Oxygen consumption by bovine granulosa cells with prediction of oxygen transport in preantral follicles

2013 ◽  
Vol 25 (8) ◽  
pp. 1158 ◽  
Author(s):  
Dongxing Li ◽  
Gabe P. Redding ◽  
John E. Bronlund

The rate of oxygen consumption by granulosa cells is a key parameter in mathematical models that describe oxygen transport across ovarian follicles. This work measured the oxygen consumption rate of bovine granulosa cells in vitro to be in the range 2.1–3.3 × 10–16 mol cell–1 s–1 (0.16–0.25 mol m–3 s–1). The implications of the rates for oxygen transport in large bovine preantral follicles were examined using a mathematical model. The results indicate that oocyte oxygenation becomes increasingly constrained as preantral follicles grow, reaching hypoxic levels near the point of antrum formation. Beyond a preantral follicle radius of 134 µm, oxygen cannot reach the oocyte surface at typical values of model parameters. Since reported sizes of large bovine preantral follicles range from 58 to 145 µm in radius, this suggests that oocyte oxygenation is possible in all but the largest preantral follicles, which are on the verge of antrum formation. In preantral bovine follicles, the oxygen consumption rate of granulosa cells and fluid voidage will be the key determinants of oxygen levels across the follicle.

2021 ◽  
Vol 22 (16) ◽  
pp. 8367
Author(s):  
Hien Lau ◽  
Shiri Li ◽  
Nicole Corrales ◽  
Samuel Rodriguez ◽  
Mohammadreza Mohammadi ◽  
...  

Pre-weaned porcine islets (PPIs) represent an unlimited source for islet transplantation but are functionally immature. We previously showed that necrostatin-1 (Nec-1) immediately after islet isolation enhanced the in vitro development of PPIs. Here, we examined the impact of Nec-1 on the in vivo function of PPIs after transplantation in diabetic mice. PPIs were isolated from pancreata of 8–15-day-old, pre-weaned pigs and cultured in media alone, or supplemented with Nec-1 (100 µM) on day 0 or on day 3 of culture (n = 5 for each group). On day 7, islet recovery, viability, oxygen consumption rate, insulin content, cellular composition, insulin secretion capacity, and transplant outcomes were evaluated. While islet viability and oxygen consumption rate remained high throughout 7-day tissue culture, Nec-1 supplementation on day 3 significantly improved islet recovery, insulin content, endocrine composition, GLUT2 expression, differentiation potential, proliferation capacity of endocrine cells, and insulin secretion. Adding Nec-1 on day 3 of tissue culture enhanced the islet recovery, proportion of delta cells, beta-cell differentiation and proliferation, and stimulation index. In vivo, this leads to shorter times to normoglycemia, better glycemic control, and higher circulating insulin. Our findings identify the novel time-dependent effects of Nec-1 supplementation on porcine islet quantity and quality prior to transplantation.


1953 ◽  
Vol 9 (4) ◽  
pp. 379-390 ◽  
Author(s):  
M. REISS ◽  
EVA BRUMMEL ◽  
I. D. K. HALKERSTON ◽  
F. E. BADRICK ◽  
M. FENWICK

A technique for measuring the action of small doses of ACTH on the oxygen consumption of slices of cattle adrenal cortex is described. The oxygen consumption rate of such slices in vitro is increased by ACTH. A linear relationship between logarithm of the dose of ACTH and the percentage increase in the rate of oxygen uptake is obtained with this method, and its suitability for biological assay purposes has been investigated. The question of the specificity of this action of ACTH is discussed.


2015 ◽  
Vol 155 ◽  
pp. 35-39 ◽  
Author(s):  
I. Heredero-Bermejo ◽  
A. Criado-Fornelio ◽  
J. Soliveri ◽  
J.A. Díaz-Martín ◽  
J. Matilla-Fuentes ◽  
...  

2017 ◽  
Vol 18 (9) ◽  
pp. 1925 ◽  
Author(s):  
Karl Schoknecht ◽  
Nikolaus Berndt ◽  
Jörg Rösner ◽  
Uwe Heinemann ◽  
Jens Dreier ◽  
...  

