Abstract
Modern approaches to discovering molecular mechanisms and validating treatments for age-related neuromusculoskeletal dysfunction typically rely on high-throughput transcriptome analysis. Previously harvested and fixed tissues offer an incredible reservoir of untapped molecular information. However, obtaining RNA from such formaldehyde-fixed neuromusculoskeletal tissues, especially fibrotic aged tissues, is technically challenging and often results in RNA degradation, chemical modification and yield reduction, prohibiting further analysis. Therefore, we developed a protocol to extract high-quality RNA from formaldehyde-fixed brain, cartilage, muscle and peripheral nerve isolated from naturally aged mice. Isolated RNA produced reliable gene expression data comparable to fresh and flash-frozen tissues and was sensitive enough to detect age-related changes, making our protocol valuable to researchers in the field of aging.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 153-160 |
| Number of pages | 8 |
| Journal | BioTechniques |
| Volume | 76 |
| Issue number | 4 |
| DOIs | |
| State | Published - 2024 |
Funding
This work was supported by the Lisa Dean Moseley Foundation, the Frankel Family Foundation, a Shirley Ryan AbilityLab Innovative Catalyst Grant and the NIH National Institute on Aging (R01 AG073223-01A1) (M Lavasani) as well as the US Department of Veterans Affairs Research and Development Service (IK6 RX003351) (RL Lieber). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed. Competing interests disclosure
Keywords
- RNA isolation from fixed tissue
- aging
- formaldehyde fixation
- gene expression
- neuromusculoskeletal tissues
- tissue bank
ASJC Scopus subject areas
- Biotechnology
- General Biochemistry, Genetics and Molecular Biology