Convenient and reproducible in vivo gene transfer to mouse parotid glands.
OBJECTIVE
Published studies of gene transfer to mouse salivary glands have not employed the parotid glands. Parotid glands are the likely target tissue for most clinical applications of salivary gene transfer. The purpose of the present study was to develop a convenient and reproducible method of retroductal gene transfer to mouse parotid glands.


METHODS
The volume for vector delivery was assessed by infusion of Toluidine Blue into Stensen's ducts of Balb/c mice after direct intraoral cannulation. Recombinant, serotype 5 adenoviral vectors, encoding either firefly luciferase or human erythropoietin (hEpo), were constructed and then administered to parotid glands (10(7) vector particles/gland). Transgene expression in vivo was measured by enzyme activity (luciferase) or an enzyme-linked immunosorbent assay (hEpo). Vector biodistribution was measured by real-time quantitative (Q) PCR.


RESULTS
The chosen volume for mouse parotid vector delivery was 20μL. Little vector was detected outside of the targeted glands, with both QPCR and luciferase assays. Transgene expression was readily detected in glands (luciferase, hEpo), and serum and saliva (hEpo). Most secreted hEpo was detected in saliva.


CONCLUSIONS
These studies show that mouse parotid glands can be conveniently and reproducibly targeted for gene transfer, and should be useful for pre-clinical studies with many murine disease models.
Zheng
C
C
Molecular Physiology and Therapeutics Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD 20892-1190, USA.
Shinomiya
T
T
Goldsmith
C M
CM
Di Pasquale
G
G
Baum
B J
BJ
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D004921:Erythropoietin / Q000008:administration & dosage / Q000235:genetics / Q000378:metabolism*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D008156:Luciferases / Q000008:administration & dosage / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D023421:Models, Animal; D009928:Organ Specificity; D010306:Parotid Gland / Q000378:metabolism*; D011994:Recombinant Proteins / Q000008:administration & dosage; D012463:Saliva / Q000378:metabolism; D012471:Salivary Proteins and Peptides / Q000032:analysis / Q000378:metabolism
D011994:Recombinant Proteins; D012471:Salivary Proteins and Peptides; D004921:Erythropoietin; D008156:Luciferases