scholarly journals 287 INHIBITION OF THE PENTOSE PHOSPHATE PATHWAY RESULTS IN MEIOTIC ARREST IN PORCINE OOCYTES THAT CAN BE OVERCOME BY THE ADDITION OF PATHWAY COFACTORS AND END PRODUCTS

2005 ◽  
Vol 17 (2) ◽  
pp. 293 ◽  
Author(s):  
R. Krisher

Glucose metabolism is an indicator of oocyte developmental competence, and is also correlated with meiotic maturation. In vitro maturation of porcine oocytes with the pentose phosphate pathway (PPP) inhibitor diphenyleneiodonium (DPI) blocks meiotic progression to metaphase II. The objectives of this study were (1) to examine the reversibility of meiotic arrest induced by DPI; and (2) to overcome metabolically induced meiotic arrest by the addition of PPP end products and cofactors downstream of DPI inhibition. Oocytes were matured for 40 h in standard (defined) maturation media containing 0, 25, 50, or 100 nM DPI. At that time half the oocytes in each treatment (TRT) were fixed, and half were moved into standard maturation medium with no DPI for an additional 40 h, at which time all remaining oocytes were fixed. Two oocytes were matured for 40 h in one of 11 media: standard (defined) maturation medium (STND), standard with 50 nM DPI (DPI), standard with 50 nM DPI and 0.25, 2.5, or 5 mM phosphoribose diphosphate (PRPP), nicotinamide adenine dinucleotide phosphate (NADP), or ribose-5-phosphate (R5P). Additionally, 10 mM R5P and 12.5 mM PRPP were examined. All oocytes were fixed. Oocytes were assigned a meiotic score; germinal vesicle (GV) = 1, GV breakdown (GVBD) = 2, condensed chromatin (CC) = 3, metaphase I (MI) = 4, anaphase (A) = 5, telophase (T) = 6, and metaphase II (MII) = 7. Immature oocytes were classified as those at GV or GVBD stages, and mature oocytes as those at A, T or MII stages. Data were analyzed by ANOVA and are presented as mean ± SEM. After 40 h of arrest (n = 79–87/TRT), increasing concentrations of DPI significantly increased the % of immature oocytes (0, 7.2 ± 2.9; 25, 26.4 ± 4.8; 50, 53.2 ± 5.7; 100, 75.9 ± 4.8) and decreased the % of mature oocytes (0, 73.5 ± 4.9; 25, 52.9 ± 5.4; 50, 20.3 ± 4.6; 100, 0). After an additional 40 hours in standard maturation medium (n = 89–93/TRT), there was no difference in the % of immature oocytes between treatments (0, 7.5 ± 2.8; 25, 14.4 ± 3.7; 50, 13.0 ± 3.5; 100, 9.0 ± 3.0) although the % of mature oocytes significantly decreased with increasing DPI concentration (0, 90.3 ± 3.1; 25, 68.9 ± 4.9; 50, 35.9 ± 5.0; 100, 10.1 ± 3.2). Data from experiment 2 are presented below. Meiotic maturation is significantly inhibited by DPI in a dose-dependent manner. Ability of the oocyte to reach MII following 40 h of arrest is also concentration-dependent, although all treatments resulted in GVBD following removal from DPI. Metabolic arrest can be overcome, resulting in numbers of mature oocytes equal to standard controls, by NADP and PRPP but only moderately by R5P. These data demonstrate that glucose metabolism via the PPP is a critical control mechanism of meiotic maturation in porcine oocytes. Table 1. Effect of PPP cofactors and end products on overcoming metabolically induced meiotic arrest in porcine oocytes

2014 ◽  
Vol 26 (7) ◽  
pp. 931 ◽  
Author(s):  
Cynthia Gutnisky ◽  
Gabriel C. Dalvit ◽  
Jeremy G. Thompson ◽  
Pablo D. Cetica

