Amylase secretion by cultured porcine parotid cells.
The similarity of porcine and human physiology and the availability of slaughterhouse tissues suggests the use of porcine parotid cells as a model for amylase secretion. A procedure is described for the isolation of porcine parotid cells by collagenase-P/dispase digestion of the tissue. The preparation consisted of individual cells and small aggregates that were maintained in primary culture, during which the cells formed aggregates that firmly attached to the plastic substrate. The amylase content of the cultured cells remained adequate for assay of secretory activity during culture for one week after isolation. Depending upon variations in experimental treatments, the cultured cells secreted approx. 35-65% of cellular amylase in response to a carbachol challenge. The cells were slightly responsive to long exposures to isoproterenol, and were unresponsive to nicotine, elevated extracellular K+ or substance P. Secretion induced by carbachol required extracellular Ca2+, was inhibited by atropine and occurred with a nearly linear response over a 30-min period. The Ca2+ ionophore A23187 was also a potent secretagogue for amylase secretion, producing levels of secretion equal to that induced by carbachol. The ease of preparation and the retention of amylase during primary culture suggests that the preparation will be useful in studies on muscarinic receptor-mediated control of amylase secretion.
Brooks
J C
JC
Department of Basic Sciences, Marquette University School of Dentistry, Milwaukee, WI 53233, USA.
Brooks
M
M
Piskorowski
J
J
Watson
J D
JD
D000681:Amylases / Q000378:metabolism*; D000704:Analysis of Variance; D000818:Animals; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000502:physiology; D002217:Carbachol / Q000494:pharmacology; D002478:Cells, Cultured / Q000187:drug effects / Q000648:ultrastructure; D017364:Collagenases / Q000494:pharmacology; D046508:Culture Techniques / Q000379:methods; D010450:Endopeptidases / Q000494:pharmacology; D005260:Female; D010306:Parotid Gland / Q000201:enzymology* / Q000648:ultrastructure; D011976:Receptors, Muscarinic / Q000187:drug effects; D012463:Saliva / Q000201:enzymology; D012636:Secretory Rate / Q000187:drug effects; D013268:Stimulation, Chemical; D013552:Swine / Q000502:physiology*
D011976:Receptors, Muscarinic; D000001:Calcimycin; D002217:Carbachol; D000681:Amylases; D010450:Endopeptidases; D017364:Collagenases; C023139:dispase; D002118:Calcium