scholarly journals Gluconeogenesis from glycine and serine in fasted normal and diabetic rats

1988 ◽  
Vol 253 (1) ◽  
pp. 27-32 ◽  
Author(s):  
G Hetenyi ◽  
P J Anderson ◽  
M Raman ◽  
C Ferrarotto

1. Non-anaesthetized normal and diabetic rats were fasted for 1 day, and [U-14C]glycine, or [U-14C]serine, or [U-14C]- plus [3-3H]-glucose was injected intra-arterially. The rates of synthesis de novo/irreversible disposal for glycine, serine and glucose, as well as the contribution of carbon atoms by the amino acids to plasma glucose, were calculated from the integrals of the specific-radioactivity-versus-time curves in plasma. 2. The concentrations of both glycine and serine in blood plasma were lower in diabetic than in fasted normal animals. 3. The rates of synthesis de novo/irreversible disposal of both amino acids tended to be lower in diabetic animals, but the decrease was statistically significant only for serine (14.3 compared with 10.5 mumol/min per kg). 4. Of the carbon atoms of plasma glucose, 2.9% arose from glycine in both fasted normal and diabetic rats, whereas 4.46% of glucose carbon originated from serine in fasted normal and 6.77% in diabetic rats. 5. As judged by their specific radioactivities, plasma serine and glycine exchange carbon atoms rapidly and extensively. 6. It was concluded that the turnover of glycine remains essentially unchanged, whereas that of serine is decreased in diabetic as compared with fasted normal rats. The plasma concentration of both amino acids was lower in diabetic rats. Both glycine and serine are glucogenic. In diabetic rats the contribution of carbon atoms from glycine to glucose increases in direct proportion to the increased glucose turnover, whereas the contribution by serine becomes also proportionally higher.

1971 ◽  
Vol 50 (1) ◽  
pp. 187-200 ◽  
Author(s):  
Abraham Amsterdam ◽  
Michael Schramm ◽  
Itzhak Ohad ◽  
Yoram Salomon ◽  
Zvi Selinger

After enzyme secretion the membrane of the secretory granule, which had been fused to the cell membrane, was resorbed into the cell. Experiments were therefore carried out to test whether formation of new secretory granules involves reutilization of the resorbed membrane or synthesis of a new membrane, de novo, from amino acids. Incorporation of amino acids-14C into proteins of various cell fractions was measured in vivo, 30, 120, and. 300 min after labeling. At all times the specific radioactivity of the secretory granule membrane was about equal to that of the granule's exportable content. At 120 and 300 min the specific radioactivity of the granule membrane and of the granule content was much higher than that of any other subcellular fraction. It is therefore concluded that the protein of the membrane is synthesized de novo concomitantly with the exportable protein. The proteins of the granule membrane could be distinguished from those of the granule content by gel electrophoresis. All major bands were labeled proportionately to their staining intensity. The amino acid composition of the secretory granule membrane was markedly different from that of the granule's content and also from that of the mitochondrial membrane. The granule membrane showed a high proline content, 30 moles/100 moles amino acids. The analyses show that the radioactivity of the granule membrane is indeed inherent in its proteins and is not due to contamination by other fractions. The possibility is considered that the exportable protein leaves the endoplasmic reticulum already enveloped by the newly synthesized membrane.


1986 ◽  
Vol 251 (1) ◽  
pp. E71-E77 ◽  
Author(s):  
S. C. Denne ◽  
S. C. Kalhan

