A Novel System of Human Submandibular/Sublingual Saliva Collection

Wolff, Ändy; Begleiter, Asher; Moskona, D

doi:10.1177/00220345970760111001

Cited by 54 publications

(30 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Saliva secretion was induced in anesthetized mice by pilocarpine and collected using a minisuction apparatus based on a recent design (23). In wild-type mice, marked salivation was observed within 20 s after pilocarpine injection, and substantial amounts of clear, nonviscous fluid were collected.…”

Section: Resultsmentioning

confidence: 99%

Defective Secretion of Saliva in Transgenic Mice Lacking Aquaporin-5 Water Channels

Song

Gillespie

et al. 1999

Journal of Biological Chemistry

506

440

View full text Add to dashboard Cite

Aquaporin-5 (AQP5) is a water-selective transporting protein expressed in epithelial cells of serous acini in salivary gland. We generated AQP5 null mice by targeted gene disruption. The genotype distribution from intercross of founder AQP5 heterozygous mice was 70: 69:29 wild-type:heterozygote:knockout, indicating impaired prenatal survival of the null mice. The knockout mice had grossly normal appearance, but grew ϳ20% slower than litter-matched wild-type mice when placed on solid food after weaning. Pilocarpine-stimulated saliva production was reduced by more than 60% in AQP5 knockout mice. Compared with the saliva from wildtype mice, the saliva from knockout mice was hypertonic (420 mosM) and dramatically more viscous. Amylase and protein secretion, functions of salivary mucous cells, were not affected by AQP5 deletion. Water channels AQP1 and AQP4 have also been localized to salivary gland; however, pilocarpine stimulation studies showed no defect in the volume or composition of saliva in AQP1 and AQP4 knockout mice. These results implicate a key role for AQP5 in saliva fluid secretion and provide direct evidence that high epithelial cell membrane water permeability is required for active, near-isosmolar fluid transport.The family of molecular water channels (aquaporins) numbers 10 in mammals and many more in plants and lower organisms. There has been considerable recent interest in the role of aquaporins in mammalian physiology and disease mechanisms. In humans, mutation of the vasopressin-regulated water channel of kidney collecting, AQP2, 1 causes hereditary nephrogenic diabetes insipidus in which patients are unable to concentrate their urine (1). Recent phenotype characterization of transgenic knockout mice lacking AQP1 and AQP4 has been very informative in defining the roles of these water channels in the urinary concentrating mechanism, lung fluid transport, and gastrointestinal physiology (2-6). However the phenotype studies indicated that the tissue expression of an aquaporin does not ensure its functional significance.AQP5 is a water channel with a unique tissue expression pattern (7). Immunocytochemical studies from several laboratories showed AQP5 expression in the apical membranes of serous acinar cells in salivary and lacrimal glands, type I alveolar epithelial cells, and surface corneal epithelial cells (8 -11). AQP5 appears to function as an unregulated waterselective channel with comparable intrinsic water permeability to AQP1 (12). The human AQP5 gene contains 4 exons with exon-intron boundaries at identical locations to those several other aquaporins (13); the genes for AQP5, AQP2, and AQP6 are clustered in a small 27-kb region at chromosome locus 12q13 (14). It was proposed that AQP5 plays an important role in glandular secretions of saliva and tears and that abnormalities in AQP5 might occur in some forms of Sjogren's syndrome (15,16). Aquaporin gene delivery to salivary gland has been proposed to increase fluid secretion (15). However, these possibilities are based on the unproven assu...

show abstract

Section: Resultsmentioning

confidence: 99%

Defective Secretion of Saliva in Transgenic Mice Lacking Aquaporin-5 Water Channels

Song

Gillespie

et al. 1999

Journal of Biological Chemistry

506

440

View full text Add to dashboard Cite

show abstract

“…Donors were in good health and exhibited normal salivary function. Parotid saliva secretions were harvested using a saliva collector [17] fitted with a sterile 100 L pipette tip. Stimulation of the salivary glands was provided by repeated topical application of a mild solution of citric acid (2%) to the dorsal surface of the tongue.…”

