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Transcriptional and immunohistochemical signatures of transition dairy cows’ adipose tissue during subclinical ketosis

ABSTRACT: The objective of this experiment was to determine transcriptional differences in the visceral (VAT) and subcutaneous adipose tissue (SAT) of dairy cows with or without subclinical ketosis.

ORGANISM(S): Bos taurus

PROVIDER: GSE245350 | GEO | 2025/07/09

REPOSITORIES: GEO

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Project description:Adipose tissue plays a central role in regulating whole-body metabolic health, facilitated by the variety of cell types and their wide-ranging functions. In addition, depot-specific differences in adipose tissue have been shown to play important roles in different disease states including obesity, diabetes, and metabolic dysfunction in human and animal models. For early postpartum dairy cattle, metabolic dysfunction, triggered by a negative energy balance, is often manifested as subclinical ketosis (SCK). However, the role that subcutaneous (SAT) and visceral (VAT) adipose tissue depots, and their diverse cellular compositions, play in the response to subclinical ketosis conditions is unclear. Flank SAT and omental VAT were collected via laparotomy from five non-ketotic (NK; BHB ≤ 0.8 mmol/L) and five subclinical ketosis (SCK; 1.4 mmol/L < BHB ≤ 2.6 mmol/L) multiparous cows during early lactation. Following collection, nuclei were isolated from the tissue and subjected to single-nuclei RNA sequencing in order to investigate the transcriptional cellular heterogeneity. Distinct clusters of adipocytes (AD), adipose stem/progenitor cells (ASPC), immune cells (IMC), endothelial cells (EC), and pericyte/smooth muscle cells (PE/SMC) were identified in both adipose depots, with a greater abundance of ASPC in SAT compared to VAT. In addition, we identified a VAT-specific AD subtype characterized by higher expression of progenitor-like marker genes. While the abundance of none of the identified cell subtypes were different between SCK and NK, underlying transcriptional changes provided insight into potential effects of SCK. In general, SCK was associated with pro-lipogenic, anti-inflammatory, and pro-angiogenic transcriptional changes, possibly indicating a greater capacity for homeostatic responsiveness in SAT under conditions of enhanced negative energy balance. In contrast, SCK appeared to promote transcriptional changes indicative of impaired adipogenesis, impaired angiogenesis, and increased inflammation in VAT. Uniquely, our study presents novel insight into the cellular heterogeneity of adipose tissue in dairy cattle with subclinical ketosis. Furthering our understanding of the role of adipose tissue in response to this form of metabolic challenge has the potential to enhance efforts aimed at limiting the incidence and impact of subclinical ketosis and improving the health and productivity of dairy cattle.

Project description:Excessive body fat mobilization during the transition period overwhelms hepatic fatty acid (FA) oxidative capacity, leading to increased ketone body production and ketosis in dairy cows. Skeletal muscle (SM) has been shown to uptake and oxidize FA via mitochondrial β-oxidation and may play a crucial role in counteracting the hepatic FA overload. However, the role of SM in the pathogenesis of ketosis remains unclear. The objective of this study was to investigate transcriptomic alterations in the SM of postpartum dairy cows diagnosed with ketosis and identify genes and pathways potentially involved in the pathogenesis of the disease. Multiparous Holstein cows within 7 days in milk (DIM) were classified based on their blood β-hydroxybutyrate (BHB) concentrations as ketosis (BHB ≥ 1.3 mmol/L and reduced rumination; n = 15) and controls (BHB ≤ 1.1 mmol/L with no clinical signs; n = 15). On the day of enrollment, a biopsy from the external oblique muscle was collected and later processed for RNA sequencing. Differential expression analysis identified 3,543 differentially expressed genes (DEG), with 1,722 upregulated and 1,821 downregulated in the ketosis group relative to controls. Transcriptomic signatures of SM during ketosis revealed the upregulation of genes involved in protein synthesis, folding, and degradation, alongside downregulation of genes involved in mitochondrial energy production, extracellular matrix synthesis and remodeling, and muscle structure and contraction. In addition, our findings suggest impaired uptake and utilization of ketone bodies by SM in ketotic cows, as evidenced by the reduced expression of several genes involved in ketone catabolism. Although these findings suggest the involvement of SM in ketosis, further studies are warranted to determine whether these alterations contribute to the development and progression of ketosis or arise as a consequence of the disease. Keywords: ketosis, ketone bodies, RNA-Seq, skeletal muscle, muscle metabolism

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Transcriptional and immunohistochemical signatures of transition dairy cows’ adipose tissue during subclinical ketosis

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