Identification of pro- and mature brain-derived neurotrophic factor in human saliva.
OBJECTIVE
Growth factors, including brain-derived neurotrophic factor (BDNF), are polypeptides that are involved in the maintenance, survival, and death of central and peripheral cells. Numerous growth factors have been identified in saliva and are thought to promote wound healing and maintenance of the oral epithelium. The aim of this study was to determine if BDNF is also found in human saliva.


METHODS
Whole, unstimulated saliva samples (n = 30) were analyzed by SDS-PAGE and Western blot using an anti-human BDNF antibody. Proteolytic cleavage products were similarly assessed following the incubation of pooled saliva with N-glycanase F and plasmin. Subjects were also genotyped for the BDNF Val66Met single nucleotide polymorphism (SNP).


RESULTS
These experiments revealed the presence of immunoreactive bands at 14, 32 and 34 kDa, corresponding to mature (mBDNF) and proBDNF, as well as a truncated pro-form at 24 kDa. Not every sample contained all forms of BDNF. Treatment with N-glycanase and plasmin reduced the size of the higher molecular weight bands, confirming the glycosylated pro-form of BDNF. mBDNF was detected significantly less often in subjects with the Val66Met SNP, compared to those without the polymorphism (chi(2) = 4.05; P < 0.05).


CONCLUSIONS
While the function of salivary BDNF still requires elucidation, these findings suggest that it may be possible to use saliva in lieu of blood in future studies of BDNF and the Val66Met polymorphism.
Mandel
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Monell Chemical Senses Center, Philadelphia, PA 19104, USA. amandel@monell.org
Ozdener
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Utermohlen
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D000328:Adult; D015153:Blotting, Western; D019208:Brain-Derived Neurotrophic Factor / Q000032:analysis* / Q000097:blood / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005341:Fibrinolysin; D005838:Genotype; D006026:Glycoside Hydrolases; D006801:Humans; D008297:Male; D008715:Methionine / Q000235:genetics; D008970:Molecular Weight; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D011498:Protein Precursors / Q000032:analysis* / Q000097:blood / Q000235:genetics; D012463:Saliva / Q000737:chemistry*; D014633:Valine / Q000235:genetics
D019208:Brain-Derived Neurotrophic Factor; D011498:Protein Precursors; C426674:brain-derived neurotrophic factor precursor; D008715:Methionine; D006026:Glycoside Hydrolases; C020668:glycanase; D005341:Fibrinolysin; D014633:Valine