Enhanced detection of hydrogen sulfide generated in cell culture using an agar trap method

Kartha, Reena V.; Zhou, Jie; Hovde, Laurie B.; Cheung, Belinda W.Y.; Schröder, H.

doi:10.1016/j.ab.2012.01.001

Cited by 37 publications

(31 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To examine specific microbial metabolites that have known impact on host physiology, we determined relative short chain fatty acid (SCFA) concentration in cecal contents by GC-MS-MS (Renom et al, 2001) (Figure 3D–F) as well as proxy H 2 S production via methylene blue culture assay (Kartha et al, 2012). Since many of the oscillatory OTUs under RC-fed conditions belonged to the family Lachnospiraceae, which is well characterized for its capacity to synthesize butyrate under specific dietary conditions (Vital et al, 2014), we hypothesized that diurnal variation of SCFAs would be observed in RC-fed mice.…”

Section: Resultsmentioning

confidence: 99%

“…Fresh samples were collected, weighed, homogenized in water, and centrifuged at 13,000 x g. Supernatants were acidified and Isobutyric acid (3mM) was added (internal standard). SCFAs were extracted using diethyl ether, derivatized using MTBS-TFA, and run on a Varian Saturn 2000 GC-MS-MS. H 2 S production potential of cecal and fecal microbes was determined via the Agar-trap method (Kartha et al, 2012). Fresh cecal or fecal contents were collected, weighed, transferred into an anaerobic chamber, and resuspended in Bru media based on weight.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Effects of Diurnal Variation of Gut Microbes and High-Fat Feeding on Host Circadian Clock Function and Metabolism

Leone

Gibbons

Martinez

et al. 2015

Cell Host & Microbe

709

767

View full text Add to dashboard Cite

SUMMARY Circadian clocks and metabolism are inextricably intertwined, where central and hepatic circadian clocks coordinate metabolic events in response to light-dark and sleep-wake cycles. We reveal an additional key element involved in maintaining host circadian rhythms, the gut microbiome. Despite persistence of light-dark signals, germ-free mice fed low or high fat diets exhibit markedly impaired central and hepatic circadian clock gene expression and do not gain weight compared to conventionally-raised counterparts. Examination of gut microbiota in conventionally-raised mice showed differential diurnal variation in microbial structure and function dependent upon dietary composition. Additionally, specific microbial metabolites induced under low or high fat feeding, particularly short chain fatty acids, but not hydrogen sulfide, directly modulate circadian clock gene expression within hepatocytes. These results underscore the ability of microbially-derived metabolites to regulate or modify central and hepatic circadian rhythm and host metabolic function, the latter following intake of a Westernized diet.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Effects of Diurnal Variation of Gut Microbes and High-Fat Feeding on Host Circadian Clock Function and Metabolism

Leone

Gibbons

Martinez

et al. 2015

Cell Host & Microbe

709

767

View full text Add to dashboard Cite

show abstract

“…During this treatment period, cells were incubated at 37C in sealed multiwell plates with gas traps containing sterile agar containing alkaline zinc acetate (Supplementary Material). Following incubation, the agar was assayed for H 2 S content by spectrometry, as described previously (7). H 2 S content was reported relative to the untreated cells.…”

mentioning

confidence: 99%

“…As proof of principle, we chose to measure cellular H 2 S production using solid agar (6,7). Agar containing alkaline zinc acetate was applied to the gas trap.…”

mentioning

confidence: 99%

See 1 more Smart Citation

A Gas Trapping Method for High-Throughput Metabolic Experiments

et al. 2018

View full text Add to dashboard Cite

Research into cellular metabolism has become more high-throughput, with typical cell-culture experiments being performed in multiwell plates (microplates). This format presents a challenge when trying to collect gaseous products, such as carbon dioxide (CO2), which requires a sealed environment and a vessel separate from the biological sample. To address this limitation, we developed a gas trapping protocol using perforated plastic lids in sealed cell-culture multiwell plates. We used this trap design to measure CO2 production from glucose and fatty acid metabolism, as well as hydrogen sulfide production from cysteine-treated cells. Our data clearly show that this gas trap can be applied to liquid and solid gas-collection media and can be used to study gaseous product generation by both adherent cells and cells in suspension. Since our gas traps can be adapted to multiwell plates of various sizes, they present a convenient, cost-effective solution that can accommodate the trend toward high-throughput measurements in metabolic research.

show abstract

Hydrogen Sulfide Actions in the Vasculature

Mendiola

Naik

Bosc

et al. 2021

Comprehensive Physiology

View full text Add to dashboard Cite

Hydrogen sulfide (H 2 S) is a small, gaseous molecule with poor solubility in water that is generated by multiple pathways in many species including humans. It acts as a signaling molecule in many tissues with both beneficial and pathological effects. This article discusses its many actions in the vascular system and the growing evidence of its role to regulate vascular tone, angiogenesis, endothelial barrier function, redox, and inflammation. Alterations in some disease states are also discussed including potential roles in promoting tumor growth and contributions to the development of metabolic disease.

show abstract

Enhanced detection of hydrogen sulfide generated in cell culture using an agar trap method

Cited by 37 publications

References 32 publications

Effects of Diurnal Variation of Gut Microbes and High-Fat Feeding on Host Circadian Clock Function and Metabolism

Effects of Diurnal Variation of Gut Microbes and High-Fat Feeding on Host Circadian Clock Function and Metabolism

A Gas Trapping Method for High-Throughput Metabolic Experiments

Hydrogen Sulfide Actions in the Vasculature

Contact Info

Product

Resources

About