Rat serum induces a differentiated phenotype in a rat parotid acinar cell line.
To establish a continuous cell line, freshly prepared rat parotid acinar cells were stably transfected with a plasmid vector containing the SV40 large T antigen. The acinar origin of these cells was confirmed by Western blotting, enzyme analysis, and morphological analysis. Transformed cells grown in 10% rat serum showed a modest reduction in cell number after 7 days and a concentration- and time-dependent increase in amylase levels approximately 16 times greater than those observed in fetal bovine serum-treated cells. Ultrastructural analysis revealed that cells grown in rat serum harbored protein-filled secretory granules localized adjacent to the endoplasmic reticulum, and punctate amylase-specific immunofluorescence distributed throughout the cytoplasm was consistent with the presence of amylase in secretory organelles. Clonal cells express tissue-specific proline-rich proteins and the four protein kinase C isozymes present in primary culture. Carbachol and isoproterenol stimulated [3H]protein secretion and isoproterenol enhanced amylase secretion from cells grown in rat serum. Moreover, norepinephrine, carbachol, and substance P produced a time- and concentration-dependent rise in cytoplasmic Ca2+. This continuous cell line of parotid acinar cells, which after treatment with rat serum retains the basic structural and functional properties of primary culture cells, will be utilized as a model system for studying long-term biological processes that regulate parotid cell function.
Department of Pharmacology and Toxicology, School of Medicine and Biomedical Sciences, State University of New York, Buffalo, New York 14214, USA.
Y
Y
Zhu
J M
JM
Aletta
J
J
Wen
X
X
Zhang
D
D
Higgins
R P
RP
Rubin
D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000096:biosynthesis / Q000235:genetics; D001769:Blood; D002417:Cattle; D002454:Cell Differentiation / Q000502:physiology*; D002461:Cell Line, Transformed; D002999:Clone Cells; D003470:Culture Media; D008297:Male; D015388:Organelles / Q000648:ultrastructure; D010306:Parotid Gland / Q000166:cytology* / Q000201:enzymology / Q000648:ultrastructure; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000096:biosynthesis; D013539:Simian virus 40 / Q000235:genetics; D014162:Transfection; D000516:alpha-Amylases / Q000096:biosynthesis
D000952:Antigens, Polyomavirus Transforming; D003470:Culture Media; D011994:Recombinant Proteins; D000516:alpha-Amylases