Total carbohydrate oxidation, plasma glucose oxidation, and glucose carbon recycling were measured in 11 fasting newborns using a constant infusion of D-[U-13C]glucose combined with respiratory calorimetry. The "true" rate of glucose appearance (Ra) was quantified from the enrichment of the nonrecycling tracer species (m + 6), while the "apparent" rate of glucose appearance was quantified from the enrichment of glucose C - 1. The plasma glucose concentration remained constant at approximately 50 mg/dl (2.8 mM) throughout the study. The true rate of glucose production was 5.02 +/- 0.41 mg X kg-1 X min-1, (means +/- SD). Glucose was oxidized at a rate of 2.67 +/- 0.34 mg X kg-1 X min-1 and represented 53% of the glucose turnover. Recycling of glucose carbon represented 36% of the glucose production rate, or 1.87 +/- 0.74 mg X kg-1 X min-1. The oxidation of plasma glucose provided 15.8 +/- 2.0 kcal X kg-1 X day-1, whereas total carbohydrate oxidation (measured by respiratory calorimetry) provided 19.9 +/- 6.6 kcal X kg X day. The data indicate that 1) recycling of glucose carbon accounts for about one-third of glucose production, demonstrating active gluconeogenesis in the fasting newborn; 2) the oxidation of plasma glucose represents only 80% of total carbohydrate oxidation, the remaining 20% possibly representing the local oxidation of tissue glycogen stores; and 3) as the measured rate of glucose oxidation will be insufficient to supply the entire calculated cerebral metabolic requirements, these data suggest that fuels in addition to glucose may be important for cerebral metabolism in the fasting human newborn.


1984 ◽  
Vol 224 (2) ◽  
pp. 355-360 ◽  
Author(s):  
G Hetenyi ◽  
P J Anderson ◽  
G A Kinson

L-[U-14C]Threonine was infused at a steady rate to non-anaesthetized rats starved for 1 or 3 days and to diabetic rats starved for 1 day. The rates of turnover of threonine, calculated from the equilibrium specific radioactivity (SA) of plasma threonine, were 5.79 +/- 1.00, 11.67 +/- 1.43 and 13.35 +/- 1.85 mumol/min per kg body wt. in 1-day-starved, 3-day-starved and diabetic rats respectively. The calculated turnover rate of threonine agreed well with the rate expected from the rate of protein turnover reported in the literature. The equilibrium SA of plasma alanine was 5.1-9.8% of that of threonine in the three groups of rats. The equilibrium SA of glucose was 1.42 and 2.90% of that of threonine in 1-day- and 3-day-starved rats respectively. From the non-equilibrium SA of glucose, it is estimated that a higher percentage of 14C atoms is transferred from threonine to glucose in diabetic than in non-diabetic rats. In spite of increases in gluconeogenesis from threonine in long-starved or diabetic rats, we conclude that threonine remains a minor contributor to plasma glucose. Since it is an essential amino acid, its turnover and contribution to the formation of plasma glucose is an index of catabolism and gluconeogenesis from tissue protein.


1980 ◽  
Vol 58 (7) ◽  
pp. 787-796 ◽  
Author(s):  
G. Hetenyi Jr. ◽  
B. Pagurek ◽  
Elizabeth A. Dittmar ◽  
Catherine Ferrarotto

The turnover rate of glucose, the irreversible disposal rate of alanine, and the transfer of carbon atoms from alanine to glucose were studied with the help of tracer methods in normal, methylprednisolone (MP) treated and pancreatectomized dogs. The incorporation of carbon from alanine into pyruvate was also followed. In comparison with normal dogs, in MP-treated dogs the plasma level of alanine and pyruvate were elevated by a factor of about 3 whereas the level of plasma glucose was increased by 10%. Both the turnover rate of glucose and the irreversible disposal rate of alanine were significantly higher. In pancreatectomized dogs plasma alanine and pyruvate levels were not different from normal. There was no increase in the irreversible disposal rate of alanine. Glucose turnover was about twice as much as in normals. The transfer of C atoms from alanine to glucose was increased in both MP-treated and pancreatectomized dogs. The increase was in direct proportion to the increase in the irreversible disposal rate of alanine in MP-treated dogs whereas in pancreatectomized dogs it was due to an increased fraction of the irreversible disposal rate of alanine being converted to circulating glucose. Thus two different mechanisms are responsible for the increased carbon transfer and hence presumably gluconeogenesis in MP-treated and in pancreatectomized animals. The role of insulin as the determinant of the preferred route to increase gluconeogenesis is discussed.


