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Updated: Jun 23, 2026
Laser Capture Microdissection of Mouse Embryonic Cartilage and Bone for Gene Expression Analysis
Published on: December 18, 2019
Localization of mRNAs and proteins in methyl methacrylate-embedded tissues
Jacob S Torgersen1, Harald Takle, Oivind Andersen
1NOFIMA Marin, PO Box 5010, N-1432 As, Norway. jacob.torgersen@nofima.no
Abstract:
Precise localization of proteins and mRNA in histological sections is necessary for evaluating spatial gene expression patterns. Here we report sensitive detection of the gene products in fish tissues by immunohistochemistry (IHC) and in situ hybridization (ISH) assays on sections of whole specimens and vertebra embedded in methyl methacrylate (MMA) resin. This plastic resin favors easy preparation of various specimen types and enables preparation of large sections with well-preserved cell morphology. IHC analysis of the muscle regulatory factor MyoD in transverse sections of juvenile cod revealed MyoD-positive cells in the dorsolateral parts of the adaxial muscle. ISH revealed less spatially restricted signals of the bone morphogenic protein bmp4 in muscle and brain. To assess the applicability of ISH on sections of bony tissue, col1a1 and col2a1 expression was investigated in non-decalcified vertebra sections of Atlantic salmon. The former was identified in both chondrocytes and osteoblasts, whereas the latter was mostly evident in chondrocytes. We conclude that MMA resin offers easy preparation of large and problematic tissues and the possibility of carrying out both IHC and ISH analyses using standard protocols.
Insights
Methyl methacrylate (MMA) resin enables sensitive detection of gene products in fish tissues using immunohistochemistry (IHC) and in situ hybridization (ISH). This method allows for precise spatial gene expression analysis in various tissue types, including bone.
Area of Science:
- Molecular Biology
- Histology
- Biotechnology
Background:
- Accurate spatial gene expression analysis requires precise protein and mRNA localization in histological sections.
- Standard histological techniques can be challenging for certain tissue types, including bony structures.
Purpose of the Study:
- To evaluate the utility of methyl methacrylate (MMA) resin for preparing fish tissues for molecular analyses.
- To demonstrate the effectiveness of immunohistochemistry (IHC) and in situ hybridization (ISH) on MMA-embedded sections.
Main Methods:
- Embedding whole fish specimens and vertebrae in methyl methacrylate (MMA) resin.
- Performing immunohistochemistry (IHC) for MyoD in juvenile cod muscle sections.
- Conducting in situ hybridization (ISH) for bmp4, col1a1, and col2a1 in fish tissue sections.
Main Results:
- MMA resin facilitated the preparation of large sections with well-preserved morphology.
- IHC successfully localized MyoD in specific muscle regions.
- ISH detected bmp4 in muscle and brain, and collagen genes (col1a1, col2a1) in salmon vertebrae, including chondrocytes and osteoblasts.
Conclusions:
- Methyl methacrylate (MMA) resin is a versatile embedding medium for preparing diverse and challenging fish tissues.
- MMA-embedded sections are suitable for sensitive IHC and ISH analyses, enabling robust spatial gene expression studies.
- This method supports the investigation of gene product localization in both soft and bony tissues using standard protocols.
