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Laser Capture Microdissection of Mouse Embryonic Cartilage and Bone for Gene Expression Analysis

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Published on: December 18, 2019

Quantitative analysis of mRNA in human temporal bones.

Yurika Kimura¹, Sachiho Kubo, Hiroko Koda

¹Department of Otolaryngology, Tokyo Metropolitan Geriatric Hospital, Japan.

Acta Oto-Laryngologica

|September 14, 2007

Summary

Frozen human inner ear tissue yields well-preserved messenger RNA (mRNA), enabling molecular studies of inner ear disorders. This method is crucial for understanding disease mechanisms in human specimens.

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Area of Science:

Otolaryngology
Molecular Biology
Human Genetics

Background:

Studying the molecular mechanisms of inner ear disorders requires RNA and DNA analysis.
Established methods for RNA isolation exist for animal models, but human inner ear RNA isolation is challenging.

Purpose of the Study:

To demonstrate a method for extracting and quantitatively analyzing messenger RNA (mRNA) from human inner ears.
To establish a protocol for molecular analysis of human inner ear tissues.

Main Methods:

Messenger RNA (mRNA) was extracted from human inner ear tissues (membranous labyrinths) obtained from both frozen and formalin-fixed temporal bones.
Quantitative RT-PCR was used to compare the length and quantity of COCH mRNA and GAPDH mRNA between frozen and formalin-fixed specimens.

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Main Results:

Well-preserved COCH mRNA (up to 976 bp) was successfully amplified from all three frozen specimens.
Formalin-fixed specimens yielded significantly less amplifiable COCH mRNA (249 bp) and only 1% of the GAPDH mRNA quantity compared to frozen specimens.

Conclusions:

Frozen human inner ear tissue is suitable for extracting well-preserved mRNA.
This method allows for the molecular study of inner ear disorders using human specimens.