2014 ◽  
Vol 54 (10) ◽  
pp. 1728 ◽  
Author(s):  
M. Al-Mamun ◽  
Y. Sako ◽  
H. Sano

Isotopic dilution methodology of [U-13C]glucose and [1-13C]Na-acetate were performed simultaneously as primed continuous infusions to determine plasma glucose and acetate metabolism in sheep during thermoneutral (TN) and cold exposure (CE, 2−4°C). The experiment was performed using crossbred (Corriedale × Suffolk) sheep (n = 5; 3 male and 2 female; ~2 years old, 38 ± 2.5 kg of initial bodyweight). The animals were offered mixed hay of orchardgrass and reed canarygrass (40 : 60) 62 g/kg0.75/day with ad libitum water access. Concentration of plasma glucose was determined enzymatically using the glucose oxidase method. Plasma concentration of non-esterified fatty acids (NEFA) and glucose were higher (P = 0.01 and P = 0.05 respectively) during CE than TN. Plasma concentration and the turnover rate of acetate were numerically higher (P = 0.09 and P = 0.25 respectively) during CE than during TN. Plasma glucose turnover rate was higher (P = 0.02) during CE than TN. The present findings suggested that plasma acetate turnover rate tended to be elevated during CE, with further more extensive studies required to clarify the significance.


1977 ◽  
Vol 162 (3) ◽  
pp. 643-651 ◽  
Author(s):  
D F Heath ◽  
K N Frayn ◽  
J G Rose

1. Rates and rate coefficients of glucose utilization and replacement were determined with [5-3H]- and [U-14C]-glucose in rats starved for 24h, either conscious or under halothane anaesthesia, in a thermoneutral environment. Plasma insulin concentrations were also measured. 2. Halothane anaesthesia decreased the turnover rate by 20%, which was similar to previously reported decreases in metabolic rates caused by natural sleep. 3. Fractional recycling of glucose carbon was little affected by halothane. 4. Comparison of values in one rat with those in another, among both conscious rats and those under halothane anaesthesia, showed that rate coefficients were inversely correlated with plasma glucose concentrations. 5. These findings indicated that halothane, in the concentration used (1.25%, v/v), had little specific effect on glucose metabolism. 6. Although equilibrium plasma glucose concentrations in different rats under halothane were widely different (4-8 mmol/l) the rates of utilization were very similar (2.5-3.1 micronmol/min per 100 g), indicating that these rates were determined by the production of glucose from gluconeogenic precursors released by basal metabolism, the rate of which is necessarily similar in different rats. 7. Among rats under halothane anaesthesia plasma insulin concentrations were negatively correlated with rate coefficients, showing that the differences between rate coefficients were mostly accounted for by differences between rats in tissue sensitivities to insulin. Thus in each 24h-starved rat, sleeping or resting, the main regulators of the plasma glucose concentrations were the rate of supply of gluconeogenic substrates from energy metabolism and the intrinsic sensitivity of the tissues to insulin. 8. We found that a commonly used deionization method of purifying glucose for determination of its specific radioactivity was inadequate.


1984 ◽  
Vol 247 (2) ◽  
pp. E166-E172 ◽  
Author(s):  
J. M. Miles ◽  
S. L. Nissen ◽  
J. E. Gerich ◽  
M. W. Haymond

Infusion of epinephrine in humans increases glucose production and decreases plasma concentrations of some essential amino acids such as leucine, while not affecting the plasma concentration of the potential gluconeogenic amino acid alanine. To determine whether epinephrine alters alanine and leucine metabolism, rates of appearance (Ra) and disappearance (Rd) of glucose, alanine, and leucine were determined in postabsorptive volunteers using [3H]glucose, [2H3]alanine, [15N]leucine, and [2H3]leucine during a 180-min infusion of epinephrine (50 ng X kg-1 X min-1). Plasma glucose (90 +/- 1 to 142 +/- 5 mg/dl) and insulin (10 +/- 1 to 16 +/- 2 micrograms/ml) increased (P less than 0.05), whereas plasma alanine concentrations did not change and plasma leucine concentrations increased (127 +/- 5 to 72 +/- 3 microM). Glucose Ra increased transiently and returned to basal values by 120 min. In contrast, alanine Ra and Rd increased identically and progressively from 5.7 +/- 0.5 to 14.5 +/- 1.9 mumol X kg-1 X min-1 by 180 min. Although leucine nitrogen Ra increased transiently and returned to basal values, leucine carbon Ra and Rd decreased (P less than 0.05) during the infusion of epinephrine. The calculated rate and percent of leucine nitrogen going to alanine increased, whereas the percent of alanine nitrogen derived from leucine remained constant. The increase in alanine Ra was entirely attributable to increased de novo synthesis because proteolysis, as estimated by leucine carbon flux, decreased.


