Functional and molecular characterization of the fluid secretion mechanism in human parotid acinar cells.
The strategies available for treating salivary gland hypofunction are limited because relatively little is known about the secretion process in humans. An initial microarray screen detected ion transport proteins generally accepted to be critically involved in salivation. We tested for the activity of some of these proteins, as well as for specific cell properties required to support fluid secretion. The resting membrane potential of human acinar cells was near -51 mV, while the intracellular [Cl-] was approximately 62 mM, about fourfold higher than expected if Cl ions were passively distributed. Active Cl- uptake mechanisms included a bumetanide-sensitive Na+ -K+ -2Cl- cotransporter and paired DIDS-sensitive Cl-/HCO3- and EIPA-sensitive Na+/H+ exchangers that correlated with expression of NKCC1, AE2, and NHE1 transcripts, respectively. Intracellular Ca2+ stimulated a niflumic acid-sensitive Cl- current with properties similar to the Ca2+ -gated Cl channel BEST2. In addition, intracellular Ca2+ stimulated a paxilline-sensitive and voltage-dependent, large-conductance K channel and a clotrimazole-sensitive, intermediate-conductance K channel, consistent with the detection of transcripts for KCNMA1 and KCNN4, respectively. Our results demonstrate that the ion transport mechanisms in human parotid glands are equivalent to those in the mouse, confirming that animal models provide valuable systems for testing therapies to prevent salivary gland dysfunction.
Nakamoto
Tetsuji
T
The Center for Oral Biology in the Aab Institute of Biomedical Sciences, University of Rochester School of Medicine and Dentistry, Rochester, New York 14642, USA.
Srivastava
Alaka
A
Romanenko
Victor G
VG
Ovitt
Catherine E
CE
Perez-Cornejo
Patricia
P
Arreola
Jorge
J
Begenisich
Ted
T
Melvin
James E
JE
D000328:Adult; D000368:Aged; D000818:Animals; D001826:Body Fluids / Q000378:metabolism*; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D007473:Ion Channels / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D010306:Parotid Gland / Q000378:metabolism*; D014882:Water-Electrolyte Balance / Q000502:physiology*
D007473:Ion Channels