Spontaneous oscillations in intracellular Ca(2+) concentration via purinergic receptors elicit transient cell swelling in rat parotid ducts.
Using multiphoton microscopy, we established that rat parotid ductal cells exhibit spontaneous oscillations in intracellular Ca(2+) concentration ([Ca(2+)](i)). These oscillatory Ca(2+) responses were observed during continuous perfusion with a physiological salt solution at 37 degrees C in the absence of calcium mobilizing agonist stimulation. The timing and patterns of these spontaneous Ca(2+) oscillations varied among individual ductal cells, and the average number of Ca(2+) responses in a single responding ductal cell was 2.1 in a 10-min recording period. High-speed scanning (0.6 s/image) revealed that most spontaneous elevations in [Ca(2+)](i) were initiated at the luminal side of ductal cells and spread toward the basal side within 2 s. Electron microscopic analysis after Ca(2+) imaging indicated that spontaneously oscillating ducts contained numerous granules at the luminal side, which is characteristic of granular ducts. These Ca(2+) oscillations were completely blocked by the purinergic receptor inhibitors 4-[[4-formyl-5-hydroxy-6-methyl-3-[(phosphonooxy)methyl]-2-pyridinyl]azo]-1,3-benzenedisulfonic acid (PPADS) and suramin but were not blocked by the muscarinic antagonist atropine or the alpha-adrenergic antagonist phentolamine. Simultaneous observation of fura-2 fluorescence and differential interference contrast (DIC) images showed that spontaneous elevations of [Ca(2+)](i) were well correlated with changes in shape of ductal cells. Using a plasma membrane fluorescence probe, SynaptoGreen C4, we found that the changes in DIC images reflected spontaneous cell swelling of ductal cells. Our findings present the possibility that purinergic receptors mediate spontaneous Ca(2+) oscillations in parotid ductal cells and regulate electrolyte reabsorption from the primary saliva in the resting state.
Shitara
Akiko
A
Dept. of Pharmacology, Health Sciences Univ. of Hokkaido, Japan.
Tanimura
Akihiko
A
Sato
Atsuko
A
Tojyo
Yosuke
Y
D000255:Adenosine Triphosphate / Q000378:metabolism; D000317:Adrenergic alpha-Antagonists / Q000494:pharmacology; D000818:Animals; D001285:Atropine / Q000494:pharmacology; D020013:Calcium Signaling / Q000187:drug effects; D048429:Cell Size / Q000187:drug effects; D005456:Fluorescent Dyes; D066298:In Vitro Techniques; D008297:Male; D046529:Microscopy, Electron, Transmission; D036641:Microscopy, Fluorescence, Multiphoton; D018727:Muscarinic Antagonists / Q000494:pharmacology; D010306:Parotid Gland / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D010646:Phentolamine / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D011983:Receptors, Purinergic / Q000187:drug effects / Q000378:metabolism*; D018987:Salivary Ducts / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D012815:Signal Processing, Computer-Assisted; D013498:Suramin / Q000494:pharmacology; D013997:Time Factors; D014544:Uridine Triphosphate / Q000378:metabolism
D000317:Adrenergic alpha-Antagonists; D005456:Fluorescent Dyes; D018727:Muscarinic Antagonists; D011983:Receptors, Purinergic; D013498:Suramin; D001285:Atropine; D000255:Adenosine Triphosphate; D014544:Uridine Triphosphate; D010646:Phentolamine