1974 ◽  
Vol 142 (1) ◽  
pp. 171-183 ◽  
Author(s):  
Joseph Katz ◽  
Arnold Dunn ◽  
Maymie Chenoweth ◽  
Sybil Golden

1. Glucose labelled with 3H in position 2 and uniformly with 14C was administered simultaneously to rabbits and rats either as a single injection or by continuous infusion. Plasma glucose specific radioactivity and the yield of 3H in the plasma water were monitored. 2. The rates of synthesis, recycling of carbon and total body mass of glucose were calculated, without assuming a multicompartmental model and without fitting data by exponential expressions. 3. The rate of synthesis of glucose in starved-overnight rabbits was 4mg/min per kg (range 3–4.5mg/min per kg) and 25–35% of the glucose carbon was recycled. The mass of total body glucose in starved rabbits was 290mg/kg (range 220–390mg/kg). About one-third of the total body glucose equilibrates nearly instantaneously with plasma glucose. 4. In rats starved overnight, glucose synthesis was about 10mg/min per kg and recycling of carbon ranged from 30–40%. Total body mass (per kg body weight) is similar to that in rabbits. 5. The activity in plasma water after injection of [2-3H]glucose was determined. The initial rate of 3H2O formation is rapid, indicating that the major site of glucose catabolism is in the rapidly mixing pool. The curve of total body glucose radioactivity was obtained from the 3H2O yield, and total mass of glucose was calculated. This agrees with that obtained from the 3H specific-radioactivity curve.


1979 ◽  
Vol 57 (11) ◽  
pp. 1267-1274 ◽  
Author(s):  
John R. Allsop ◽  
Robert R. Wolfe ◽  
Joseph J. DiStephano III ◽  
John F. Burke

The rate of appearance of unlabelled glucose was calculated from changes in plasma glucose specific radioactivity after a single intravenous injection of labelled glucose and compared with the actual constant infusion rate of unlabelled glucose into an anaesthetized dog with all sources of endogenous glucose production surgically removed. The mean steady-state rate of appearance of unlabelled glucose calculated from the area under the specific radioactivity versus time curve was 7% higher than the actual infusion rate (n = 4), but the difference was not statistically significant. The variability in the rate calculated in this manner was, however, greater than the variability we have reported with rates determined from a primed constant infusion of tracer. Using 15- to 60- or 60- to 120-min specific radioactivity data the mean rate of appearance of glucose, calculated on the assumption of a one-pool model for glucose turnover in vivo, was approximately 60% higher than the actual infusion rate. The results also indicate that it is possible to construct multi-pool models, but it is difficult to equate specific physiological events with the individual terms of the multi-exponential equation which describes the changes in plasma glucose specific radioactivity.


1970 ◽  
Vol 48 (2) ◽  
pp. 115-122 ◽  
Author(s):  
N. Forbath ◽  
G. Hetenyi Jr.

Uniformly labelled 14C-glucose infusions were given to normal and pancreatectomized dogs. Plasma glucose and lactate specific activities were measured by appropriate methods. In normal dogs 14–28% of plasma lactate was found to originate from circulating glucose, and in diabetic dogs 11–18%. This fraction was markedly increased in normal but not significantly so in diabetic dogs during a 2-h infusion of a 6.7–11 mg/kg min glucose load. Uniformly labelled 14C-L(+)-lactate and glucose-6-3H were infused simultaneously for 5 h in normal and pancreatectomized dogs. The glucose turnover rate and incorporation of lactate carbon into glucose were calculated. Of the utilized lactate carbon, 41–49% was incorporated into plasma glucose. The recycling rate of glucose carbon via lactate was estimated to be 3–8% of the glucose turnover rate in the normal dog in the postabsorptive state. The absolute rate of gluconeogenesis from lactate was found to be increased in diabetes.


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