1995 ◽  
Vol 269 (1) ◽  
pp. R226-R228 ◽  
Author(s):  
R. K. Porter ◽  
M. D. Brand

Hepatocytes were isolated from nine species of mammal of different body mass (and standard metabolic rate). The cells were incubated under identical conditions and oxygen consumption measured. The rate of oxygen consumption (per unit mass of cells) scaled with body mass with exponent -0.18. In general, there was a 5.5-fold decrease in oxygen consumption rate with a 12,500-fold increase in body mass. The decrease in oxygen consumption rate was not due to an increase in cell volume with increasing body mass but to a decrease in intrinsic metabolic activity of the cells. This novel finding confirms and explains the decrease in oxygen consumption rate measured in tissue slices from larger mammals by H. A. Krebs (Biochim. Biophys. Acta 4: 249-269, 1950) and recently by P. Couture and A. J. Hulbert [Am. J. Physiol. 268 (Regulatory Integrative Comp. Physiol. 37): R641-R650, 1995].


2021 ◽  
Vol 30 ◽  
pp. 096368972110522
Author(s):  
Hirotake Komatsu ◽  
Meirigeng Qi ◽  
Nelson Gonzalez ◽  
Mayra Salgado ◽  
Leonard Medrano ◽  
...  

Prior to transplantation into individuals with type 1 diabetes, in vitro assays are used to evaluate the quality, function and survival of isolated human islets. In addition to the assessments of these parameters in islet, they can be evaluated by multiparametric morphological scoring (0–10 points) and grading (A, B, C, D, and F) based on islet characteristics (shape, border, integrity, single cells, and diameter). However, correlation between the multiparametric assessment and transplantation outcome has not been fully elucidated. In this study, 55 human islet isolations were scored using this multiparametric assessment. The results were correlated with outcomes after transplantation into immunodeficient diabetic mice. In addition, the multiparametric assessment was compared with oxygen consumption rate of isolated islets as a potential prediction factor for successful transplantations. All islet batches were assessed and found to score: 9 points ( n = 18, Grade A), 8 points ( n = 19, Grade B), and 7 points ( n = 18, Grade B). Islets that scored 9 (Grade A), scored 8 (Grade B) and scored 7 (Grade B) were transplanted into NOD/SCID mice and reversed diabetes in 81.2%, 59.4%, and 33.3% of animals, respectively ( P < 0.0001). Islet scoring and grading correlated well with glycemic control post-transplantation ( P < 0.0001) and reversal rate of diabetes ( P < 0.05). Notably, islet scoring and grading showed stronger correlation with transplantation outcome compared to oxygen consumption rate. Taken together, a multiparametric assessment of isolated human islets was highly predictive of transplantation outcome in diabetic mice.


2022 ◽  
Vol 20 (1) ◽  
Author(s):  
Mengling Wang ◽  
Feng Zeng ◽  
Fengling Ning ◽  
Yinhang Wang ◽  
Shilin Zhou ◽  
...  

Abstract Background and aims Renal fibrosis is the common outcome in all progressive forms of chronic kidney disease. Unfortunately, the pathogenesis of renal fibrosis remains largely unexplored, among which metabolic reprogramming plays an extremely crucial role in the evolution of renal fibrosis. Ceria nanoparticles (CeNP-PEG) with strong ROS scavenging and anti-inflammatory activities have been applied for mitochondrial oxidative stress and inflammatory diseases. The present study aims to determine whether CeNP-PEG has therapeutic value for renal fibrosis. Methods The unilateral ureteral obstructive fibrosis model was used to assess the therapeutic effects in vivo. Transforming growth factor beta1-induced epithelial-to-mesenchymal transition in HK-2 cells was used as the in vitro cell model. The seahorse bioscience X96 extracellular flux analyzer was used to measure the oxygen consumption rate and extracellular acidification rate. Results In the present study, CeNP-PEG treatment significantly ameliorated renal fibrosis by increased E-cadherin protein expression, and decreased α-SMA, Vimentin and Fibronectin expression both in vitro and in vivo. Additionally, CeNP-PEG significantly reduced the ROS formation and improved the levels of mitochondrial ATP. The seahorse analyzer assay demonstrated that the extracellular acidification rate markedly decreased, whereas the oxygen consumption rate markedly increased, in the presence of CeNP-PEG. Furthermore, the mitochondrial membrane potential markedly enhanced, hexokinase 1 and hexokinase 2 expression significantly decreased after treatment with CeNP-PEG. Conclusions CeNP-PEG can block the dysregulated metabolic status and exert protective function on renal fibrosis. This may provide another therapeutic option for renal fibrosis. Graphical Abstract


2012 ◽  
Vol 24 (1) ◽  
pp. 134
Author(s):  
Y. Abe ◽  
K. Takakura ◽  
K. Kaito ◽  
T. Ogawa ◽  
M. Yokoo ◽  
...  

