In vitro analysis of glucose metabolism and embryonic growth in postimplantation rat embryos

Development ◽  
1987 ◽  
Vol 100 (3) ◽  
pp. 431-439 ◽  
Author(s):  
S.K. Ellington
Keyword(s):  
Glucose Metabolism ◽  
Embryonic Development ◽  
Glucose Uptake ◽  
Glucose Concentration ◽  
Culture Media ◽  
Rat Embryos ◽  
Embryonic Growth ◽  
Stage Of Development

The glucose metabolism and embryonic development of rat embryos during organogenesis was studied using embryo culture. Glucose uptake and embryonic growth and differentiation of 10.5-day explants (embryos + membranes) were limited by the decreasing glucose concentration, but not the increasing concentration of metabolites, in the culture media during the second 24 h of a 48 h culture. No such limitations were found on the embryonic development of 9.5-day explants during a 48 h culture although glucose uptake was slightly reduced at very low concentrations of glucose. From the head-fold stage to the 25-somite stage of development, glucose uptake was characteristic of the stage of development of the embryo and not the time it had been in culture. Embryonic growth of 9.5-day explants was similar to that previously observed in vivo. Glucose uptake by 9.5-day explants was dependent on the surface area of the yolk sac and was independent of the glucose concentration in the culture media (within the range of 9.4 to 2.5 mM). The proportion of glucose converted to lactate was 100% during the first 42h of culture then fell to about 50% during the final 6h. The protein contents of both the extraembryonic membranes and the embryo were dependent on the glucose uptake.

scholarly journals GLUT4 in Mouse Endometrial Epithelium: Roles in Embryonic Development and Implantation

2021 ◽  
Vol 12 ◽  
Author(s):  
Yun Long ◽  
Yi-cheng Wang ◽  
Dong-zhi Yuan ◽  
Xin-hua Dai ◽  
Lin-chuan Liao ◽  
...  
Keyword(s):  
Embryonic Development ◽  
Glucose Uptake ◽  
Glucose Concentration ◽  
Endometrial Receptivity ◽  
Mouse Uterus ◽  
Uterine Fluid ◽  
Endometrial Epithelium ◽  
Interfering Rna

GLUT4 is involved in rapid glucose uptake among various kinds of cells to contribute to glucose homeostasis. Prior data have reported that aberrant glucose metabolism by GLUT4 dysfunction in the uterus could be responsible for infertility and increased miscarriage. However, the expression and precise functions of GLUT4 in the endometrium under physiological conditions remain unknown or controversial. In this study, we observed that GLUT4 exhibits a spatiotemporal expression in mouse uterus on pregnant days 1–4; its expression especially increased on pregnant day 4 during the window of implantation. We also determined that estrogen, in conjunction with progesterone, promotes the expression of GLUT4 in the endometrial epithelium in vivo or in vitro. GLUT4 is an important transporter that mediates glucose transport in endometrial epithelial cells (EECs) in vitro or in vivo. In vitro, glucose uptake decreased in mouse EECs when the cells were treated with GLUT4 small interfering RNA (siRNA). In vivo, the injection of GLUT4-siRNA into one side of the mouse uterine horns resulted in an increased glucose concentration in the uterine fluid on pregnant day 4, although it was still lower than in blood, and impaired endometrial receptivity by inhibiting pinopode formation and the expressions of leukemia inhibitory factor (LIF) and integrin ανβ3, finally affecting embryonic development and implantation. Overall, the obtained results indicate that GLUT4 in the endometrial epithelium affects embryo development by altering glucose concentration in the uterine fluid. It can also affect implantation by impairing endometrial receptivity due to dysfunction of GLUT4.

