In situ spatial transcriptomic analysis of human skeletal muscle using the Xenium platform.

Area of Science:

• Biotechnology
• Molecular Biology
• Muscle Physiology

Background:

• Traditional transcriptomics averages gene expression, masking skeletal muscle fibre type-specific differences.
• High-resolution spatial transcriptomics is needed to understand muscle heterogeneity.

Purpose of the Study:

• To evaluate the Xenium platform for spatial transcriptomic analysis of human skeletal muscle.
• To compare gene expression profiles between skeletal muscle fibre types (Type 1 and Type 2).

Main Methods:

• Human vastus lateralis muscle sections were analyzed using the Xenium platform.
• Myosin Heavy Chain isoform staining differentiated muscle fibre types.
• Manual segmentation enabled accurate fibre type-specific transcript density comparisons.

Main Results:

• Xenium platform successfully performed high-resolution spatial transcriptomics on muscle sections.
• Type 1 fibres showed higher overall transcript density, especially in nuclear regions.
• 191 genes were differentially expressed between muscle fibres and perimysium, with specific genes identified for Type 1 and Type 2 fibres.

Conclusions:

• The Xenium platform, with manual segmentation, enables effective fibre type-specific transcriptomic analysis in skeletal muscle.
• This approach provides novel insights into the molecular distinctions between human skeletal muscle fibre types.