In the previous report, we demonstrated that bovine germinal vesicle (GV) stage oocytes vitrified using a nylon mesh holder showed an in vitro maturation rate to the metaphase II (MII) stage similar to that of fresh ones. However, cleavage and developmental rates of vitrified oocytes were low. Because mitochondria and the cytoskeleton are thought to have a central role in energy supply and cellular division in mammalian embryogenesis, it seems possible that alternation in their function in vitrified GV oocytes may contribute to subsequent lower cleavage and developmental rates. The oxygen consumption rate reflects the mitochondrial activity and its measurement may be an effective way for non-invasive evaluation of oocyte quality. In this study, to ascertain that altered mitochondrial functions and cytoskeleton may contribute to reduce the quality of oocytes after vitrification, we evaluated the distribution of active mitochondria and the cytoskeleton in vitrified oocytes. We also examined the relationship between oxygen consumption rate and the distribution of active mitochondria in vitrified oocytes. Bovine GV oocytes connected with cumulus cells were exposed to the cryoprotectant (EFS40) in a stepwise way and transferred onto a nylon mesh holder, followed by plunging them directly into liquid nitrogen. After warming, vitrified oocytes were allowed in vitro maturation. After denuding, matured oocytes were stained with a mitochondria-specific probe, rhodamine-123 and then oxygen consumption rate using an embryo respirometer (HV-403; Research Institute for Functional Peptides, Yamagata, Japan) was measured in each oocyte. According to morphological distribution of mitochondria, oocytes were classified as follows: type 1, uniform distribution; type 2, spotted distribution; and type 3, a weak fluorescence. The oxygen consumption rate of the fresh oocytes at the MII stage was significantly (P < 0.05) higher than that of vitrified oocytes (5.24 and 4 × 1015 mol–1 s–1, respectively), although there was no difference between the fresh and vitrified groups at the GV stage (5.02 and 5.06 × 1015 mol–1 s–1, respectively). The oxygen consumption rates of type 1 oocytes in fresh and vitrified groups at the MII stage tended to be higher than those of type 2 and 3 oocytes (type 1, 5.29 and 5.27; type 2, 4.99 and 4.52; type 3, 4.77 and 4.48 × 1015 mol–1 s–1, respectively). In addition, the percentage of type 1 oocytes in the fresh group was significantly (P < 0.05) higher than that in the vitrified group (59.4 and 34.3%, respectively). The matured oocytes also were stained with α-tubulin monoclonal antibody or F-phalloidin independently to examine the morphological status of microtubules or microfilament. The rates of oocytes with abnormal microtubules and microfilament in the vitrified group were 29.7 and 43.5%, respectively, showing higher rates compared with corresponding fresh oocytes (9.8%; P < 0.05 and 25.0%; P = 0.21, respectively). These results suggested that the reduction of quality and subsequent developmental competence in vitrified oocytes might be related to damages of mitochondria and cytoskeleton.


2017 ◽  
Author(s):  
Tarallo Andrea ◽  
D’Onofrio Giuseppe ◽  
Agnisola Claudio

AbstractThe three spine stickleback Gasterosteus aculeatus is a specific obligatory intermediate host for the cestode worm Schistocephalus solidus. This system is commonly used to investigate the host-parasite interaction in fishes. Despite the interesting attempts which have been made to quantify the impact of the parasite over the respiration rate of the host fish, none of the previous reports took in consideration that stickleback is diversified in different ecotypes according to its ability to made reproductive migration, from and to the sea. Here the oxygen consumption rate in specimens of three-spine stickleback collected from a non-migratory population was quantified with the aim to test if the S. solidus infection drives a change in the oxygen consumption level of the host fish. The results showed that the infected fishes have a higher rate of oxygen consumption compared with the uninfected one. The differences were due to a direct effect of the parasite, not merely to its contribution to the whole oxygen consumption rate. The data were compared with previous reports, showing that the non-migratory population was characterized by a different level of oxygen consumption rate. The differences were interpreted in terms of divergence in physiological adaptations which had to be appeared in different populations.


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