IN VITRO HYPOGLYCEMIC EFFECTS OF CAESALPINIA BONDUCELLA AND MYRISTICA FRAGRANS SEED EXTRACTS

INDIAN DRUGS ◽  
2018 ◽  
Vol 55 (02) ◽  
pp. 57-62
Author(s):  
M. A Bhutkar ◽  
◽  
S. D Bhinge ◽  
D. S. Randive ◽  
G. H Wadkar ◽  
...  
Keyword(s):  
Glucose Uptake ◽  
Plant Extracts ◽  
Glucose Concentration ◽  
Yeast Cells ◽  
In Vivo Models ◽  
Myristica Fragrans ◽  
Glucose Diffusion ◽  
Caesalpinia Bonducella

The present investigation was undertaken to assess the hypoglycemic potential of Caesalpinia bonducella (C.bonducella) and Myristica fragrans (M.fragrans), employing various in vitro techniques. The extracts of seeds of C. bonducella and M. fragrans were studied for their effects on glucose adsorption capacity, in vitro glucose diffusion, in vitro amylolysis kinetics and glucose transport across the yeast cells. It was observed that the plant extracts under study adsorbed glucose and the adsorption of glucose increased remarkably with an increase in glucose concentration. There were no significant (p≤0.05) differences between their adsorption capacities. The results of amylolysis kinetic experimental model revealed that the rate of glucose diffusion was found to be increased with time from 30 to 180 min and both the plant extracts demonstrated significant inhibitory effects on movement of glucose into external solution across dialysis membrane as compared to control. Also, the plant extracts promoted glucose uptake by the yeast cells. It was observed that the enhancement of glucose uptake was dependent on both the sample and glucose concentration. C. bonducella extract exhibited significantly higher (p≤0.05) activity than the extract of M. fragrans at all concentrations. The results of the study verified the hypoglycemic activity of the extracts of C. bonducella and M. fragrans. However, the observed effects exhibited by the extracts of seeds of C. bonducella and M. fragrans need to be confirmed by using different in vivo models and clinical trials for their effective utilization as therapeutic agents in better management of diabetes mellitus.

Maternal diabetes in vivo and high glucose concentration in vitro increases apoptosis in rat embryos

2007 ◽  
Vol 23 (1) ◽  
pp. 63-74 ◽  
Author(s):  
Mattias Gäreskog ◽  
Jonas Cederberg ◽  
Ulf J. Eriksson ◽  
Parri Wentzel
Keyword(s):  
High Glucose ◽  
Glucose Concentration ◽  
Maternal Diabetes ◽  
High Glucose Concentration ◽  
Rat Embryos

scholarly journals Bovine Oviduct Epithelial Cell-Derived Culture Media and Exosomes Improve Mitochondrial Health by Restoring Metabolic Flux during Pre-Implantation Development

2020 ◽  
Vol 21 (20) ◽  
pp. 7589
Author(s):  
Tabinda Sidrat ◽  
Abdul Aziz Khan ◽  
Myeon-Don Joo ◽  
Yiran Wei ◽  
Kyeong-Lim Lee ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Embryonic Development ◽  
Metabolic Flux ◽  
Culture Media ◽  
Tca Cycle ◽  
Fine Tuning ◽  
The Tca Cycle ◽  
Oviduct Epithelial Cell

Oviduct flushing is enriched by a wide variety of nutrients that guide the 3–4 days journey of pre-implantation embryo through the oviduct as it develops into a competent blastocyst (BL). However, little is known about the specific requirement and role of these nutrients that orchestrate the early stages of embryonic development. In this study, we aimed to characterize the effect of in vitro-derived bovine oviduct epithelial cell (BOECs) secretion that mimics the in vivo oviduct micro-fluid like environment, which allows successful embryonic development. In this study, the addition of an in vitro derived BOECs-condition media (CM) and its isolated exosomes (Exo) significantly enhances the quality and development of BL, while the hatching ability of BLs was found to be high (48.8%) in the BOECs-Exo supplemented group. Surprisingly, BOECs-Exo have a dynamic effect on modulating the embryonic metabolism by restoring the pyruvate flux into TCA-cycle. Our analysis reveals that Exo treatment significantly upregulates the pyruvate dehydrogenase (PDH) and glutamate dehydrogenase (GLUD1) expression, required for metabolic fine-tuning of the TCA-cycle in the developing embryos. Exo treatment increases the influx into TCA-cycle by strongly suppressing the PDH and GLUD1 upstream inhibitors, i.e., PDK4 and SIRT4. Improvement of TCA-cycle function was further accompanied by higher metabolic activity of mitochondria in BOECs-CM and Exo in vitro embryos. Our study uncovered, for the first time, the possible mechanism of BOECs-derived secretion in re-establishing the TCA-cycle flux by the utilization of available nutrients and highlighted the importance of pyruvate in supporting bovine in vitro embryonic development.

