assays ◽  
Low Concentrations ◽  
15N Urea ◽  
Rate Limiting

1. Isolated rat liver was perfused with 10 mM-15NH4Cl, 5 mM-lactate and 1 mM-ornithine, or with 3 mM-[15N]alanine and 1 mM-ornithine, in haemoglobin-free medium. The liver was physiologically stable for over 3 h and synthesized urea at the rate of 1.15 mumol.min-1.g of liver-1 (15NH4(+)-perfused) or 0.41 mumol.min-1.g-1 ([15N]alanine-perfused). 2. The perfused liver was continuously monitored by 15N n.m.r. spectroscopy at 20.27 MHz for 15N. Well-resolved 15N resonances of precursors and intermediates of the urea cycle, present at tissue concentrations of 0.2-3.0 mumol/g, were observed from the intact liver in 5-40 min of acquisition. Key metabolites in liver extract and the final perfusion medium were analysed by n.m.r. and by biochemical assays to determine fractional 15N enrichment and the total 15N recovery. 3. In 15NH4(+)-perfused liver (n = 6), 15N incorporation into glutamate and alanine (1.0-1.3 mumol/g), as well as progressive formation of [15N2]urea, was observed during the first 2 h of perfusion. In the second and third hour, hepatic concentrations of [omega-15N]citrulline and [omega, omega'-15N]argininosuccinate increased to n.m.r.-detectable levels (0.3-0.9 mumol/g). The [15N]aspartate pool was large in the absence of added ornithine, but on its addition was rapidly incorporated into argininosuccinate (n = 3). 4. In [15N]alanine-perfused liver, major metabolites were [15N]glutamate, [gamma-15N]glutamine and [15N]urea. Urea-cycle intermediates were undetectable. 5. The results suggest that, in intact liver provided with excess ammonia, low concentrations of cytosolic argininosuccinate synthetase and argininosuccinate lyase limited the rate of metabolite flux in the urea cycle. By contrast, in alanine-perfused liver at a physiological rate of urea synthesis, mitochondrial carbamoylphosphate synthetase was rate-limiting. 6. The potential utility of 15N n.m.r. for study of metabolite channelling through urea-cycle enzymes in intact liver is discussed.

scholarly journals The effects of glucagon and insulin on adenosine 3′:5′-cyclic monophosphate concentrations in an organ culture of mature rat liver

1973 ◽  
Vol 132 (4) ◽  
pp. 765-773 ◽  
Author(s):  
Kenneth Siddle ◽  
Barbara Kane-Maguire ◽  
Anthony K. Campbell
Keyword(s):  
Cyclic Amp ◽  
Organ Culture ◽  
Rat Liver ◽  
Incubation Medium ◽  
Radioactive Tracer ◽  
Maximal Increase ◽  
Low Concentrations ◽  
Glucagon And Insulin ◽  
Millipore Filters ◽  
Glucagon Concentration

1. A modified radioimmunoassay for cyclic AMP was developed from the method of Steiner et al. (1969). Cyclic [3H]AMP was used as the radioactive tracer. Free and antibody-bound nucleotides were separated by adsorption of protein to Millipore filters. The assay was used to measure amounts of cyclic AMP down to 0.1pmol in 50μl. 2. The effect of glucagon on cyclic AMP content in pieces of mature rat liver maintained for 6 days in organ culture was studied. 3. Cyclic AMP content in the tissue reached a maximum in 5–15min and then decreased. This may have been partly due to an inhibitor of glucagon action formed in the tissue. Small amounts of cyclic AMP were released into the incubation medium. 4. The maximal increase in cyclic AMP content produced by glucagon decreased over 6 days in culture. However, liver pieces cultured for 2 and 6 days were more sensitive to low concentrations of glucagon than were fresh liver pieces. Glucagon concentrations for half-maximal effects were approx. 1μm and 0.05μm for fresh liver and 2-day cultured liver respectively. 5. Insulin (3.5μm) lowered the cyclic AMP content by 30% in the presence of a submaximal glucagon concentration in liver cultured for 2 days. No effect of insulin was demonstrated on fresh liver pieces. 6. Insulin and glucagon were rapidly destroyed by fresh liver pieces.

scholarly journals Factors that prevent the premature appearance of glucokinase in neonatal rat liver

1980 ◽  
Vol 186 (3) ◽  
pp. 817-826 ◽  
Author(s):  
M J O Wakelam ◽  
M B Allen ◽  
D G Walker
Keyword(s):  
Angiotensin Ii ◽  
Cyclic Amp ◽  
Insulin Release ◽  
Rat Liver ◽  
Neonatal Rat ◽  
Oral Glucose ◽  
Dibutyryl Cyclic Amp ◽  
Simultaneous Treatment ◽  
Physiological Factors ◽  
Glucose Administration

1. The physiological factors that prevent the precocious appearance of glucokinase activity in the 13-day-old rat that can be induced by oral glucose administration were explored. 2. Evidence is presented that the galactose component of milk sugar is inhibitory. In the absence of this inhibitory galactose, the amount of glucose necessary to effect appreciable induction is greater than that present in milk. 3. The induction is prevented both by administration of mannoheptulose, which inhibits insulin release, and by excess insulin; the amount of insulin available therefore seems to be critical. 4. The inhibition of induction by galactose does not appear to be via competition with glucose but by enhancing insulin release and thereby making this excessive. The relative amounts of glucose and insulin appear to be important in regulating glucokinase induction. 5. The precocious induction of glucokinase by glucose is inhibited by simultaneous treatment with approriate amounts of adrenaline, glucagon, dibutyryl cyclic AMP or isoprenaline but not by vasopressin or angiotensin II. 6. No single cause of glucokinase induction in neonatal rat liver can be recognized. The process is subject to regulation by many factors at a time subsequent to when competence to synthesize the enzyme has been established.

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