The relationship between pentose phosphate pathway (PPP) activity in cumulus–oocyte complexes (COCs) and oxidative and mitochondrial activity in bovine oocytes was evaluated with the aim of analysing the impact of two inhibitors (NADPH and 6-aminonicotinamide (6-AN)) and a stimulator (NADP) of the key enzymes of the PPP on the maturation rate, oxidative and mitochondrial activity and the mitochondrial distribution in oocytes. The proportion of COCs with measurable PPP activity (assessed using brilliant cresyl blue staining), glucose uptake, lactate production and meiotic maturation rate diminished when 6-AN (0.1, 1, 5 and 10 mM for 22 h) was added to the maturation medium (P < 0.05). The addition of NADPH did not modify glucose uptake or lactate production, but reduced PPP activity in COCs and meiotic maturation rates (P < 0.05). The presence of NADP (0.0125, 0.125, 1.25 and 12.5 mM for 22 h of culture) in the maturation medium had no effect on PPP activity in COCs, glucose uptake, lactate production and meiotic maturation rate. However, in the absence of gonadotropin supplementation, NADP stimulated both glucose uptake and lactate production at 12.5 mM (the highest concentration tested; P < 0.05). NADP did not modify cleavage rate, but decreased blastocyst production (P < 0.05). During IVM, oocyte oxidative and mitochondrial activity was observed to increase at 15 and 22 h maturation, which was also related to progressive mitochondrial migration. Inhibiting the PPP with 6-AN or NADPH led to reduced oxidative and mitochondrial activity compared with the respective control groups and inhibition of mitochondrial migration (P < 0.05). Stimulation of the PPP with NADP increased oxidative and mitochondrial activity at 9 h maturation (P < 0.05) and delayed mitochondrial migration. The present study shows the significance of altering PPP activity during bovine oocyte IVM, revealing that there is a link between the activity of the PPP and the oxidative status of the oocyte.


FEBS Letters ◽  
2003 ◽  
Vol 555 (2) ◽  
pp. 307-310 ◽  
Author(s):  
Rolando B. Ceddia ◽  
George J. Bikopoulos ◽  
Arthur J. Hilliker ◽  
Gary Sweeney

2014 ◽  
Vol 34 (4) ◽  
pp. 724-734 ◽  
Author(s):  
Eva MF Brekke ◽  
Tora S Morken ◽  
Marius Widerøe ◽  
Asta K Håberg ◽  
Ann-Mari Brubakk ◽  
...  

The neonatal brain is vulnerable to oxidative stress, and the pentose phosphate pathway (PPP) may be of particular importance to limit the injury. Furthermore, in the neonatal brain, neurons depend on de novo synthesis of neurotransmitters via pyruvate carboxylase (PC) in astrocytes to increase neurotransmitter pools. In the adult brain, PPP activity increases in response to various injuries while pyruvate carboxylation is reduced after ischemia. However, little is known about the response of these pathways after neonatal hypoxia-ischemia (HI). To this end, 7-day-old rats were subjected to unilateral carotid artery ligation followed by hypoxia. Animals were injected with [1,2-13C]glucose during the recovery phase and extracts of cerebral hemispheres ipsi- and contralateral to the operation were analyzed using 1H- and 13C-NMR (nuclear magnetic resonance) spectroscopy and high-performance liquid chromatography (HPLC). After HI, glucose levels were increased and there was evidence of mitochondrial hypometabolism in both hemispheres. Moreover, metabolism via PPP was reduced bilaterally. Ipsilateral glucose metabolism via PC was reduced, but PC activity was relatively preserved compared with glucose metabolism via pyruvate dehydrogenase. The observed reduction in PPP activity after HI may contribute to the increased susceptibility of the neonatal brain to oxidative stress.


2016 ◽  
Vol 2016 ◽  
pp. 1-13 ◽  
Author(s):  
Pedro Cisternas ◽  
Paulina Salazar ◽  
Carmen Silva-Álvarez ◽  
L. Felipe Barros ◽  
Nibaldo C. Inestrosa

In the last few years, several reports have proposed that Wnt signaling is a general metabolic regulator, suggesting a role for this pathway in the control of metabolic flux. Wnt signaling is critical for several neuronal functions, but little is known about the correlation between this pathway and energy metabolism. The brain has a high demand for glucose, which is mainly used for energy production. Neurons use energy for highly specific processes that require a high energy level, such as maintaining the electrical potential and synthesizing neurotransmitters. Moreover, an important metabolic impairment has been described in all neurodegenerative disorders. Despite the key role of glucose metabolism in the brain, little is known about the cellular pathways involved in regulating this process. We report here that Wnt5a induces an increase in glucose uptake and glycolytic rate and an increase in the activity of the pentose phosphate pathway; the effects of Wnt5a require the intracellular generation of nitric oxide. Our data suggest that Wnt signaling stimulates neuronal glucose metabolism, an effect that could be important for the reported neuroprotective role of Wnt signaling in neurodegenerative disorders.