scholarly journals Zinc Regulates Glucose Metabolism of the Spinal Cord and Neurons and Promotes Functional Recovery after Spinal Cord Injury through the AMPK Signaling Pathway

2021 ◽  
Vol 2021 ◽  
pp. 1-27
Author(s):  
Hengshuo Hu ◽  
Nan Xia ◽  
Jiaquan Lin ◽  
Daoyong Li ◽  
Chuanjie Zhang ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Spinal Cord Injury ◽  
Nervous System ◽  
Glucose Metabolism ◽  
Glucose Uptake ◽  
Functional Recovery ◽  
Glucose Metabolism Disorders ◽  
Cord Injury

Spinal cord injury (SCI) is a traumatic disease that can cause severe nervous system dysfunction. SCI often causes spinal cord mitochondrial dysfunction and produces glucose metabolism disorders, which affect neuronal survival. Zinc is an essential trace element in the human body and plays multiple roles in the nervous system. This experiment is intended to evaluate whether zinc can regulate the spinal cord and neuronal glucose metabolism and promote motor functional recovery after SCI. Then we explore its molecular mechanism. We evaluated the function of zinc from the aspects of glucose uptake and the protection of the mitochondria in vivo and in vitro. The results showed that zinc elevated the expression level of GLUT4 and promoted glucose uptake. Zinc enhanced the expression of proteins such as PGC-1α and NRF2, reduced oxidative stress, and promoted mitochondrial production. In addition, zinc decreased neuronal apoptosis and promoted the recovery of motor function in SCI mice. After administration of AMPK inhibitor, the therapeutic effect of zinc was reversed. Therefore, we concluded that zinc regulated the glucose metabolism of the spinal cord and neurons and promoted functional recovery after SCI through the AMPK pathway, which is expected to become a potential treatment strategy for SCI.

Insulin binding and action in adipocytes in vitro in relation to insulin action in vivo in young and middle-aged subjects

1986 ◽  
Vol 113 (1) ◽  
pp. 88-92 ◽  
Author(s):  
Hannele Yki-Jarvinen ◽  
Tuula Kiviluoto ◽  
Esko A. Nikkila
Keyword(s):  
Adipose Tissue ◽  
Glucose Metabolism ◽  
Glucose Uptake ◽  
Glucose Transport ◽  
Young Group ◽  
Middle Aged ◽  
Young Subjects ◽  
Aged Subjects

Abstract. The effect of age on glucose metabolism in vivo was compared to that found in adipocytes in vitro in young (n = 8, age 23 to 31 years) and middle-aged (n = 7, age 37 to 55 years) non-diabetic subjects. During the OGTT, the incremental glucose or insulin areas did not differ significantly between the groups. Fasting and 2 h plasma glucose (P < 0.01) and the 2 h plasma insulin (P < 0.05) levels were, however, slightly higher in the middle-aged than in the young group. During iv induced hyperinsulinaemia (~ 85 mU/l), rates of glucose uptake were comparable between the middle-aged (6.32 ± 0.94 mg/kg/min) and the young subjects (7.56 ± 0.78 mg/kg/min, P > 0.5). In fat cells, both basal and insulin stimulated rates of glucose transport were 35% lower (P < 0.05) in the middle-aged than in the young subjects. Basal and insulin stimulated rates of glucose oxidation and lipogenesis were both markedly lower (P <0.01) in the middle-aged than in the young group. The rates of glucose transport, oxidation and lipogenesis were inversely related to age, whereas none of these parameters was related to fat cell size. In conclusion, adipocyte glucose metabolism in middle-aged healthy subjects was markedly impaired. In contrast, rates of glucose uptake during iv hyperinsulinaemia and glucose responses during hyperinsulinaemia in the OGTT were comparable in young and middle-aged subjects. These results indicate first, that changes in adipose tissue glucose metabolism by aging do not parallel changes in whole body glucose metabolism and thus are specific for adipose tissue. Secondly, insulin-mediated glucose uptake is not reduced in middle-aged subjects. Thus, the small elevation in fasting and 2-h glucose levels may reflect a reduction in glucose uptake by non-insulin dependent pathways by aging.

