A rat parotid gland cell line, Par-C10, exhibits neurotransmitter-regulated transepithelial anion secretion
A rat parotid gland cell line, Par-C10, exhibits neurotransmitter-regulated transepithelial anion secretion
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DOI：
10.1152/ajpcell.1998.275.2.c367
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发表时间：
1998-08-01
影响因子：
5.5
通讯作者：
Quissell, DO
中科院分区：
文献类型：
--
作者：
Turner, JT;Redman, RS;Quissell, DO
由于缺乏适合于Ussing小室研究的唾液腺细胞系，最近建立的大鼠腮腺腺泡细胞系PAR-C10被培养在可渗透的载体上，并对其跨细胞阻力、极化和短路电流(I-sc)对相关受体激动剂的反应进行了评估。PAR-C10培养在3-4天内达到融合，并产生大于或等于2,000欧米卡·厘米(2)的跨细胞阻力值。形态观察显示，PAR-C10细胞呈极化单层生长，呈现三部分连接复合体和腺泡细胞特有的分泌小管特征。PAR-C10I-sc在培养的基底侧受到M胆碱能、α和β肾上腺素能激动剂以及顶端施加的ATP和UTP(通过P2Y(2)核苷酸受体)的影响而增加。离子替代和抑制剂研究表明，阴离子分泌是激动剂刺激的i-sc的主要因素。RT-PCR证实Par-C10细胞中存在P2Y(2)核苷酸受体mRNA，同时也发现囊性纤维化跨膜电导调节蛋白和ClC-2Cl-通道蛋白的mRNA存在。这些发现确立了PAR-C10细胞是第一个唾液腺来源的细胞系，可用于Ussing小室的跨细胞离子分泌研究。
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 greater than or equal to 2,000 Omega.cm(2). 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 alpha- and beta-adrenergic agonists on the basolateral aspect of the cultures and to ATP and UTP (through P2Y(2) 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 P2Y(2) 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.