Genotyping and Characterisation of Transgenic Mice
In a Rheumatoid Arthritis Model

Genotyping and Characterisation of Transgenic Mice
In a Rheumatoid Arthritis Model

Rheumatoid arthritis and the receptors, molecules and immune cells involved in its pathophysiology.

Ekom Akanimo

2025

Rheumatoid arthritis (RA) is a chronic autoimmune disease characterised by persistent synovial inflammation, joint destruction, and pain. While immune mechanisms underlying RA are well established, emerging evidence highlights an important role for neuro-immune crosstalk in disease progression. In particular, mast cells and transient receptor potential (TRP) ion channels may contribute to inflammatory signalling within the synovium. This study aimed to genotype and characterise transgenic mouse models of RA to investigate the roles of mast cells and TRP channels, with a focus on TRPM2 and TRPM8. Wild-type, heterozygous, conditional knockout, and global knockout mice were analysed. Genotyping was performed using PCR and agarose gel electrophoresis to confirm genotype. Histological analysis of mouse knee joints was conducted using haematoxylin and eosin staining to assess tissue morphology, toluidine blue staining to identify mast cells, and immunostaining to detect TRP ion channel expression. Protein analysis was performed using ELISA to quantify inflammatory mediators, including IL-6, PGE₂, and Substance P, alongside Bradford assays to determine total protein content. PCR successfully confirmed the generation of transgenic mouse lines. Histological analysis revealed synovial inflammation and mast cell infiltration characteristic of RA, while immunostaining identified TRPM2 and TRPM8 expression within inflamed joints. TRPM2 knockout mice exhibited reduced inflammatory features compared to wild-type controls. ELISA confirmed the presence of key inflammatory mediators, supporting a mechanistic link between neuronal signalling and mast cell-driven inflammation. Collectively, these findings support a contributory role for mast cells and TRP ion channels in RA pathology and highlight their potential as therapeutic targets.

Biology, Neuroscience, Science, Biomedicine