Enzymatic fluorometric microplate assay for quantitative analysis of 3-hydroxybutyric acid in mouse plasma

Takayama, Naoyuki; Ohara, Tatsuya; Yamamoto, Norio

doi:10.1016/j.ab.2012.03.011

Cited by 9 publications

(4 citation statements)

References 9 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The results found a preliminary relationship between obesity and metabolites such as hexanoic acid [ 65 ]. 3-Hydroxybutyric acid (3HB) is a ketone body and acts as an indicator of energy balance and a central regulator of energy homeostasis [ 66 ]. Studies have shown that the peroxisome proliferator-activated receptor alpha (PPARα) -dependent activation and promotion of fatty acid utilization in the liver induces the production of 3-HB [ 67 ], but the relevant mechanism deeply related to obesity-T2DM is not clear yet.…”

Section: Discussionmentioning

confidence: 99%

Fecal microbiota transplantation alters the susceptibility of obese rats to type 2 diabetes mellitus

Zhang

Zhou

Zhan

et al. 2020

Aging

View full text Add to dashboard Cite

Obesity is one of the susceptibility factors for type 2 diabetes (T2DM), both of which could accelerate the aging of the body and bring many hazards. A causal relationship is present between intestinal microbiota and body metabolism, but how the microbiota play a role in the progression of obesity to T2DM has not been elucidated. In this study, we transplanted healthy or obese-T2DM intestinal microbiota to ZDF and LZ rats, and used 16S rRNA and targeted metabonomics to evaluate the directional effect of the microbiota on the susceptibility of obese rats to T2DM. The glycolipid metabolism phenotype could be changed bidirectionally in obese rats instead of in lean ones. One possible mechanism is that the microbiota and metabolites alter the structure of the intestinal tract, and improve insulin and leptin resistance through JAK2 / IRS / Akt pathway. It is worth noting that 7 genera, such as Lactobacillus , Clostridium and Roche, can regulate 15 metabolites, such as 3-indolpropionic acid, acetic acid and docosahexaenoic acid, and have a significant improvement on glycolipid metabolism phenotype. Attention to intestinal homeostasis may be the key to controlling obesity and preventing T2DM.

show abstract

Section: Discussionmentioning

confidence: 99%

Fecal microbiota transplantation alters the susceptibility of obese rats to type 2 diabetes mellitus

Zhang

Zhou

Zhan

et al. 2020

Aging

View full text Add to dashboard Cite

show abstract

“…Thus, resazurin is a useful chemical tool for quantifying cellular levels of NAD(P)H or diaphorase. Indeed, the resazurin-diaphorase system may be employed for defining any biochemical reaction that generates NAD(P)H (Takayama et al, 2012).…”

Section: Commentary Background Informationmentioning

confidence: 99%

Measurement of Glucose Uptake in Cultured Cells

Yamamoto¹,

Ueda-Wakagi

Sato³

et al. 2015

CP Pharmacology

Self Cite

120

View full text Add to dashboard Cite

Facilitative glucose uptake transport systems are ubiquitous in animal cells and are responsible for transporting glucose across cell surface membranes. Evaluation of glucose uptake is crucial in the study of numerous diseases and metabolic disorders such as myocardial ischemia, diabetes mellitus, and cancer. Detailed in this unit are laboratory methods for assessing glucose uptake into mammalian cells. The unit is divided into five sections: (1) a brief overview of glucose uptake assays in cultured cells; (2) a method for measuring glucose uptake using radiolabeled 3-O-methylglucose; (3) a method for measuring glucose uptake using radiolabeled 2-deoxyglucose (2DG); (4) a microplate method for measuring 2DG-uptake using an enzymatic, fluorometric assay; and (5) a microplate-based method using a fluorescent analog of 2DG.

show abstract

“…Classical enzymatic-based metabolite quantitation, on the other hand, provides a cost-effective, readily accessible, sensitive means for quantitative analysis of metabolites [26]. Although unable to provide simultaneous information on the quantities of multiple metabolites, enzymatic quantitation offers the ability to simultaneously quantitate a given metabolite across many samples on a high-throughput platform [27]–[29].…”

Section: Introductionmentioning

confidence: 99%

“…One strategy involves the coupling of NAD(P)H synthesis to mitochondrial lipoyl dehydrogenase (diaphorase) which activates chemical substrates for chromogenic or fluorescence assays. Resazurin is one such compound that can be used for enzymatically-coupled and cell viability assays [27], [33], [34]. Here, we report a novel enzymatic assay for the quantitation of GABA and successfully use this assay to characterize changes in cellular GABA in PNEC cells under metabolic stress.…”

Section: Introductionmentioning

confidence: 99%

A Fluorescence-Coupled Assay for Gamma Aminobutyric Acid (GABA) Reveals Metabolic Stress-Induced Modulation of GABA Content in Neuroendocrine Cancer

Ippolito

Piwnica‐Worms

2014

PLoS ONE

View full text Add to dashboard Cite

Pathways involved in the synthesis of the neurotransmitter gamma-aminobutyric acid (GABA) have been implicated in the pathogenesis of high grade neuroendocrine (NE) neoplasms as well as neoplasms from a non-NE lineage. Using The Cancer Genome Atlas, overexpression of the GABA synthetic enzyme, glutamate decarboxylase 1 (GAD1), was found to be associated with decreased disease free-survival in prostate adenocarcinoma and decreased overall survival in clear cell renal cell carcinomas. Furthermore, GAD1 was found to be expressed in castrate-resistant prostate cancer cell lines, but not androgen-responsive cell lines. Using a novel fluorescence-coupled enzymatic microplate assay for GABA mediated through reduction of resazurin in a prostate neuroendocrine carcinoma (PNEC) cell line, acid microenvironment-induced stress increased GABA levels while alkaline microenvironment-induced stress decreased GABA through modulation of GAD1 and glutamine synthetase (GLUL) activities. Moreover, glutamine but not glucose deprivation decreased GABA through modulation of GLUL. Consistent with evidence in prokaryotic and eukaryotic organisms that GABA synthesis mediated through GAD1 may play a crucial role in surviving stress, GABA may be an important mediator of stress survival in neoplasms. These findings identify GABA synthesis and metabolism as a potentially important pathway for regulating cancer cell stress response as well as a potential target for therapeutic strategies.

show abstract

Enzymatic fluorometric microplate assay for quantitative analysis of 3-hydroxybutyric acid in mouse plasma

Cited by 9 publications

References 9 publications

Fecal microbiota transplantation alters the susceptibility of obese rats to type 2 diabetes mellitus

Fecal microbiota transplantation alters the susceptibility of obese rats to type 2 diabetes mellitus

Measurement of Glucose Uptake in Cultured Cells

A Fluorescence-Coupled Assay for Gamma Aminobutyric Acid (GABA) Reveals Metabolic Stress-Induced Modulation of GABA Content in Neuroendocrine Cancer

Contact Info

Product

Resources

About