Protocol for the isolation of silk glands from silkworms for snRNA-seq and spatial transcriptomics

Area of Science:

• Molecular Biology
• Genomics
• Entomology

Background:

• Silk glands (SGs) in silkworms are crucial for silk production, directly impacting silk yield and quality.
• Understanding the cellular and molecular mechanisms within SGs is key to improving silk properties.

Purpose of the Study:

• To develop and present a detailed protocol for isolating silk gland nuclei from silkworms.
• To enable high-quality tissue sectioning for spatial transcriptomics analysis of silk glands.

Main Methods:

• The protocol covers silkworm rearing, dissection, and nucleus isolation.
• It includes procedures for tissue embedding, cryosectioning, and RNA capture for sequencing.

Main Results:

• The described protocol yields high-quality tissue slices suitable for spatial transcriptomics.
• This approach facilitates the investigation of silk gland cell distribution at the single-cell level.

Conclusions:

• The presented protocol is effective for spatial transcriptomics of silkworm silk glands.
• It offers a valuable tool for exploring cellular heterogeneity and gene expression patterns within SGs.