A rat parotid gland cell line, Par-C10, exhibits neurotransmitter-regulated transepithelial anion secretion

Abstract

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 ( I sc) in response to relevant receptor agonists. Par-C10 cultures reached confluence in 3–4 days and developed transcellular resistance values of ≥2,000 Ω ⋅ 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 I sc was increased in response to muscarinic cholinergic and α- and β-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 I sc. 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.

P2Y2 Nucleotide Receptors Enhance α-Secretase-dependent Amyloid Precursor Protein Processing

“…UTP also caused a dose-dependent increase in sAPP␣ from 1321N1-P2Y 2 cells with an EC 50 value of 2.5 M (Fig. 2), which is characteristic of P2Y 2 R activation (5,6,53,54). In contrast, UTP-induced sAPP␣ release was undetectable in untransfected cells or in cells transfected with the expression vector only (data not shown).…”

“…Several physiological responses resulting from P2Y 2 R activation are dependent on the rise of [Ca 2ϩ ] i (e.g. ion secretion, fluid secretion, and vascular cell adhesion molecule-1 expression) (21,54,68); however, our results indicate that P2Y 2 R-mediated increases in [Ca 2ϩ ] i by either calcium released from intracellular stores or calcium influx were not required for sAPP␣ release, because chelation of intracellular calcium with BAPTA had no effect on UTP-induced sAPP␣ release (Fig. 4A).…”

“…In addition, P2Y 2 Rs can promote calcium entry through the stimulation of plasma membrane calcium channels (5,13,54,55). Therefore, we examined the role of calcium signaling pathways in UTP-induced sAPP␣ release.…”

P2Y2 nucleotide receptor activation enhances the aggregation and self-organization of dispersed salivary epithelial cells

“…In the following studies, we used the rat parotid acinar (Par-C10) cell line, an established in vitro model of salivary gland differentiation and function (7,112). Par-C10 cells express endogenous P2Y 2 Rs, but not P2Y 4 R (unpublished observations) or P2Y 6 R (112).…”

A systems biology approach identifies a gene regulatory network in parotid acinar cell differentiation.

“…These cells also express AQP5, they contain electron dense granules, and increased expression of amylase [62]. Similarly, ParC10 cells (a cell line derived from primary rat parotid acinar cells, and immortalized with simian virus 40 (SV40) [63,64] also organize into lumenized spheres on matrigel, express apically localized TJ proteins, basally localized muscarinic receptor, and increased expression of AQP5 [65].…”