Validating drg 945
The results were expressed as number of cells per section.To avoid double counting, we have chosen every third DRG section for BDNF immunostaining.For analysis of the TRPV1 immunoreactivity in the distal colon, three random microscopy fields were chosen from each section with caution to avoid field overlap.Each field was positioned to view a similar morphological area covering the entire width of the colon that showed positive TRPV1 stain.The histology of the colon and the urinary bladder was examined by hematoxylin and eosin (H&E) stain.The protein expression of transient receptor potential (TRP) ion channel of the vanilloid type 1 (TRPV1) and brain-derived neurotrophic factor (BDNF) were examined by immunohistochemistry and/or western blot.The specificity of the antibodies was also evaluated with western blot or pre-absorption assay.Tissue sections were viewed and analyzed under a Zeiss fluorescent photomicroscope.
These observations indicate a broad phenomenon between organ to organ sensory cross-interaction.
DRG neurons expressing BDNF were counted in 4 to 6 sections of each DRG, and expressed as mean ± SE for n animals.
Similar sized sections were chosen using the microscope measurement program and all the positive cells were counted in the sections.
Studies with animal models have demonstrated that activation of primary afferent pathways may have a role in mediating viscero-visceral cross-organ sensitization.
Colonic inflammation was induced by a single dose of tri-nitrobenzene sulfonic acid (TNBS) instilled intracolonically.