Neuropeptide immunofluorescence in human nasal mucosa: assessment of the technique for vasoactive intestinal peptide (VIP).
Neuropeptides are important neurotransmitters in nasal physiology and the increasing knowledge of their role in nasal diseases brings new therapeutic perspectives. The investigation of human nasal mucosa neuropeptides is based mostly on immunocytochemistry, a complex approach whose resulting factors may be variable. Aiming to make this kind of research available, an immunofluorescence approach for vasoactive intestinal peptide (VIP) in human nasal mucosa is proposed and evaluated.


METHODS
Transversal cohort.


METHODS
Human inferior turbinate samples were obtained at time of nasal surgery from eight patients. The samples were fixed in Zamboni solution (4% phosphate-buffered paraformaldehyde and 0.4% picric acid), snap-frozen and stored at -70 degrees C. 14 microm sections were then obtained. Immunofluorescence staining for VIP (Peninsula Laboratories) was performed and its images documented by conventional photography. The method's specificity, sensitivity and reproducibility of execution were evaluated. Additionally, the reproducibility of interpretation of results was evaluated through the comparison of staining scores (0 to 4) attributed to the images by six observers.


RESULTS
The results showed the approach to be very specific and sensible, besides being reproducible in its execution. The interpretation of results may depend on the observer's accuracy in judging immunofluorescence images, but it showed uniformity.


CONCLUSIONS
The proposed method was highly useful for research purposes in neuropeptides in human nasal mucosa.
Mendonça
Jeferson Cedaro de
JC
Department of Otorhinolaryngology, Santa Casa de Sao Paulo, Faculdade de Ciências Médicas.
Dolci
José Eduardo Lutaif
JE
D003430:Cross-Sectional Studies; D005455:Fluorescent Antibody Technique / Q000379:methods*; D006801:Humans; D009297:Nasal Mucosa / Q000737:chemistry*; D009479:Neuropeptides / Q000032:analysis / Q000276:immunology; D012680:Sensitivity and Specificity; D014420:Turbinates / Q000737:chemistry; D014660:Vasoactive Intestinal Peptide / Q000032:analysis*
D009479:Neuropeptides; D014660:Vasoactive Intestinal Peptide