Glucose Metabolism Characterization During Mouse In Vitro Maturation Identifies Alterations In Cumulus Cells†

2021 ◽  
Author(s):  
Nazli Akin ◽  
Lucia von Mengden ◽  
Anamaria-Cristina Herta ◽  
Katy Billooye ◽  
Julia Leersum ◽  
...  
Keyword(s):  
Glucose Metabolism ◽  
In Vitro Maturation ◽  
Culture Media ◽  
Cumulus Cells ◽  
Routine Practice ◽  
Glycolytic Activity ◽  
Direct Measurements ◽  
Final Maturation

Abstract In vitro maturation (IVM) is an assisted reproduction technique with reduced hormone-related side effects. Several attempts to implement IVM in routine practice have failed, primarily due to its relatively low efficiency compared to conventional in vitro fertilization (IVF). Recently, capacitation (CAPA)-IVM, a novel two-step IVM method, has improved the embryology outcomes through synchronizing the oocyte nuclear and cytoplasmic maturation. However, the efficiency gap between CAPA-IVM and conventional IVF is still noticeable especially in the numerical production of good quality embryos. Considering the importance of glucose for oocyte competence, its metabolization is studied within both in vivo and CAPA-IVM matured mouse cumulus-oocyte-complexes (COCs) through direct measurements in both cellular compartments, from transcriptional and translational perspectives, to reveal metabolic shortcomings within the CAPA-IVM COCs. These results confirmed that within in vivo COC, cumulus cells are highly glycolytic, whereas oocytes, with low glycolytic activity, are deviating their glucose towards pentose phosphate pathway. No significant differences were observed in the CAPA-IVM oocytes compared to their in vivo counterparts. However, their cumulus cells exhibited a precocious increase of glycolytic activity during the pre-maturation culture step and activity was decreased during the IVM step. Here, specific alterations in mouse COC glucose metabolism due to CAPA-IVM culture were characterized using direct measurements for the first time. Present data show that, while CAPA-IVM cumulus cells are able to utilize glucose, their ability to support oocytes during final maturation is impaired. Future CAPA-IVM optimization strategies could focus on adjusting culture media energy substrate concentrations and/or implementing co-culture strategies.

Action of epinephrine on muscle glucose uptake depending on Ca++ and phosphate

1965 ◽  
Vol 209 (2) ◽  
pp. 359-364 ◽  
Author(s):  
Michio Ui
Keyword(s):  
Glucose Metabolism ◽  
Glucose Uptake ◽  
Inorganic Phosphate ◽  
Glucose Utilization ◽  
Rat Diaphragm ◽  
Muscle Tissues ◽  
Ph Level ◽  
Diaphragm In Vitro

Studies have been made of the involvement of inorganic phosphate (Pi) and Ca++ in the mechanism by which epinephrine-induced inhibition of muscle glucose utilization was abolished during either alkalosis in vivo or incubation of the isolated rat diaphragm in vitro at a higher pH level. An increase in the concentration of Pi in muscle tissues was closely associated with prevention of the inhibitory action of epinephrine on glucose uptake. The interrelationship of Ca++ and Pi in aqueous solutions, and the additional observations that glucose uptake by rat diaphragm was accelerated in anaerobiosis only in the absence of Ca++, indicate a significance of Ca++ in muscle glucose metabolism. Assay of hexokinase activity in cell-free muscle preparations revealed that the inhibition of the enzyme activity by glucose 6-phosphate was profoundly influenced by the presence of Ca++ and Pi and was dependent on the concentration of ATP. It is suggested that Ca++ may provide the primary point of influence of epinephrine on glucose metabolism of the muscle.

scholarly journals 135 REGULATION OF GLUCOSE METABOLISM TO DECREASE LIPID CONTENT OF IN VIRTO-PRODUCED BOVINE EMBRYOS

2005 ◽  
Vol 17 (2) ◽  
pp. 218 ◽  
Author(s):  
J. De La Torre-Sanchez ◽  
D. Gardner ◽  
K. Preis ◽  
G. Seidel Jr
Keyword(s):  
Glucose Metabolism ◽  
Glucose Uptake ◽  
Surface Area ◽  
Lipid Accumulation ◽  
Lactate Production ◽  
Pentose Phosphate ◽  
Developmental Competence ◽  
Metabolic Regulators

