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Animals : an open access journal from MDPI2025; 15(20); 2933; doi: 10.3390/ani15202933

Methodologies to Identify Metabolic Pathway Differences Between Emaciated and Moderately Conditioned Horses: A Review of Multiple Gene Expression Techniques.

Abstract: Starvation in horses presents critical welfare, economic, and management challenges with underlying molecular mechanisms of metabolic modification and recovery left poorly defined. Prolonged caloric deprivation induces significant systemic shifts in carbohydrate, protein, and lipid metabolism, reflected in coordinated changes in tissue-specific gene expression. This review synthesizes current knowledge on equine metabolic responses to starvation, emphasizing pathways found through RNA sequencing (RNA-seq) and real-time quantitative polymerase chain reaction (RT-qPCR) studies. Molecular investigations using RNA-seq and RT-qPCR have provided insight into transcriptional reprogramming during starvation and subsequent refeeding. Shifts in gene expression reflect the metabolic transition from carbohydrate dependence to lipid use, suppression of anabolic signaling, and activation of proteolytic pathways. However, interpretation of these data requires caution, as factors such as post-mortem interval, tissue handling, and euthanasia methods particularly the use of sodium barbiturates can influence transcript stability and abundance, potentially confounding results. The literature shows that starvation-induced molecular changes are not uniform across tissues, with skeletal muscle, liver, and adipose tissue showing distinct transcriptional signatures and variable recovery patterns during refeeding. Cross-species comparisons with hibernation, caloric restriction, and cachexia models provide context for understanding these changes, though equine-specific studies remain limited. Identified gaps include the scarcity of longitudinal data, inconsistent tissue sampling protocols, and lack of standardized reference genes for transcriptomic analyses in horses. Addressing these limitations will improve the accuracy of molecular evaluations and enhance our ability to predict recovery trajectories. A more comprehensive understanding of systemic and tissue-specific responses to starvation will inform evidence-based rehabilitation strategies, reduce the risk of refeeding syndrome, and improve survival and welfare outcomes for affected horses.
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Publication Date: 2025-10-10 PubMed ID: 41153862PubMed Central: PMC12560892DOI: 10.3390/ani15202933Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

Overview

  • This review article examines the molecular and genetic changes occurring in horses experiencing starvation compared to those in moderate condition.
  • It focuses on gene expression techniques, namely RNA sequencing (RNA-seq) and real-time quantitative polymerase chain reaction (RT-qPCR), to understand metabolic pathway shifts and recovery processes.

Introduction to the Research Context

  • Starvation in horses presents serious challenges relating to welfare, economics, and management.
  • The metabolic processes affected during starvation involve carbohydrate, protein, and lipid metabolism.
  • Changes during starvation are reflected in gene expression across different tissues, indicating systemic metabolic shifts.

Gene Expression Techniques Used

  • RNA sequencing (RNA-seq): A technique to quantify RNA levels across the entire transcriptome, providing a broad overview of gene expression changes.
  • Real-time quantitative polymerase chain reaction (RT-qPCR): A targeted approach to measure the relative expression levels of specific genes with high sensitivity.
  • Both methods have helped identify transcriptional reprogramming during starvation and refeeding phases.

Metabolic Pathway Findings

  • Starvation shifts metabolism:
    • From reliance on carbohydrates to increased lipid utilization for energy.
    • Anabolic pathways (responsible for growth and tissue building) are suppressed.
    • Proteolytic pathways (breaking down proteins for energy) are activated.
  • Gene expression patterns demonstrate these metabolic transitions across tissues.

Technical Considerations and Confounding Factors

  • Interpretations of gene expression data require caution due to several confounders:
    • Post-mortem interval – time between death and tissue sampling can alter transcript stability.
    • Tissue handling techniques – methods used in collecting and preserving tissues can affect RNA quality.
    • Euthanasia protocols – especially use of sodium barbiturates can influence transcript abundance and confound results.

Tissue-Specific Responses

  • Different tissues exhibit distinct gene expression responses to starvation:
    • Skeletal muscle: Shows changes related to increased protein breakdown and altered energy metabolism.
    • Liver: Exhibits transcriptional activity related to gluconeogenesis, lipid metabolism, and detoxification.
    • Adipose tissue: Reflects shifts in lipid mobilization and storage gene expression.
  • Recovery during refeeding varies by tissue, indicating different rates and mechanisms of metabolic normalization.

Comparative Context

  • Cross-species comparisons have been made with:
    • Hibernating animals, which undergo natural metabolic shifts.
    • Caloric restriction models.
    • Cachexia (wasting syndrome) in other species.
  • These comparisons help contextualize the equine-specific data but highlight the limited availability of focused equine studies.

Limitations and Gaps Identified

  • Scarcity of longitudinal studies tracking gene expression changes over time in the same animals.
  • Lack of standardized protocols for tissue sampling across different research groups.
  • Absence of consensus on reference genes for normalizing transcriptomic data specifically in horses.

Implications and Future Directions

  • Improving understanding of molecular responses to starvation will assist in designing better rehabilitation and management strategies.
  • Insights may reduce the risk of complications like refeeding syndrome by informing controlled nutritional recovery.
  • Future research needs include:
    • Developing standardized sampling and analysis protocols.
    • Conducting longitudinal and multi-tissue studies.
    • Establishing suitable reference genes for equine gene expression studies.
  • Ultimately, these advances will enhance welfare and survival outcomes for emaciated horses.

Cite This Article

APA
Austin MMP, Ivey JLZ, Shepherd EA, Myer PR. (2025). Methodologies to Identify Metabolic Pathway Differences Between Emaciated and Moderately Conditioned Horses: A Review of Multiple Gene Expression Techniques. Animals (Basel), 15(20), 2933. https://doi.org/10.3390/ani15202933

Publication

Animals : an open access journal from MDPI
ISSN: 2076-2615
NlmUniqueID: 101635614
Country: Switzerland
Language: English
Volume: 15
Issue: 20
PII: 2933

Researcher Affiliations

Austin, Madeline M P
  • Department of Animal Science, University of Tennessee, 2506 River Drive, Knoxville, TN 37996, USA.
Ivey, Jennie L Z
  • Department of Animal Science, University of Tennessee, 2506 River Drive, Knoxville, TN 37996, USA.
Shepherd, Elizabeth A
  • Department of Animal Science, University of Tennessee, 2506 River Drive, Knoxville, TN 37996, USA.
Myer, Phillip R
  • Department of Animal Science, University of Tennessee, 2506 River Drive, Knoxville, TN 37996, USA.

Conflict of Interest Statement

The authors declare no conflicts of interest.

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