Section: Sample Collectionmentioning

confidence: 99%

Micro-heterogeneity of human saliva peptide P-C characterized by high-resolution top-down fourier-transform mass spectrometry

Halgand

Zabrouskov

Bassilian

et al. 2010

J. Am. Soc. Mass Spectrom.

View full text Add to dashboard Cite

Top-down proteomics characterizes protein primary structures with unprejudiced descriptions of expressed and processed gene products. Gene sequence polymorphisms, protein post-translational modifications, and gene sequence errors can all be identified using topdown proteomics. Saliva offers advantages for proteomic research because of availability and the noninvasiveness of collection and, for these reasons, is being used to search for disease biomarkers. The description of natural protein variants, and intra-and inter-individual polymorphisms, is necessary for a complete description of any proteome, and essential for the discovery of disease biomarkers. Here, we report a striking example of natural protein variants with the discovery by top-down proteomics of two new variants of Peptide P-C. Intact mass measurements, and collisionally activated-, infrared multiphoton-, and electron capturedissociation, were used for characterization of the form predicted from the gene sequence with an average mass 4371 Da, a form postulated to result from a single nucleotide polymorphism of mass 4372 Da, and another form of mass 4370 Da postulated to arise from a novel protein sequence polymorphism. While the biological significance of such subtle variations in protein structure remains unclear, their importance cannot be assigned without their characterization, as is reported here for one of the major salivary proteins. (J Am Soc Mass Spectrom 2010, 21, 868 -877)

show abstract

“…Parotid saliva was collected as described by Lashley (20 ), using a Lashleylike cup that was fabricated by the UCLA Life Science Machine Shop. Submandibular and sublingual saliva collections were obtained by use of a Wolff collector according to the published protocol (21,22 ). GCF was collected with the paper strip sampling technique with minor modifications (23 ).…”

Section: Saliva and Blood Collection And Rna Isolationmentioning

confidence: 99%

Characterization of RNA in Saliva

et al. 2006

View full text Add to dashboard Cite

Background:We have previously shown that human mRNAs are present in saliva and can be used as biomarkers of oral cancer. In this study, we analyzed the integrity, sources, and stability of salivary RNA. Methods: We measured the integrity of salivary RNA with reverse transcription followed by PCR (RT-PCR) or RT-quantitative PCR (RT-qPCR). To study RNA entry sites into the oral cavity, we used RT-PCR analysis of salivary RNA from the 3 major salivary glands, gingival crevice fluid, and desquamated oral epithelial cells. We measured stability of the salivary ␤-actin mRNA by RT-qPCR of salivary RNA incubated at room temperature for different periods of time. We measured RNA association with other macromolecules by filtering saliva through pores of different sizes before performing RT-qPCR. To assess RNA-macromolecule interaction, we incubated saliva with Triton X-100 for different periods of time before performing RT-qPCR. Results: In most cases, we detected partial-to fulllength salivary mRNAs and smaller amounts of middle and 3 gene amplicons compared with the 5. RNA was present in all oral fluids examined. Endogenous salivary ␤-actin mRNA degraded more slowly than exogenous ␤-actin mRNA, with half-lives of 12.2 and 0.4 min, respectively (P <0.001). Salivary RNA could not pass through 0.22 or 0.45 m pores. Incubation of saliva with Triton X-100 accelerated degradation of salivary RNA. Conclusions: Saliva harbors both full-length and partially degraded forms of mRNA. RNA enters the oral cavity from different sources, and association with macromolecules may protect salivary RNA from degradation.

show abstract

A Novel System of Human Submandibular/Sublingual Saliva Collection

Cited by 54 publications

References 21 publications

Defective Secretion of Saliva in Transgenic Mice Lacking Aquaporin-5 Water Channels

Defective Secretion of Saliva in Transgenic Mice Lacking Aquaporin-5 Water Channels

Micro-heterogeneity of human saliva peptide P-C characterized by high-resolution top-down fourier-transform mass spectrometry

Characterization of RNA in Saliva

Contact Info

Product

Resources

About