Our objective was to improve normality of embryos produced in vitro with regulators of carbohydrate metabolism at doses optimized in earlier experiments. Eight- to 16-cell embryos were produced in vitro in the G1/G2 system (chemically defined sequential medium with recombinant human serum albumin), and then cultured 3 days in G2 containing metabolic regulators as follows: phenazine ethosulfate (PES), 0.3 μM; NaN3, 27 μM; 2,4-dinitrophenol (DNP), 30 μM; and control. The following responses were analyzed by ANOVA in 2 to 4 replicates of 8–12 embryos each: glucose uptake and metabolism (uptake measured by microfluorometry of medium after incubating an embryo 3 h; metabolism measured as 3H2O released after incubating an embryo 3 h in medium containing 5-3H glucose), % of glucose metabolized via the pentose phosphate pathway (PPP rate), lactate production, glycolysis (% of lactate produced from glucose taken up on a molar basis), lipid accumulation (number of >2 μM Sudan Black B positive granules/103 μm2), % live Day 14 embryos recovered from embryos transferred to recipients at Day 7, and average surface area of embryos collected. In vivo-derived embryos were included as a second control for lipid evaluation. PES-treated embryos had higher glucose metabolism (P < 0.05) and lower glucose uptake (P < 0.01) than embryos in NaN3 and tended to have a higher PPP rate (P < 0.11) than controls; however, glycolysis was higher for PES than other treatments (P < 0.01) (Table 1). Lipid accumulation of embryos from PES was markedly lower than any other in vitro treatments (P < 0.01), but higher than in vivo embryos (3.31 ± 2.78 lipid granules) (P < 0.01). NaN3- and DNP-treated embryos both accumulated lipid similar to in vitro controls. No treatment differences were found in developmental competence when Day 7 embryos were transferred to recipients and recovered 1 week later (43 to 54% live embryos recovered), nor were there any significant differences (P > 0.1) in surface area. Embryos exposed to PES at the compaction and post-compaction stages accumulated much less lipid than controls or embryos exposed to other metabolic regulators, making this a very promising treatment. PES oxidizes NADPH; the molecular mechanism of PES appears to involve increased flux of glucose through the PPP while decreasing availability of NADPH for fatty acid synthesis. Table 1. Response of embryos to metabolic regulators

scholarly journals Protein kinase N2 regulates AMP kinase signaling and insulin responsiveness of glucose metabolism in skeletal muscle

2017 ◽  
Vol 313 (4) ◽  
pp. E483-E491 ◽  
Author(s):  
Maxwell A. Ruby ◽  
Isabelle Riedl ◽  
Julie Massart ◽  
Marcus Åhlin ◽  
Juleen R. Zierath
Keyword(s):  
Skeletal Muscle ◽  
Protein Kinase ◽  
Glucose Metabolism ◽  
Glucose Uptake ◽  
Muscle Metabolism ◽  
Whole Body ◽  
Skeletal Muscle Metabolism ◽  
Ampk Signaling

Insulin resistance is central to the development of type 2 diabetes and related metabolic disorders. Because skeletal muscle is responsible for the majority of whole body insulin-stimulated glucose uptake, regulation of glucose metabolism in this tissue is of particular importance. Although Rho GTPases and many of their affecters influence skeletal muscle metabolism, there is a paucity of information on the protein kinase N (PKN) family of serine/threonine protein kinases. We investigated the impact of PKN2 on insulin signaling and glucose metabolism in primary human skeletal muscle cells in vitro and mouse tibialis anterior muscle in vivo. PKN2 knockdown in vitro decreased insulin-stimulated glucose uptake, incorporation into glycogen, and oxidation. PKN2 siRNA increased 5′-adenosine monophosphate-activated protein kinase (AMPK) signaling while stimulating fatty acid oxidation and incorporation into triglycerides and decreasing protein synthesis. At the transcriptional level, PKN2 knockdown increased expression of PGC-1α and SREBP-1c and their target genes. In mature skeletal muscle, in vivo PKN2 knockdown decreased glucose uptake and increased AMPK phosphorylation. Thus, PKN2 alters key signaling pathways and transcriptional networks to regulate glucose and lipid metabolism. Identification of PKN2 as a novel regulator of insulin and AMPK signaling may provide an avenue for manipulation of skeletal muscle metabolism.

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