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Single-nuclei RNA-Seq to characterize the immediate whole-animal postprandial transcriptional response following a high-fat meal in larval zebrafish

ABSTRACT: In this study, we employed a fluorescent BODIPY-fatty acid feeding assay and 10x Genomics single nuclei RNA-sequencing (snRNA-seq) to explore the simultaneous transcriptional responses following consumption of a high-fat meal in the context of a whole vertebrate organism (larval zebrafish). We used the whole-animal nuclear transcriptome sampling over short sampling periods (0 to 4 hours) to demonstrate how known metabolic pathways intertwine in new functional/spatial and immediate temporal contexts, and demonstrated the power of larval zebrafish snRNA-seq and fluorescent high-fat feeding protocols to describe the whole-animal response to consumption of a high-fat meal. To our knowledge, such an approach has never been accomplished before in a vertebrate model.

ORGANISM(S): Danio rerio

PROVIDER: GSE346015 | GEO | 2026/09/04

REPOSITORIES: GEO

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Project description:High-protein diets are known to reduce adiposity in the context of high carbohydrate and Western diets. However, few studies have investigated the specific high-protein effect on lipogenesis induced by a high-sucrose (HS) diet or fat deposition induced by high-fat feeding. We aimed to determine the effects of high protein intake on the development of fat deposition and partitioning in response to high-fat and/or HS feeding. A total of thirty adult male Wistar rats were assigned to one of the six dietary regimens with low and high protein, sucrose and fat contents for 5 weeks. Body weight (BW) and food intake were measured weekly. Oral glucose tolerance tests and meal tolerance tests were performed after 4th and 5th weeks of the regimen, respectively. At the end of the study, the rats were killed 2 h after ingestion of a calibrated meal. Blood, tissues and organs were collected for analysis of circulating metabolites and hormones, body composition and mRNA expression in the liver and adipose tissues. No changes were observed in cumulative energy intake and BW gain after 5 weeks of dietary treatment. However, high-protein diets reduced by 20 % the adiposity gain induced by HS and high-sucrose high-fat (HS-HF) diets. Gene expression and transcriptomic analysis suggested that high protein intake reduced liver capacity for lipogenesis by reducing mRNA expressions of fatty acid synthase (fasn), acetyl-CoA carboxylase a and b (Acaca and Acacb) and sterol regulatory element binding transcription factor 1c (Srebf-1c). Moreover, ketogenesis, as indicated by plasma β-hydroxybutyrate levels, was higher in HS-HF-fed mice that were also fed high protein levels. Taken together, these results suggest that high-protein diets may reduce adiposity by inhibiting lipogenesis and stimulating ketogenesis in the liver. Adult male Wistar rats were fed diets with varying amounts of protein, carbohydrates and fat for 5 weeks. At the end of the experiment, rats were killed and tanscriptome analysis was performed on pooled liver samples.

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Single-nuclei RNA-Seq to characterize the immediate whole-animal postprandial transcriptional response following a high-fat meal in larval zebrafish

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