1976 ◽  
Vol 22 (9) ◽  
pp. 1374-1380 ◽  
Author(s):  
N. Jayanthi Bai ◽  
M. Ramachandra Pai ◽  
P. Suryanarayana Murthy ◽  
T. A. Venkitasubramanian

Glucose metabolism in Mycobacterium smegmatis was investigated by the radiorespirometric method and by assaying for key enzymes of the major energy-yielding pathways. Glucose is oxidized in this organism mainly through the Embden–Meyerhof–Parnas pathway, irrespective of the carbon source used for growth. The pentose phosphate pathway plays only a minor role and its extent depends on the carbon source used for growth. Enzymes of glycolytic and oxidative pathways were detected in cells grown on glucose, glycerol, or pyruvate but enzymes of the Entner–Doudoroff pathway could be detected only in glucose-grown cells. Labeled acetate is utilized by cells cultured on glucose, glycerol, and pyruvate. In all cases more of C1 of acetate was converted to CO2 while incorporation into cellular constituents was maximum from C2 of acetate.


Nature ◽  
1964 ◽  
Vol 201 (4921) ◽  
pp. 825-827 ◽  
Author(s):  
PETER BEACONSFIELD ◽  
AMILCARE CARPI

2006 ◽  
Vol 18 (2) ◽  
pp. 279 ◽  
Author(s):  
L. Tubman ◽  
A. Peter ◽  
R. Krisher

Diphenyleneiodonium (DPI), an inhibitor of the pentose phosphate pathway (PPP), arrests nuclear maturation of porcine oocytes. This inhibition is reversed using products or cofactors of PPP such as nicotinamide adenine dinucleotide phosphate (NADP), phosphoribose diphosphate (PRPP), and ribose-5-phosphate (R5P). The objective of this study was to determine the relationship between DPI-mediated meiotic inhibition, reversal of this inhibition, and metabolism of in vitro-matured (IVM) porcine oocytes. Oocytes were aspirated, searched, and selected in the presence of DPI, with the exception of control oocytes. Oocytes were then matured in one of five treatments for 40 h in 7% CO2 in air at 39°C in defined Purdue Porcine Medium for maturation (PPMmat). Treatments included control, 50 nM DPI (DPI), DPI + 5 mM NADP (NADP), DPI + 12.5 mM PRPP (PRPP), and DPI + 10 mM R5P (R5P). Following IVM, oocytes were denuded by vortexing. Glycolysis and PPP activities were measured in 4 μL hanging drops containing labeled glucose (0.0125 mM 5-3H glucose and 0.482 mM 1-14C glucose, respectively) for 3 h in 6% CO2. Oocytes were then individually fixed in a 3:2:1 solution of ethanol:acetic acid:chloroform and stained with aceto-orcein for determination of meiotic stage (germinal vesicle = 1 through metaphase II = 7). Data were analyzed using one-way ANOVA. The use of DPI inhibited PPP and nuclear maturation; additionally glycolysis was decreased by DPI compared to control. Addition of NADP and PRPP increased both metabolic pathways and nuclear maturation compared to DPI. R5P restored glycolysis and nuclear maturation to control levels, and PPP to above the control level. There were no significant differences among meiotic stages relative to glycolytic activity. PPP activity was significantly different (values with different superscripts; P < 0.05) among oocytes of different meiotic stages (germinal vesicle = 0.24 ± 0.03ad, germinal vesicle breakdown = 0.40 ± 0.05bcde, condensed chromatin = 0.44 ± 0.05bcd, metaphase I = 0.45 ± 0.12abcd, anaphase = 0.76 ± 0.50abcde, telophase = 0.92 ± 0.17be, metaphase II = 0.74 ± 0.08be). Percentages of oocytes reaching MII were 43.48 (control), 2.08 (DPI), 28.30 (NADP), 18.18 (PRPP), and 46.94 (R5P). These results demonstrate that the PPP is a critical control mechanism for nuclear maturation of porcine oocytes, as inhibition of this metabolic pathway resulted in arrest of nuclear maturation. Addition of PPP cofactors or end products to the arresting medium led to reversal of inhibition as demonstrated by restoration of PPP activity resulting in nuclear maturation. Table 1. Meiotic stage, glycolysis, and pentose phosphate pathway activity after in vitro maturation of porcine oocytes


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