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1 Department of Pharmacology, School of Medicine, University of Missouri, Columbia, Missouri 65212; 2 Oral Pathology Research Laboratory, Department of Veterans Affairs Medical Center, Washington, District of Columbia 20422; and 3 Department of Basic Sciences and Oral Research, School of Dentistry, University of Colorado Health Sciences Center, Denver, Colorado 80262
Because of the
lack of salivary gland cell lines suitable for Ussing chamber studies,
a recently established rat parotid acinar cell line, Par-C10, was grown
on permeable supports and evaluated for development of transcellular
resistance, polarization, and changes in short-circuit current
(Isc) in
response to relevant receptor agonists. Par-C10 cultures reached
confluence in 3-4 days and developed transcellular resistance
values of
2,000
· cm2.
Morphological examination revealed that Par-C10 cells grew as polarized
monolayers exhibiting tripartite junctional complexes and the acinar
cell-specific characteristic of secretory canaliculi. Par-C10
Isc was increased
in response to muscarinic cholinergic and
- and
-adrenergic
agonists on the basolateral aspect of the cultures and to ATP and UTP
(through P2Y2 nucleotide
receptors) applied apically. Ion replacement and inhibitor studies
indicated that anion secretion was the primary factor in
agonist-stimulated Isc. RT-PCR,
which confirmed the presence of
P2Y2 nucleotide receptor mRNA in
Par-C10 cells, also revealed the presence of mRNA for the cystic
fibrosis transmembrane conductance regulator and ClC-2 Cl
channel proteins. These findings
establish Par-C10 cells as the first cell line of salivary gland origin
useful in transcellular ion secretion studies in Ussing chambers.
parotid salivary gland; cell culture; Ussing chamber; ion secretion; P2Y2 receptors; polarized epithelia
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