Which housekeeping gene is the best choice for RT-qPCR analysis in mice fed with a high-fat diet? Studies in the liver, kidney, pancreas, and intestines

Secio-Silva, Ayla; Emrich, Felipe; Evangelista-Silva, Paulo H.; Prates, Rodrigo Pereira; Hijo, Andressa Harumi Torelli; Figueira-Costa, Tatienne Neder; Schaeffer, Marie; Goulart-Silva, Francemilson; Peliciari-Garcia, Rodrigo A.; Bargi‐Souza, Paula

doi:10.1016/j.genrep.2023.101756

Cited by 6 publications

(1 citation statement)

References 33 publications

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“…The primers used are listed in Table . GAPDH was the reference gene used in our study. − Samples with RNAs that were not reverse transcribed and samples without RNA and cDNA served as negative controls. The relative abundance of each gene was calculated using the 2 –ΔΔ Ct method …”

Section: Methodsmentioning

confidence: 99%

Natural Sweetener Glycyrrhetinic Acid Monoglucuronide Improves Glucose Homeostasis in Healthy Mice

Wang,

Guo,

Zheng

et al. 2024

J. Agric. Food Chem.

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Noncaloric or low-caloric sweeteners have become popular worldwide, although debates persist regarding their impact on health. To investigate whether the sweeteners are favorable for glucose homeostasis, our study assessed the effects of glycyrrhetinic acid monoglucuronide (GAMG) and several commonly used sweeteners [glycyrrhetinic acid (GA), stevioside, erythritol, sucralose, and aspartame] on glycometabolism and elucidated the underlying mechanisms. The C57BL/6J male mice were exposed to different sweeteners for 10 weeks, and our results showed that GAMG significantly reduced fasting blood glucose (FBG) levels (FBG-control: 3.81 ± 0.42 mmol/L; FBG-GAMG: 3.37 ± 0.38 mmol/L; p < 0.05) and the blood glucose levels 15 and 30 min after sucrose or maltose loading (p < 0.05). Furthermore, it improved glucose tolerance (p = 0.028) and enhanced insulin sensitivity (p = 0.044), while the other sweeteners had negligible or adverse effects on glucose homeostasis. Subsequent experiments showed that GAMG inhibited α-glucosidases potently (IC 50 = 0.879 mg•mL −1 ), increased three SCFA-producing bacteria and SCFAs levels (p < 0.05), and promoted the gene expression of SCFA receptor GPR43 (p = 0.018). These results suggest that GAMG may regulate blood glucose by inhibiting α-glucosidases and modulating gut microbial SCFAs. Our findings prove that GAMG, beneficial to blood glucose regulation, is a promising natural sweetener for future utilization.

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Section: Methodsmentioning

confidence: 99%

Natural Sweetener Glycyrrhetinic Acid Monoglucuronide Improves Glucose Homeostasis in Healthy Mice

Wang,

Guo,

Zheng

et al. 2024

J. Agric. Food Chem.

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Biotic and abiotic stress affects the stability of housekeeping genes in Catla Labeo catla

Acharya,

Tripathi,

Rasal

et al. 2024

J Aqua Anim Hlth

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ObjectiveThe choice of a stable reference gene plays a crucial role in gene expression analysis. In most cases, the choice of housekeeping gene (HKG) is quite random. However, the expression of HKGs varies under different experimental conditions and in different tissues.MethodsThe study focused on three reference genes in Catla Labeo catla—beta actin (β‐actin), glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH), and elongation factor‐1 alpha (EF‐1α)—under four experimental conditions: bacterial challenge, hypoxia, a combination of bacterial challenge and hypoxia, and control. The fish were subjected to the above‐mentioned experimental circumstances, and five tissues (blood, gill, liver, kidney, and spleen) were collected at regular intervals.ResultThe investigation of gene expression in the five tissues was performed using real‐time (RT) quantitative polymerase chain reaction (qPCR), and the resultant cycle threshold values were subjected to several statistical techniques via the RefFinder tool to determine the most stable HKG. The results were ordered in accordance with the stability of the reference genes. In this study, GAPDH was found to be the least stable gene, while β‐actin and EF‐1α remained stable in the kidney and liver under various circumstances. Other tissues demonstrated tissue‐ and treatment‐specific stability for the various HKGs.ConclusionThe findings indicate that GAPDH should be avoided in Catla for gene expression studies and that different algorithms should be employed to validate the stability of HKGs in RT‐qPCR analysis.

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A systematic review on the selection of reference genes for gene expression studies in rodents: are the classics the best choice?

Bunde,

Pedra,

de Oliveira

et al. 2024

Mol Biol Rep

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Which housekeeping gene is the best choice for RT-qPCR analysis in mice fed with a high-fat diet? Studies in the liver, kidney, pancreas, and intestines

Cited by 6 publications

References 33 publications

Natural Sweetener Glycyrrhetinic Acid Monoglucuronide Improves Glucose Homeostasis in Healthy Mice

Natural Sweetener Glycyrrhetinic Acid Monoglucuronide Improves Glucose Homeostasis in Healthy Mice

Biotic and abiotic stress affects the stability of housekeeping genes in Catla Labeo catla

A systematic review on the selection of reference genes for gene expression studies in rodents: are the classics the best choice?

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